P < 0.05). Log-transformed levels of soluble intercellular adhesion molecule-1 (sICAM-1) were negatively correlated with relative MBR  before surgery  in macula-on, but not macula-off, RRD. Six months postoperatively, r-MBR correlated significantly with sICAM-1 levels  in macula-off RRD. ICAM-1 may play a role in RRD-induced deterioration of retinal circulation.Many studies have demonstrated that rhegmatogenous retinal detachment (RRD) leads to impaired retinal circulation. However, the involvement of inflammation in the RRD-induced worsening of retinal circulation was obscure. This retrospective observational study included 150 patients with primary RRD  who underwent 25-gauge microincision vitrectomy surgery (25G MIVS). Total retinal blood flow was represented by the mean blur rate (MBR) of the optic nerve head vessel, measured by laser speckle flowgraphy preoperatively and until 6 months postoperatively. Aqueous humor samples were obtained during surgery to determine cytokine concentrations by enzyme-linked immunosorbent assay. At 3 and 6 months postoperatively, there were no significant differences between eyes with macula-on RRD and fellow eyes. However, in macula-off RRD, MBR remained significantly lower in RRD eyes 6 months postoperatively ( Therefore, prompt repair is required to prevent photoreceptor cell damage after RRD. In general, surgical repair is by pars plana vitrectomy (PPV) or scleral buckling, both of which obtain high rates of successful reattachment with a single operation2. Recently, PPV has become the more popular procedure with the advent of microincision vitrectomy surgery (MIVS), which results in less intraoperative and postoperative pain and a shorter operation time3. However, although complete mechanical attachment can be obtained by surgery after RRD, recovery of visual function is often incomplete.Rhegmatogenous retinal detachment (RRD) is an acute retinal disorder characterized by the mechanical separation of the neurosensory retina from the retinal pigment epithelium. Since photoreceptor cells in the neurosensory retina are supported by retinal pigment epithelium cells, RRD usually induces photoreceptor apoptosis6 but was reported to recover to the levels in the fellow eyes by about 6 months postoperatively7. Because conventional methods for measuring ocular circulation are time consuming and partially invasive, it is difficult to repeat measurements to record longitudinal changes in retinal and/or choroidal circulation after surgical repair of RRD.The importance of detachment in the macular area in preoperative RRD is well known. The macula has a high density of photoreceptors, which require substantial nutrition via the retinal and choroidal circulation. Retinal circulation is impaired in eyes with RRD11. Using LSFG, Iwase et al. examined the longitudinal changes in retinal circulation measured before and after 25-gauge (25G) MIVS or scleral buckling in RRD eyes with the macula still attached (macula-on RRD)4. They demonstrated that 25G MIVS, but not scleral buckling, resulted in the recovery of retinal circulation by 6 months postoperatively4. However, to our knowledge, no study using LSFG has evaluated longitudinal changes in retinal circulation after surgical repair of RRD that involved the macula (macula-off RRD) in a relatively large cohort.Recently, laser speckle flowgraphy (LSFG) has been recognized as a useful method of measuring retinal and choroidal circulation non-invasively and without the aid of contrast medium14. However, whether intraocular cytokine levels can affect longitudinal changes in retinal circulation after RRD is unknown. Therefore, using LSFG, we compared the longitudinal changes in retinal circulation after macula-on and macula-off RRD treated with 25G MIVS. In addition, we aimed to assess whether the levels of cytokines in the aqueous humor correlated with retinal circulation in postoperative RRD eyes.In RRD eyes, inflammatory cytokines are markedly upregulated and considered a decisive factor in promoting apoptotic death of photoreceptors in the detached retinaPreoperative characteristics of the participants are shown in Table At baseline, the vessel MBR in the optic nerve head was significantly lower in RRD eyes than in fellow eyes Fig. F,G. In mTo elucidate whether impaired retinal circulation at baseline creates any harmful effects on ocular parameters 6 months postoperatively, we performed Pearson correlation coefficient analysis. BCVA and CRT at 6 months postoperatively were not correlated with r-MBR before surgery (data not shown).r =  − 0.27, P = 0.03), and 3  and 6 months after surgery .We excluded subjects with a blank cytokine measurement, leaving 92 subjects included in this additional analysis . We compared cytokine concentrations in the aqueous humor of eyes with macula-on and macula-off RRD , but not the macula-off RRD eyes . At 6 months after 25G MIVS, log-transformed sICAM-1 levels were significantly correlated with r-MBR only in the macula-off RRD eyes .To further examine the relationship between the aqueous humor levels of sICAM-1 and the time course of r-MBR in macula-on and -off RRD, we calculated the Pearson correlation coefficient before surgery and 6 months postoperatively Fig. . Before 4, 25G MIVS promoted restoration of retinal circulation in macula-on RRD. Our study demonstrated for the first time that retinal circulation in macula-off RRD remained impaired after treatment with 25G MIVS and that the aqueous humor level of sICAM-1 at baseline was negatively correlated with retinal circulation in RRD eyes postoperatively.In the present study, we observed the time course of retinal circulation in eyes with RRD by measuring vessel MBR in the optic nerve head using LSFG. Consistent with previous reports15. Our data also showed significant reductions in BCVA and CRT in macula-off RRD, indicating that neurodegenerative changes occurred at the macula. Retinal circulation in eyes with macula-off RRD did not recover to the levels observed in fellow eyes until at least 6 months after 25G MIVS. Accumulating evidence suggests that retinal circulation is severely impaired in retinal degenerative diseases such as retinitis pigmentosa and other inherited retinal diseases18. Macula-off RRD led to thinning of the macula and degeneration of photoreceptors. Therefore, the circumstances of macula-off RRD might somewhat replicate retinal degenerative disorders. Given that retinal circulation is highly regulated according to neuronal demand for oxygen and nutrients, we speculate that the persistence of deteriorated retinal circulation is associated with the severe neurodegenerative changes of macula-off RRD.The prognosis of visual function after surgical repair of RRD depends on the extent of macular involvement. Macula-off RRD gives rise to degeneration in neurons and blood vessels in the macula19. To assess the relationship between preoperative retinal circulation and postoperative alteration of neuronal retina in macula-off RRD in more detail, a future study should employ not only optical coherence tomography measurement of retinal thickness but also focal macular electroretinograms.Recovery of the neuronal retina is also a concern after successful reattachment in macula-off RRD. In the present study, the retina remained thin in macula-off RRD compared to macula-on RRD Fig. B, indica22. We found that levels of PlGF, PEDF, IL-6, IL-8, and IP-10 were more elevated in macula-off RRD than macula-on RRD. PlGF is a member of the vascular endothelial growth factor (VEGF) family and accelerates angiogenesis and inflammation23. To date, no study has examined intraocular levels of PlGF in RRD eyes. In contrast, intraocular levels of VEGF have been extensively examined in RRD and have consistently remained unaffected24. Our data further substantiate the notion that VEGF level is not altered by the RRD or its severity. PEDF exhibits anti-angiogenic effects25 and is essential for neuronal development in the retina26, and supplementation with PEDF has a significant neuroprotective effect27. Recently, Takahashi et al. reported that vitreous levels of IL-6, IL-8, and IP-10 were elevated in RRD eyes24, and IL-8 was significantly correlated with the extent of the detached area. In RRD, the role of IL-6 and IL-8 is not fully elucidated, but IP-10 was reported to be involved in photoreceptor death22. Although we did not examine the extent of retinal detachment in our patients, we speculate that a range of severity might account for differences we observed in the concentrations of these cytokines.Elevated levels of cytokines have been demonstrated in RRD eyes in the intraocular fluids, such as the vitreous and aqueous humor28, suggesting that inflammatory cytokines play a key role in the worsening of retinal circulation in eyes with central retinal vein occlusion. Our results further suggest that ICAM-1 might contribute to the deterioration of retinal circulation in cases of RRD. ICAM-1 is an adhesion molecule and promotes leukostasis in ischemic retinopathies such as retinal vein occlusion and diabetic retinopathy33, in which inflammation plays a crucial role. In the present study, sICAM-1 levels in RRD eyes were significantly correlated with IL-6 and IL-8 levels  and macula-off RRD (n = 87), which were defined respectively as the presence and absence of subfoveal fluid on preoperative clinical evaluation. To minimize variation in retinal circulation, participants were excluded if they had diabetes or ocular abnormalities such as glaucoma, uveitis, or a history of ocular surgery. Informed consent was obtained from all participants. We acquired the decimal best-corrected visual acuity (BCVA), intraocular pressure (IOP), and central retinal thickness (CRT) using spectral-domain optical coherence tomography . At every visit, blood pressure (BP) was monitored. For analyzing variables affecting retinal circulation, mean arterial pressure (MAP) was calculated as MAP = diastolic BP + (systolic BP − diastolic BP)/3. Ocular perfusion pressure (OPP) was calculated as OPP = 2/3 MAP − IOP. Decimal BCVA was converted to the logarithm of the minimal angle of resolution (logMAR) scale for statistical analysis.We reviewed the records of 150 patients with RRD who underwent 25G MIVS from September 2014 to June 2018 at Hachioji Medical Center, Tokyo Medical University. Patients were categorized into macula-on RRD (6) was used as the gas tamponade.All procedures were performed under local anesthesia via sub-Tenon injection using 2% xylocaine. Immediately after anesthetic injection, we collected the aqueous humor with a 30G needle and stored it at − 80 °C until use. Cataract surgery was performed with phacoemulsification and aspiration (PEA) if necessary. All cases of PEA were followed by intraocular lens implantation. A 25G MIVS  wide-angle viewing system  was used for PPV, during which the vitreous base was shaved circumferentially as much as possible with partial scleral depression. After fluid–air exchange, conventional laser photocoagulation was performed. Filtered room air or 20% hexafluoride sulfate  and a charge-coupled device were used to obtain an image of a speckle pattern that was generated by scattering light from erythrocytes. The mean blur rate (MBR), which reflects the relative velocity of blood flow36, was calculated from the variation of the blurring in the speckle pattern and expressed in arbitrary units (AU). We analyzed the average MBR of the vessels on the optic nerve head using LSFG Analyzer software 4. To normalize the MBR on the optic nerve head in RRD eyes, we also calculated relative MBR (r-MBR) by dividing the MBR value of the RRD eye by that of the fellow eye.Retinal circulation was measured using LSFG-NAVI  as detailed previously37. Briefly, 100 μL of aqueous humor per subject was applied to MILLEPLEX MAP® Human Cytokine/Chemokine Magnetic Bead Panel-Immunology Multiplex Assay . The procedures were performed according to the manufacturer’s instructions. In the analyses of the relationship between cytokine levels and r-MBR, cytokine levels were log-transformed28.Cytokine concentrations in the aqueous humor were measured by enzyme-linked immunosorbent assay, as previously reportedP < 0.05 was considered significant.Data are expressed as the mean ± standard deviation. Statistical comparisons of the parameters between the groups were performed using the Mann–Whitney U-test. Two-way repeated-measures analysis of variance and post-hoc Bonferroni multiple-range test were used to identify significant differences in the parameters between macula-on and -off RRD at baseline and at postoperative visits at 1, 3, and 6 months. Pearson correlation coefficient was calculated to determine whether elevated cytokine levels affect postoperative retinal circulation. Supplementary Information 1.Supplementary Information 2.


Zishen Yutai Pills (ZSYTP) is a prescription based on traditional Chinese medicine used to treat kidney-deficient pattern in traditional Chinese medicine. It is also widely used clinically for the treatment of polycystic ovary syndrome (PCOS) with positive results. This study aims to explore the potential pharmacological mechanism of ZSYTP for the treatment of PCOS by a network pharmacology approach.  Compounds were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine and TCM Database@ Taiwan, and the corresponding targets were retrieved from PubChem, Swiss Target Prediction, STITCH, and DrugBank. Meanwhile, PCOS targets were retrieved from the GeneCards database, the Online Mendelian Inheritance in Man database, National Center for Biotechnology Information Database, and DrugBank. Subsequently, multiple network construction and gene enrichment analyses were conducted with Cytoscape 3.8.2 software. Based on the previous results in the study, molecular docking simulations were done.  205 active compounds and 478 ZSYTP target genes were obtained after screening by ADME consideration. 1881 disease-related targets were obtained after removing duplicates. 148 intersection target genes between drug and disease targets were isolated. Gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes analysis highlighted multiple gene functions and different signaling pathways to treat PCOS. Further molecular docking demonstrated the practicality of in vivo action of ZSYTP to a certain extent.  It is possible that the pharmacological effect of ZSYTP on PCOS is linked to the hypoxia-inducible factor 1 (HIF-1) signaling pathway, improving insulin resistance, the variation on gene expression such as RNA splicing, and regulation of mRNA metabolic process. This study paves the way for further research investigating its mechanisms. Polycystic ovary syndrome (PCOS) is a common gynecological disorder affecting 5%–15% of women of reproductive age from all over the world .At present, due to the relatively unknown etiology of PCOS, there is no effective treatment protocol. Symptomatic treatment is mainly used in clinical practice, and long-term health management is needed. Traditional Chinese medicine (TCM) has long been utilized to treat infertility for centuries and has been proven effective in managing the symptoms of PCOS patients and promoting ovulation. In accordance with TCM theory, PCOS patients who have obesity or insulin resistance  can be wArtemisia carvifolia led to the discovery of artemisinin and subsequently became crucial in the fight against malaria [In recent years, increasing research has been done on traditional or patented TCM formulae, and it can be a gift for drug discovery . For ins malaria .A network pharmacology approach was chosen to uncover ZSYTP's pharmacological mechanism on PCOS, aiming to provide a protocol for studying patented formulae and a specific disease. This study also would provide the data and direction for further clinical trials to develop, improving the efficacy of future drug development studies.https://old.tcmsp-e.com/tcmsp.php), a unique systemic pharmacology platform devised for Chinese herbal medicines.Chemical compounds from 15 herbs in ZSYTP were obtained from the Traditional Chinese Medicine System Pharmacology Database  , which is a bioinformatics analysis tool specially designed for the research of the molecular mechanism of TCM and TCM Database@ Taiwan [http://tcm.cmu.edu.tw/zh-tw/), the world's largest noncommercial TCM database.The relationships between drugs, targets, and diseases were obtained from a Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine   which could ensure higher efficiency to the active compounds selected.http://pubchem.ncbi.nlm.nih.gov/) to corroborate the uniqueness of these molecules in the database we built.We collated IUPAC International Chemical Identifier (InChi) and Canonical SMILES of each 205 chemical compounds of ZSYTP from PubChem database  (http://http://stitch.embl.de) and Swiss Target Prediction [http://www.swisstargetprediction.ch/index.php). PubChem database was utilized to provide drug-target identification. Finally, we found information on known drugs and their corresponding target genes in DrugBank [https://go.drugbank.com/), which is a pharmaceutical knowledge base.To predict interactions between chemicals and proteins, we input all molecular information of the 205 active compounds into STITCH  (http://ediction  (http://DrugBank (https://http://www.genecards.org/), the Online Mendelian Inheritance in Man database [http://www.omim.org/), National Center for Biotechnology Information database [https://www.ncbi.nlm.nih.gov/), and DrugBank database (http://www.drugbank.ca/). GeneCards, OMIM, and NCBI database provide comprehensive data on predicted and annotated human genotypes, while DurgBank offers data on the disease and the drugs available to it. Due to the nonstandard naming of the proteins collated from the 3 databases, we used the UniProt database [http://www.uniprot.org/) to match the standard gene names.“Polycystic Ovary Syndrome” is the keyword used to retrieve relevant action targets. To obtain more comprehensive disease-related targets, 3 online databases were utilized, including the GeneCards database   ZSYTP active compound-potential target network (C-T network), (2) PPI network of PCOS disease targets, (3) PPI network of ZSYTP drug targets, and (4) PPI network of interaction between PCOS disease and drug targets.https://bioinfogp.cnb.csic.es/tools/venny/), an online program, was utilized to visualize and isolate overlapping targets between the active ZSYTP compounds and those related to PCOS. The data were imported into Cytoscape 3.8.2 software to construct the active compound-potential target network (C-T network). The nodes represent the target genes and compounds, and the line edges represent the correlations between them. The size and transparency of the nodes were set to reflect the degree value. The active compounds and target with the highest degrees were selected for future molecular docking analysis.Venny 2.1 , and the small-molecule compounds which are considered as part of the first five high degree value coming from C-T network were retrieved via the PubChem database and TCMSP. Expulsion of water and ligand from macromolecular protein downloaded from PDB was performed using PyMOL 2.4 software [Molecular docking simulations were utilized to verify the binding of the target and the corresponding compound. Data about the construction of the macromolecular protein target receptors, including the first eight high degree values which come from the C-T network, were acquired via the RCSB PDB database  , 188 from Lycii Fructus (GQZ), 174 from Morindae officinalis Radix (BJT), 165 from Amomum aurantiacum (SR), 135 from Folium Artemisiae Argyi (AY), 134 from Codonopsis Radix (DS), 119 from Eucommiae Cortex (DZ), 76 from Rehmanniae Radix Praeparata (SDH), 55 from Atractylodes Macrocephala Koidz (BZ), 46 from Herba Taxilli (SJS), 30 from Dipsaci Radix (XD), 29 from Cuscutae Semen (TSZ), 17 from Fallopia multiflora (HSW), 4 from Colla Corii Asini (EJ), and 2 from Cornu Cervi Degelatinum (LJS).1364 chemical compounds in ZSYTP were found in this process; there are 190 from Upon application of ADME screening thresholds of OB ≥ 30% and DL ≥ 0.18%, 187 chemical compounds were obtained: 22 from RS, 45 from GQZ, 20 from BJT, 10 from SR, 9 from AY, 21 from DS, 28 from DZ, 2 from SDH, 9 from BZ, 2 from SJS, 6 from XD, 11 from TSZ, and 2 from HSW. LJS and EJ were not found in any online TCM databases; however, further 18 compounds with lower OB or DL values were consolidated as they hold extensive pharmacological activities, 10 from HSW, 4 from EJ, and 2 from LJS. These 205 compounds were considered as active compounds with good biological activity.205 active compounds were based on molecular similarity to predict the target genes, which was conducted by importing each unique molecular data into STITCH, Swiss Target Prediction, PubChem, and DrugBank. 478 target genes of every herb of ZSYTP were acquired after eliminating duplication.A total of 1881 PCOS disease-related targets were obtained upon removing duplicates. Further comparison was made between the 478 potential target genes of ZSYTP and 1881 PCOS disease targets; 148 intersection potential target genes that act on PCOS  were subC-T network illustrates the relationship among each herb, its corresponding active compounds, and targets. These were mapped out by the Cytoscape 3.8.2 software. The nodes describe herbs, active compounds, and target genes, and the edges indicate the relevance among them. A C-T network can provide further insights into the study of ZSYTP's therapeutic mechanism on PCOS.The constructed C-T network of 39 active compounds includes 200 nodes and 540 edges depicted in Compounds and targets with the highest degree values were utilized to do molecular docking verification. The degree values are listed in PPI network of the intersectional potential target genes between disease and drug targets was constructed with Cytoscape version 3.8.2 BisoGenet plug-in.8937 nodes and 200243 edges were established. Based on a previous study, topological feature analysis targets were selected with parameters above twice the median value. Upon the first screening process, a network of 2469 nodes and 101184 edges was derived. The second selection criteria were based on “Betweenness,” “Closeness,” “LAC,” and “Neighborhood Connectivity”; 473 nodes and 10130 edges were identified. The topological screening process is presented in p values less than 0.05 were defined as statistically significant. The visualization in this part is shown in Cytoscape 3.8.2 software ClueGO plug-in was utilized to explore the active process of ZSYTP on PCOS by the 473 relationship genes from the PPI network, and p-values less than 0.05 as statistically significant. 540 processes were obtained. The top 20 biological processes with ascending p-values were selected for further analysis.The enrichment analysis of BP was completed with the Cytoscape ClueGO plug-in for visualization , with p-The biological processes selected were mainly involved in RNA splicing, cellular macromolecule metabolic process, regulation of mRNA metabolic process, cellular protein metabolic process, cellular response to stress, intracellular transport, regulation of the nucleobase-containing compound metabolic process, and regulation of nitrogen compound metabolic process.p values were selected for further analysis and visualized in p value.107 terms were retrieved in gene ontology analysis of MF. Twenty molecular functions with the ascending p values were selected for further analysis and visualized in 114 components were identified in gene ontology analysis of CC, including focal adhesion, cytosolic large ribosomal subunit, spliceosomal complex, cytosolic small ribosomal subunit, vesicle, extracellular space, catalytic Step 2 spliceosome, nucleolus, nuclear chromosome, chromosome, anchoring junction, chromosomal region, nuclear body, nuclear speck, polysome, and nuclear periphery. Twenty cellular components with ascending Further KEGG analysis highlighted 16 pathways of ZSYTP potential pharmacological mechanisms on PCOS. All pathways are presented in The lowest binding energy of molecular docking of potential targets and their appointed compounds is shown in The molecular docking simulations of GSK3B helenalin, F2 beta-sitosterol, F2 helenalin, DPP4 helenalin, CYP1B1 beta-sitosterol, and CYP1B1-helenalin are shown in Figures 7These figures show the residues and hydrogen bonds that the ligand binds to the protein. The residues are indicated in bluish violet, while the hydrogen bonds are dotted in yellow.Lycii Fructus, Cuscutae Semen, Eucommiae Cortex, Artemisiae argyi, Panax ginseng, Morindae officinalis Radix, and Amomum aurantiacum and helenalin in Eucommiae Cortex among the disease targets is shown to be stable and feasible. Therefore, these may warrant more research for further validation of the therapeutic effects of ZSYTP.Visualization of beta-sitosterol in ZSYTP was first used to prevent recurrent spontaneous and threatened abortion. In line with TCM's theory of treating different diseases with the same therapeutic principle and on the basis that the kidney in TCM's theory is closely related to reproduction, ZSYTP is also used to treat irregular menstruation and infertility in clinical practice, such as PCOS –30.It has been found that ZSYTP can improve the clinical symptoms of PCOS patients . TherefoIn order to further support the existing literature and provide insights and possible hypotheses for the pharmacological mechanism of ZSYTP in the treatment of PCOS, we conducted extensive literature research on the active compounds as highlighted by the PPI networks and 16 pathways as indicated by KEGG pathway enrichment analysis.As a tonifying kidney TCM formula, ZSYTP is found to have therapeutic effects in the treatment of POI and PCOS . HoweverOur team conducted a network pharmacology study on the effect of ZSYTP on POI. One hypothesis put forward by our previous study was that ZSYTP upregulates the hypoxia-inducible factor-1 (HIF1) signaling pathway , therebyα, which is regulated by the low oxygen concentration of the environment [HIF-1 is a transcription factor, and its activity is mainly determined by HIF-1ironment . Human oironment  and a loironment . Developironment .One study showed that polycystic ovaries were found in rats when submitted into hypobaric hypoxia. Weight of the ovaries increased under a hypoxic environment, while synthesis and secretion of E2 were negligible .α protein expression in PCOS women were significantly lower than those of the control group, indicating HIF-1α′s possible involvement in the molecular mechanisms of endometrial dysfunction in PCOS women, affecting fertility [Multiple animal experimental studies suggest that HIF-1 promotes follicular development and interacts with FSH receptors on granulosa cells to regulate E2 secretion . One othertility .The balance between HIF-1 expression and oxygen concentration of the environment is therefore crucial for normal follicle development and subsequent ovulation to take place.α in a manner similar to hypoxia and stabilizes HIF-1α [Quercetin is an active compound in ZSYTP with the highest degree, as indicated in the Compound-Potential Target Network . It has s HIF-1α . Quercets HIF-1α .It is possible that the active compounds in ZSYTP like quercetin activate the HIF-1 transcription factor, promoting follicular development in PCOS patients and thereby reducing follicular atresia and regulating ovulation.PCOS women have an increased risk of insulin resistance (IR), while multiple studies have shown that IR plays an important role in the occurrence and development of PCOS as well . Due to When PI3K is activated, it affects the intracellular glucose transporter (GLUT) to transport glucose across the membrane. Studies have shown that GLUT4 expression in the PCOS uterus is significantly reduced, thus affecting glucose metabolism in the body. Quercetin, the active ingredient in ZSYTP, can increase the expression of this gene . In addiNormally, all female follicles develop through a series of processes such as recruitment, selection, dominance, and ovulation, in which physiological concentrations of insulin levels promote granulosa cell proliferation, and glucose metabolism in granulosa cells provides energy for oocyte development, thereby regulating follicular development. In addition, the insulin signaling pathway affects ovarian physiological function, oocyte developmental quality, and the ovulation process .In the PPI network, through the analysis of the active components and targets of ZSYTP, it is predicted that the main active components and targets of ZSYTP have a good potential to improve IR in PCOS. Not only quercetin but also luteolin and emodin have a multidirectional regulation effect on the etiology of PCOS that has been explored .Luteolin and emodin can improve the environment of high insulin levels by promoting PI3K/AKT pathway , 51 and Dipeptidyl peptidase-4 (DPP4) is one of the main potential target genes of ZSYTP and PCOS, and studies have shown that it plays a key role in the regulation of insulin secretion. DPP4 serum activity is elevated in patients with polycystic ovary syndrome, DPP4 serum levels are closely related to anti-Müllerian hormone (AMH) serum levels , and thePatients with PCOS often have symptoms of ovulatory disturbance or anovulation, which may be related to their IR, and our results from molecular docking suggest that ZSYTP may improve IR associated with PCOS, thus directly or indirectly improving ovarian function.Gene expression in eukaryotes is performed by RNA polymerase, spliceosome, and ribosome, respectively , and KEGThe pathogenesis of PCOS still remains undefined; therefore, there is no effective cure for this disease. Although most fertility problems in PCOS patients can be resolved through symptomatic treatment, PCOS still severely affects women's long-term health.Hence, this paper aims to investigate the possible etiology of PCOS and explore the pharmacological mechanism between ZSYTP and PCOS. We extracted 205 active compounds from ZSYTP and simultaneously retrieved 1881 disease target genes on multiple databases. 148 intersection target genes were derived by intersecting the databases of disease and drug-target genes.Subsequently, our team constructed C-T network and multiple PPI networks and performed the GO and KEGG analysis. This study hypothesizes that ZSYTP may alleviate IR to treat PCOS. GO and KEGG analysis also highlighted that the HIF-1 signaling pathway and gene expression could be the pathological mechanisms of PCOS.Additionally, the result of this research shows that the same compounds in ZSYTP could regulate multiple targets; meanwhile, the same targets could intervene in multiple biological processes and pathways.It reflects the characteristics of the combined action of multipathway and multitarget of ZSYTP. The results suggested that the whole network can be regulated by regulating single or multiple targets in the network, which provides a scientific basis and new insights for the clinical application of ZSYTP in the treatment of PCOS and simultaneously provides a new direction for further exploring the potential mechanism of ZSYTP on PCOS.This study is mainly based on bioinformatics and massive data calculation results, which will be further screened and verified by animal or cell experiments to clarify the main regulatory targets of ZSYTP, thus promoting the development of modern pharmacology of TCM.


This study aims to clarify the potential mechanism of modified Bu-Shen-Huo-Xue decoction (MBSHXD) in treating intervertebral disc degeneration (IDD) with methods of network pharmacology and molecular docking.  An MBSHXD and IDD-related common target gene set was established through TCMSP, UniProt, and two disease gene databases. GO and KEGG enrichment analysis and protein-protein interaction (PPI) networks were performed through the R platform and STRING to discover the potential mechanism. Molecular docking between the active ingredients and the core genes is used to calculate the binding energy.  A total of 147 active ingredients and 79 common genes  were identified. The results of GO and KEGG enrichment analysis showed that MBSHXD plays an essential role in regulating inflammation and oxidative stress. The meaningful pathways are the AGE-RAGE signaling pathway in diabetic complications, the IL-17 signaling pathway, the TNF signaling pathway, the PI3K-Akt signaling pathway, the MAPK signaling pathway, and apoptosis. In addition, the PPI network and molecular docking further demonstrated the roles that nine bioactive ingredients of MBSHXD play in IDD treatment through their interference with core target proteins.  This study reveals that MBSHXD has the characteristics of a “multi-component, multi-target, and multi-pathway” in the treatment of IDD by regulating inflammation and oxidative stress, and network pharmacology may provide a feasible method to verify the molecular mechanism of MBSHXD for IDD by combining with molecular docking. As a common symptom, low back pain is the leading cause of years living with disability . It is rRadix Rehmanniae Preparata, Cortex Eucommiae, Radix Aconiti Lateralis Preparata, Fructus Lycii, Fructus Corni, Semen Persicae, Flos Carthami, Rhizoma Dioscoreae, and Radix Glycyrrhizae. According to TCM, MBSHXD has the effects of nourishing the kidney, promoting blood circulation, and strengthening the bones. Recent studies have shown that MBSHXD may delay the progression of degenerative diseases of the musculoskeletal system, including osteoarthritis and IDD [In recent years, traditional Chinese medicine (TCM), as an essential branch of complementary and alternative medicine, has received increasing attention for its role in various diseases –5. Modif and IDD , 7. Zhu Network pharmacology, an emerging and interdisciplinary research approach, is based on traditional pharmacology, bioinformatics, chemoinformatics, and network biology , 9. The https://tsmspw.com) [http://www.drugbank.ca) [https://www.uniprot.org/) [We got the active ingredients of each herb in MBSHXD through the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database (spw.com) . The orabank.ca) .Then, weot.org/)  to obtaihttps://www.genecards.org) [https://omim.org, update November 25, 2020) [https://www.uniprot.org/) [IDD-related targets were obtained through retrieving GeneCards (rds.org)  and Onli5, 2020) . The OMIot.org/) .A common gene set between MBSHXD targets and IDD-related genes was built by the Venn diagram. According to a common gene set, we identified the active components corresponding to each gene and constructed an “active ingredient-common target gene network” using Cytoscape 3.7.2 software . In addiThe biological functions of genes were revealed in three different aspects, including biological processes (BPs), cell components (CCs), and molecular functions (MFs), by gene ontology (GO) enrichment analysis. The Kyoto Encyclopedia of Genes and Genomes (KEGG) is a database resource for understanding high-level functions and utilities of the biological system, such as cells, organisms, and ecosystems, from molecular-level information. We used the clusterProfiler package of the R platform for GO and KEGG functional enrichment analysis .https://www.string-db.org/) [Homo sapiens” and a confidence score >0.4. In the same way, the PPI network of IDD targets was also established. The PPI network of the common target was imported into the Cytoscape software to make it visible. A cytoHubba plugin [The common genes of MBSHXD targets and IDD-related genes were used to build a PPI network through the STRING database (db.org/) . In thisa plugin  was useda plugin . The para plugin .https://www.rcsb.org/) and PubChem database (https://pubchem.ncbi.nlm.nih.gov/), respectively. The energy of the molecular ligand was optimized using the MM2 calculation method and derived using the ChemBio 3D software. The receptor protein was dehydrated and small-molecule ligand was removed with PyMOL 2.4.0 software. The AutoDock tool performed hydrogenation of proteins and active ingredients. Finally, AutoDock Vina was used to calculate the binding energy between the receptor protein and active components [The 3D structures of the proteins encoded by the core genes and the 2D structures of active ingredients were downloaded from the RSCB PDB database (mponents .Radix Rehmanniae Preparate (Shudihuang), Cortex Eucommiae (Duzhong), Radix Aconiti Lateralis Preparate (Fuzi), Fructus Lycii (Gouqizi), Fructus Corni (Shanzhuyu), Semen Persicae (Taoren), Flos Carthami (Honghua), Rhizoma Dioscoreae (Shangyao), and Radix Glycyrrhiza (Ganzao). A total of 244 active ingredients of MBSHXD were obtained through searching TCMSP  organization, extracellular structure organization, and extrinsic apoptotic signaling pathways. The molecular functions of IDD would be related to cytokine activity, cytokine receptor binding, and signaling receptor activator activity. The cellular components associated with IDD might activate the collagen-containing ECM, endoplasmic reticulum lumen, and external side of the plasma membrane . MeanwhiTo further explore the interactions between common target genes and the mechanisms by which MBSHXD may treat IDD, GO enrichment analysis and KEGG pathway enrichment were performed using the R platform. We acquired a total of 2114 entries about biological processes. The top 10 are shown in Firstly, we inputted 79 common genes into the STRING to construct the PPI network . To furtAfter comparing and analyzing the KEGG enrichment of the disease targets and common targets of MBSHXD and IDD, we found 138 overlapping KEGG pathways  of MBSHXD against IDD by the MCC algorithm. These core target proteins are involved in multiple signaling pathways such as inflammation, immunity, metabolism, and proliferation-related signaling pathways. Studies have confirmed that the process of IDD can be delayed by inhibiting the TNF-ing IL-6 , 45. It ing IL-6 . Akt1 ising IL-6 . Moreoveing IL-6 .κB axis [According to our “disease-core genes–active ingredients–herbs” network, nine bioactive compounds between the 10 core genes and the corresponding active compounds in MBSHXD were identified. Furthermore, three hub ingredients, quercetin, luteolin, and kaempferol, were selected, and they targeted the largest number of core genes. In vitro experiments showed that quercetin ameliorated the progression of IDD by suppressing the expression of senescence associated secreted phenotype factors and improving the senescence phenotype of NP cells via the Nrf2/NF-κB axis ; luteoliκB axis ; kaempfeκB axis . These cα stimulation [Based on the ten core genes and their corresponding core compounds, molecular docking was performed to demonstrate the binding energy between proteins and small-molecule compounds, which all showed good binding affinity. The abovementioned molecular docking results were presented in the form of a heat map by using R software. In addition, the software package ClusterProfiler was used for enrichment analysis of GO and KEGG pathways. The results of two enrichment analyses of GO and KEGG were compared with the IDD-related pathways searched in PubMed. We speculated that the therapeutic effect of MBSHXD in IDD may be related to its anti-inflammatory and antioxidative stress effects, which inhibit apoptosis and senescence of NP cells. Indeed, inflammation, ECM degradation, and senescence of NP cells are thought to be the main pathogenic mechanisms of IDD , 53. Themulation . Illien–mulation  suggesteHowever, our study also has some limitations. For example, the results of this study lacked validation by in vitro experiments, while further external validation in animals should be performed. In addition, the database we selected may not be comprehensive enough, and the active ingredients and related genes screened through the database may have been missed.In conclusion, we reveal that MBSHXD has the characteristics of “multi-component, multi-target, and multi-pathway” in IDD treatment based on network pharmacology and molecular docking. MBSHXD retards the progression of IDD by regulating the antioxidant stress and inflammatory response, which inhibits apoptosis and senescence of NP cells. The primary mechanism is related to 9 core bioactive components, 10 core genes, and 9 related pathways. This study may inspire novel treatment strategies for IDD and inform future research.


Saussurea involucrata (SAIN), also known as Snow lotus (SI), is mainly distributed in high-altitude areas such as Tibet and Xinjiang in China. To identify novel targets for the prevention or treatment of lung adenocarcinoma and lung squamous cell carcinoma (LUAD&LUSC), and to facilitate better alternative new drug discovery as well as clinical application services, the therapeutic effects of SAIN on LUAD&LUSC were evaluated by gene differential analysis of clinical samples, compound target molecular docking, and GROMACS molecular dynamics simulation.Through data screening, alignment, analysis, and validation it was confirmed that three of the major active ingredients in SAIN, namely quercetin (Q), luteolin (L), and kaempferol (K), mainly act on six protein targets, which mainly regulate signaling pathways in cancer, transcriptional misregulation in cancer, EGFR tyrosine kinase inhibitor resistance, adherens junction, IL-17 signaling pathway, melanoma, and non-small cell lung cancer. In addition, microRNAs in cancer exert preventive or therapeutic effects on LUAD&LUSC. Molecular dynamics (MD) simulations of Q, L, or K in complex with EGFR, MET, MMP1, or MMP3 revealed the presence of Q in a very stable tertiary structure in the human body.There are three active compounds of Q, L, and K in SAIN, which play a role in the treatment and prevention of non-small cell lung cancer (NSCLC) by directly or indirectly regulating the expression of genes such as MMP1, MMP3, and EGFR. In 2021, among many types of cancers, lung cancer is still among the tough, not only because its mortality is as high as 22% in both men and women [According to the microscopic morphological characteristics of lung cancer cells, lung cancer can be divided into two types: small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) . In prevCurrently, the world's targeted therapeutics for NSCLC have issues, including high R&D cost, a relatively expensive market price, and a long administration cycle. Therefore, these restrictions do not allow every NSCLC patient to use these drugs normally. Lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) are the two most common types of NSCLC, among which, LUAD accounts for far more cases than LUSC, and has become the major type of targeted therapy for NSCLC.Saussurea involucrata (SAIN) is mainly distributed in high altitude areas such as Xinjiang and Tibet in China. Research on the pharmacological effects of SAIN started in 1980  and has Network pharmacology is a new discipline that is based on the theory of systems biology to select specific signal nodes for multi-target drug molecule design . Networkhttps://tcmsp-e.com/) [Traditional Chinese Medicine Systems Pharmacology Database (TCMSP) (-e.com/)  is one oAmong them, the DL value for this system is calculated following Eq. , and to https://www.drugbank.ca)database [In the equation, x represents the descriptive index of all ingredients in SAIN, and y represents the descriptive index from the drug bank  database [The structural formula of the active ingredients of SAIN obtained above Fig. c was plodatabase  for targdatabase  was usedhttps://www.ncbi.nlm.nih.gov/geo/) database [http://gepia.cancer-pku.cn/) database [To further screen the active ingredients and their targets that were initially screened, a large number of clinical case samples of LUAD&LUSC were obtained from the GEO (database . Differedatabase . In addihttps://david.ncifcrf.gov/) database [To evaluate the mutual influence among the targets screened above, a protein interaction network and an active ingredient target pathway network were constructed. The biological processes and pathways that each target participates in in vivo were explored, and data of target genes were retrieved through the David  database [http://cello.life.nctu.edu.tw/) database [http://busca.biocomp.unibo.it/) database [In recent years, subcellular localization prediction has been a more popular method of subcellular localization. Existing data to create a database of the sequence relationships of the target sequences and subcellular structures of various genes and their regulation can accurately predict the location of the target protein on various organelles and cell membranes, and has multiple advantages. Currently used subcellular localization prediction tools include (1) the PSORT II  Thedatabase  uses thedatabase ; (3) Thedatabase  employs database , 25.Combined with the analysis of biological processes, using PSORT II, CELLO, and BUSCA databases for subcellular location prediction, respectively, and to compare the prediction results of the three tools, the intersection part was selected and the main location of the target protein in the cell was obtained.In clinical studies, to evaluate the efficacy of a drug and know the survival data, such as the survival time of patients after surgery, our analysis of these survival data is called survival analysis.After a series of screening and analysis of the ingredients and action targets of SAIN, prognostic overall survival analysis was performed on the gene IDs of the targets of the final active ingredient actions in SAIN, so as to explore the length of time that patients can survive when drugs act on these targets over time.Based on molecular docking studies, differential gene analysis, and the prediction of subcellular localization and survival analysis, the results were validated with molecular dynamics (MD) simulations, a means to computationally model the motion of a small molecule in an organismal environment .MD simulations were performed with GROMACS software , thereby12 ps). In addition, the root mean square deviation (RMSD) of the MD simulation results analysis and visualization were performed using a GROMACS inbuilt program with the RMSD formula as follows (3).After state equilibration of all environments, we performed 50 ns MD simulations of the ingredient target complex system using epidermal growth factor receptor (EGFR) with L, Q, matrix metalloproteinase 3 (MMP3) with Q, hepatocyte growth factor receptor (MET) with L, and matrix metalloproteinase 1 (MMP1) with L, Q, and K, in which stored conformations were calculated every 10 ps , and N refers to the number of atoms.In this equation, RIn this study, a total of 55 known active ingredients in SAIN were collected from the TCMSP database, including alkaloids, lipids, flavonoids, and flavonoids. Subsequently, six potent drug-resistant ingredients from 55 compounds were screened under the following screening conditions of OB ≥ 30%&DL ≥ 0.18&logPo/w ≤ 5. Details of the screening process of these six components, Flazin (F), Q, K, L, Alloisoimperatorin (A), and Hispidulin (H), are shown in Table By merging 1023 targets screened from the STP database and 764 LUAD and LUSC targets screened from the TCMSP database, a total of 9 targets with six ingredient roles was obtained. The nine protein targets interacted with the ingredients of six SAIN, and an ingredient target network was constructed using Cytoscape, resulting in an ingredient target network characterized by 15 nodes and 24 edges , 90 kDa heat shock protein ΑA1 (Hsp90AA1), MMP1, and EGFR.https://portal.gdc.cancer.gov/) database [To ensure the accuracy of the experimental data, further screening for ingredients and targets was performed. In this study, data from a total of 22,589 LUAD and 22,697 LUSC related disease genes was obtained through the GEO database and R 4.0.4, TCGA data through the GDC  of these data and the patient samples to obtain the gene expression matrix of two groups, LUAD & Normal and LUSC & Normal, corresponding to the nine targets. In this study, 15 samples from each of the two groups within the two cohorts were selected, including 30 clinical samples from each of the LUSC & control group in LUAD and LUSC, and the gene differential heat maps of LUAD Fig. B and LUS2FC value, and Q value of 1 and 0.01 were selected by the difference method [The results of gene difference analysis showed that the difference of Hsp90AA1 between LUAD and LUSC was least obvious, therefore, this was discarded. The gene IDs of the remaining eight targets were TPM analyzed by GEPIA database, input gene ID, one-way analysis of variance (ANOVA) with Loge method , and theWhen combining the correspondence between components and targets, and the results of gene difference analysis and TPM analysis, the data showed that the expression and changes of MMP2 and PTGS2 in group T were consistent with that in group N, therefore, they were discarded.The SAIN components with major effects on LUAD and LUSC were Q, l, and K, and the genes with major effects on LUAD and LUSC were MMP1, MMP3, EGFR, MET, NAD (P) H: quinone oxidoreductase 1 (NQO1), and myeloperoxidase (MPO).For the analysis of six genes, we concluded that the expression of two genes, EGFR and MMP3, were much more effective in LUSC than in LUAD, whereas MET had the opposite effect. Although the other three genes  showed consistent expression in the occurrence and development of LUAD and LUSC, MPO was of interest as this gene showed full-stage high expression in cases in the N group and low expression in cases in the T group. Therefore, we speculated that overexpression of MPO in tissues might be effective in inhibiting the development of LUAD and LUSC.Based on the above-mentioned analysis, three targets, included Hsp90AA1, PTGS2, and MMP2 were excluded, and previous studies have shown that Hsp90AA1 and the isotype Asp90AB1 may be required for the treatment of two subtypes of breast cancer (MCF-7 and BT-474) . PTGS2 iThe remaining six targets  were subjected to undergo BP analysis Fig. a, KEGG eTo achieve credibility and statistical significance of the data, we performed GO (BP) analysis and KEGG enrichment analysis at FDR ≤ 0.05. Referring to the results of KEGG enrichment analysis, and to make it easy to understand, we used Cytoscape to associate the three components, six targets, and 20 pathways to draw a component target pathway diagram  network diagram, targets were sorted into EGFR, MET, MMP1, MMP3, NQO1, and MPO based on the number of regulatory pathways and corresponding components, and pathways were ranked into pathways in cancer, transcriptional misregulation in cancer, hepatocellular carcinoma, bladder cancer, and EGFR tyrosine kinase inhibitor resistance. According to the degree of correlation adherens junction, IL-17 signaling pathway, melanoma, non-small cell lung cancer, relaxin signaling pathway and microRNAs in cancer, etc. However, although these pathways contribute to both LUAD and LUSC, the strength of the contribution is still indistinguishable, and therefore, further analysis and validation  are needed.After a series of analysis, screening, and validation efforts as mentioned above, we identified the target sites where the three components  act separately, where the three components can act on multiple targets, where, Q can act on MMP1, MMP3, EGFR, MPO, and NQO1; L can act on MMP1, EGFR, and MET; and K only acts on MMP1.https://www.rcsb.org/) database [After obtaining this information, the structures of the 6 components were plotted using ChemDraw ultra 12.0 software , saved adatabase . PDB fildatabase  , MET , MMP1 , MMP3 , MPO , and NQO1 . The larger the value in this coordinate, the greater the affinity for docking, and the greater the affinity (interactions existing between ligand and receptor), thereby demonstrating that the tighter the binding of the component to the target, the more effective the component is. However, to guarantee the availability and reference value of the data, we set the coordinates  according to the structure size of the ingredients. Therefore, the coordinate values that were used in this study were appropriate for the ingredients. The best value was − 10.1 kcal/mol and the worst was − 7.9 kcal/mol.Based on the molecular docking results, items with affinities ranging from values  were used, and the 3 SAIN components and 6 targets for which detailed results of molecular docking are given in Table To probe the specific location in the cell of the six proteins that were screened, the PSORT II, CELLO, and BUSCA databases were used for subcellular localization predictions and predictions from three databases PSORT II Tables , 4 and 5Subcellular localization prediction showed that MMP1, MMP3, NQO1, EGFR, MET, and MPO were found in the cytoplasm, and among them, MMP1, MMP3, NQO1, and MET were also found in the mitochondria, in addition to MMP3. The other five proteins were all found in the nucleus.The purpose of survival analysis is to provide a patient with the option to continue treatment before undergoing surgery or other treatments, and is derived from the analysis of genetic data associated with the disease when the patient can survive after treatment with the corresponding target .Survival analysis of these six targets showed that all six genes showed statistical significance in the regulation of LUAD (P < 0.05), while only EGFR and MMP3 were statistically significant in the regulation of LUSC (P < 0.05), which was consistent with our previous TPM analysis Fig. .Fig. 8ReRMSD data are an important basis for measuring the stability of the system and can be used to measure the stability of the tertiary structure of a protein after incorporating small molecules . To makeEGFR, NSCLC targets have been used in clinic, and the RMSD change trend and starting value in the time period of 0–50 ns are similar to those of the MMP3 protein. In addition, EGFR and MMP3 exhibit a very stable conformation (average RMSD < 0.15) after forming a complex under the action of L or Q. Similarly, for clinical use with the target MET, its complex formed under the action of L was relatively stable, but the MMP1 protein had a stronger structural stability than MET when in complex with L or Q. When K is in a complex with MMP1, the stability of the complex in the human setting is general, but it does not show major unusual fluctuations Fig. .Therefore, L, Q, and K, the active ingredients of SAIN, may play an important role in improving or treating NSCLC (LUAD and LUSC) after acting on the targets EGFR, MET, MMP1, and MMP3.Compared with LUAD, LUSC has a unique pathological morphology  in NSCLC, therefore, no targeted therapy has been developed for LUSC due to few oncogenic aberrations that can target LUSC.Fibroblast growth factors (FGFRs) belong to the family of receptor tyrosine kinases (RTKs) along with EGFR, which comprises the four members FGFR 1–4. In our study, we observed amplification or mutations of FGFR1-4 in NSCLC, which play a crucial role in tumor development and maintenance. In 2016, a gene sequencing study that included 4853 cancer patients revealed that 7.1% of cancer patients had abnormalities in both FGFR 1–4 genes. The most commonly found amplification of the FGFR-1 gene accounted for 50% . A largeMatrix metalloproteinase 3 (MMP3) is the third member of the matrix metalloproteinase (MMP) family, and studies have shown that it plays many roles in healthy humans and patients, such as promoting epithelial mesenchymal transition, increasing the levels or activity of pulmonary profibrotic mediators, or decreasing antifibrotic mediators, and promoting abnormal epithelial cell migration and other abnormal repair processes . In addiIt is well known that insulin enhances the proliferation, migration, and drug resistance of NSCLC cells by activating the PI3K / Akt pathway. In previous studies, it has been shown that insulin can upregulate gene expression of MMP3 in NSCLC cells, and that its expression can be inhibited by PI3K / Akt pathway inhibitors .In previous studies, it has been shown that the genes and proteins of FGFR-1 and MMP3 are highly expressed in esophageal squamous cell carcinoma, and may be associated with esophageal cancer invasion and metastasis .Our findings showed that MMP3 positively regulated NSCLC, especially LUSC by participating in transcriptional misregulation in cancer, the IL-17 signaling pathway, and the rheumatoid arthritis pathway. Furthermore, molecular docking results showed that Q could bind to MMP3 protein with high binding affinity of -8.9 kcal/mol. Therefore, we speculated that there may be consistency or homology between MMP3 and FGFR-1 in treating LUSC. MMP3 may serve as a biomarker for the diagnosis and prognosis of LUSC . HoweverThe latest NSCLC guideline 2021 V2 edition issued by the national comprehensive cancer network (NCCN) indicated that genes associated with targeted therapy of NSCLC include EGFR, KRAS, ALK, ROS1, MET, BRAF, RET, and NTRK . HoweverReceptor tyrosine kinases (RTKs) are high affinity cell surface receptors for many hormones, cytokines, and polypeptide growth factors. Of the 90 unique tyrosine kinase genes identified in the human genome, 58 encode RTK proteins. RTKs have not only been shown to be key regulators of normal cellular processes but also play a key role in the initiation and progression of many types of cancer .EGFR and MET belong to members of the RTK family, and MET, the tyrosine kinase transmembrane receptor for MET, is a protein product encoded by the c-MET proto oncogene that has been of interest in the clinic as a potential therapeutic target in multiple tumor types .Gefitinib, Erlotinib, Afatinib, Osimertinib, Dacomitinib, Lcotinib, and Brigatinib, among others, monoclonal antibodies are comparatively rare and mainly include Necitumumab, Amivantamab, and Ramucirumab, of which Amivantamab targets the EGFR exon20 insertion [Currently, a variety of targeted drugs have been developed for NSCLC targeting EGFR mutations and MET exon14 alterations, among which, drugs targeting EGFR mutations are mainly classified into two major classes: small molecule tyrosine kinase inhibitors (TKI) and monoclonal antibodies, and the TKI class mainly includes nsertion .Crizotinib, Cabozantinib, MGCD265, AMG208, Altiratinib, Golvatinib) and selective MET inhibitors . Monoclonal antibodies can be further divided into Anti-MET antibodies  and anti-HGF antibodies .Drugs targeting alterations in MET exon14 are two major classes of TKIs and monoclonal antibodies, multi kinase inhibitors, including TKIs  had significantly shorter prognostic survival relative to patients with MET mutations (P < 0.02).Smoking confers an increased risk of lung cancer, and in an earlier study, no significant difference by NQO1 genotype was found in a large cohort of patients with combined smoking and nonsmoking lung adenocarcinoma. The data showed that smokers with combined MPO genotype had a lower lung cancer prevalence than nonsmokers .MPO is a member of the heme peroxidase superfamily and is mainly expressed in neutrophils and monocytes . It has High expression of MMP1 has potential in NSCLC treatment.Molecular docking of C-T revealed that components Q, L, and K could bind to MMP1 protein with better affinity than any other component, Q , L , and K , respectively. Our survival analysis of LUAD and LUSC patients after comparing MMP1 overexpression revealed that the overexpression of MMP1 exhibited a worse prognostic survival compared with the survival time of patients after an EGFR mutation. Moreover, PPI network analysis showed that MMP1 formed three triangular structures with EGFR, MMP3, MET, and MPO Fig. d, suggesMMP1, like MMP3, belongs to the MMP family. Only few studies have been performed on MMP1 because the expression of MMP1 in rodent orthologs is not conserved . HoweverTherefore, combined with the above-mentioned results, we speculate that MMP1 may be a novel target for NSCLC treatment, and l, Q, and K may be the relevant lead compounds.In this study, we screened the three active ingredients Q, L, and K from SAIN. However, as to whether these three ingredients have curative effects on NSCLC, experiments in cell systems or animal experiments are needed for further verification. Before performing in vitro and in vivo validation, extensive experimental validation for the treatment of NSCLC on several of these components was found in the literature, therefore, we did not perform in vitro or in vivo validation.L —5,7-dihydroxy-4-chromenone) is a natural flavonoid compound, and it has been reported that the effects of L on NSCLC are mainly reflected in (1) in lung cancer A549 cells and nuclear H460 xenograft mice. The expression of L in melanoma 2 (AIM2) was significantly reduced at the mRNA and protein expression levels, which inhibited AIM2 inflammasome activation, and in turn induced G2/M phase cell cycle arrest and inhibited the epithelial mesenchymal transition (EMT) process in NSCLC . (2) At Q —3,5,7-trihydroxy-4-chromenone) and L are structurally similar, the only difference being that Q has more than one hydroxyl group in position 3 of the chromoenone parent nucleus. Epidemiologically, studies have shown that Q has the effect of preventing lung cancer, which is mainly reflected by (1) Q can significantly enhance tumor necrosis factor-related apoptosis inducing ligand (TRAIL)—induced cytotoxicity in NSCLC cells, thereby accelerating the death of NSCLC cells , 63. (2)In previous studies, it was shown that dietary flavone K —4-chromenone) can effectively prevent and treat lung cancer, mainly through the following mechanisms: (1) K inhibits the apoptosis of H460 cells (NSCLC cells) by inducing the overexpression of tumor suppressor gene antioxidant enzyme (Sod-2) . (2) K mIt is well known that the complexity of NSCLC etiology does not allow us to explain it by the alteration of a single gene, therefore. The treatment of lung cancer is not achieved by one means, but requires combination therapy of multiple means and multiple targets. Although the three components Q, L, and K in SAIN can do so by binding to MMP1, MMP3, EGFR, and MET, and are capable of acting as inhibitors by participating in EGFR tyrosine kinase inhibitor resistance, adherens junction, IL-17 signaling pathway, NSCLC, and the relaxin signaling pathway, exerting their functions together to prevent and treat NSCLC (LUAD and LUSC). Nobody can predict which genetic mutations are responsible for NSCLC, and therefore, a concerted effort from researchers in all fields is required for the complete cure of NSCLC.


In two independent datasets, co-occurrence of NOTCH+/co-DDR+ was also validated to be a better immunotherapy efficacy . By analyzing TCGA cohort, we found patients with coexisting NOTCH+/co-DDR+ pathway had a higher TMB, more infiltration of CD4+T cells. Overall, co-occurrence of NOTCH and co-DDR pathway reflect a better immunotherapy efficacy in advanced NSCLC. This genomic predictor show promise in stratifying patients that suit for immunotherapy for future clinical practice.Although immune checkpoint inhibitors (ICIs) have shown remarkable benefit for treatment of advanced non-small lung cancer (NSCLC), only a minority of patients can achieve durable responses and the most patients produce an ultra-rapid progressive disease. Here, we collected the availably published datasets and mined the determinants of response to immunotherapy on pathway levels. One hundred six NSCLC patients treated with immunotherapy were combined from Rizvi et al. and Hellman et al. studies (whole exon sequencing). Two independent validation datasets consisted of the MSKCC cohort (targeted sequencing) and data by Anagnostou and colleagues (whole exon sequencing). The Cancer Genome Atlas (TCGA) somatic mutation and gene expression data were applied to explore the immunobiology features. In the first combined cohort, we detected NOTCH pathway altered in 71% patients with durable clinical benefit (DCB) while only 36% among no durable benefit (NDB) (p = 0.005). Compared to NDB group, co-occurrence of NOTCH and at least two DDR (co-DDR) pathway was discovered in DCB group and contributed to a prolonged progression-free survival (PFS) [22.1  Immune checkpoint inhibitors (ICIs) has shown remarkable benefit for treatment of advanced non-small lung cancer (NSCLC) , 2. NeveA plethora of studies have shown biomarkers for predicting response to immunotherapy. The most heavily studied biomarker was programmed death-ligand 1 (PD-L1) expression correlated to efficacious immunotherapy –8, but sDue to the interactions and dependencies among different pathways, we supposed single pathway was not high enough to reflect the response of ICI. Therefore, we focused on the co-occurrent pathways to predict the superior survival outcomes after immunotherapy. Availably published datasets were collected from four studies. The first cohort was combined by data from Rizvi et al. and Hellman et al., which was used to identify the co-occurrent pathways correlated to ICI , 19. We http://www.cbioportal.org/study/summary?id=tmb_mskcc_2018). We also acquired the genome mutation from 1026 NSCLC patients and transcriptome expression data from 59 advanced NSCLC patients in TCGA.We collected four publicly available datasets treated with immunotherapy  of immune cell types in the tumor microenvironment . Gene siWe performed univariable and multivariable Cox regression analyses to identify potential predictors of survival. Survival curves were estimated with the Kaplan-Meier product-limit method and compared by log-rank test. Comparisons of TMB, TNB, and expression levels between different groups were used with the Wilcoxon rank-sum test or t test. Enrichment analysis of gene function was calculated by GSEA with Benjamini-Hochberg correction for multiple hypothesis testing (q < 0.05). All statistical analyses were performed in the R statistical environment version 3.6.2.vs 82%), followed by HIPPO pathway (65% in DCB vs 52% in NDB). Notably, 71% patients with DCB had alteration in NOTCH pathway while only 36% in NDB group (p = 0.005). This was consistent with previous study that NOTCH signaling correlated with better ICI efficacy . Taken together, alterations on pathway level can reflect the divergent response in immunotherapy.We collected WES data from 106 NSCLC patients treated with immunotherapy derived from published data as the discovery cohort , 19. Of efficacy . More DDefficacy , we alsovs 3.6 months, p < 0.0001, versus SinglePath  versus wild type  (versus NOTCH or co-DDR occurrence versus Wildtype), co-occurrence of NOTCH/co-DDR also exhibited superior survival outcomes and the highest TMB  and 45 Lung squamous cell carcinoma (LUSC)] for further analyses . Co-occuversus BadBenefit group or among three groups . Of note, CD4+ T cell were enriched in NOTCH+/co-DDR+ group  method was employed to deconvolute the relative abundance of each immune cell type. By unsupervised clustering, we classified the 59 NSCLC into three clusters [immune-high (n = 27), immune-intermediate (n = 15), and immune-low (n = 17), R+ group . AnotherR+ group . FunctioIn this study, we demonstrated the co-occurrence of NOTCH and co-DDR pathways can predict the superior survival outcomes after immunotherapy in advanced NSCLC. This phenomenon was first identified in one combined cohort and validated across another two independent cohorts. All the data we analyzed was performed WES, except one validated cohort from MSKCC (targeted sequencing). Using WES data was an advantage because it can cover enough genes of the pathway, overcoming the deficiency of targeted sequencing. To the best of our knowledge, this is the first report focusing on co-occurrence event on pathway level to explore the determinants of response to ICIs.To explore whether there were dominant gene mutations in the co-occurring pathway, we depicted the co-occurring event on gene level involved in NOTCH and DDR pathways  or wild type, and the TMB was gradually decreasing. As for the validation dataset of cohort 2, we also observed the longer PFS in CoPath group than SinglePath, but there was no significant different in SinglePath Patients carrying alterations of NOTCH+/co-DDR+ pathway associated with increased TMB, TNB, and more infiltration of CD4+ T cell. Previous study has revealed that alterations in co-DDR pathway appeared to higher TMB and TNB . Since aIn conclusion, preliminary data from three cohorts showed evidence that co-mutations in NOTCH and co-DDR pathways indicated better immunotherapy efficacy. This genomic marker provided a new dimension to predict the superior survival outcomes in response to immunotherapy.The original contributions presented in the study are included in the article/CC, CS, and ZZ conceived and designed the present study. ZZ, YYG, XS, and JB contributed equally to this work. ZZ and CC wrote the manuscript. ZZ, YYG, and XS collected and interpreted data. JB and WG performed the data analysis. JM, JL, and XX gave material support. JH, BZ, and YFG helped in improving the language and correcting grammar mistakes. All authors contributed to the article and approved the submitted version.China Postdoctoral Science Foundation funded project (Grant #2014T70977). Hubei Province National Natural Science Foundation of China (Grant #2018CFB733).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


It can be argued that it has never been more exciting than right now to be researching Molecular Signalling and Pathways in Neuroscience. This is the study devoted to identifying molecular and cellular players that underlie the structure, design and function of the brain across all levels. This study is not new and evolved with the major discoveries of biological sciences, including those discoveries that led to our fundamental understanding that genetic information is encoded by DNA, that is transcribed to RNA, that is translated to proteins, that in turn organise themselves together with other cellular biomolecules to carry out fundamental tasks of the cell. It is important to point out that signalling is a concept that existed prior to the discovery of DNA's genetic code. Indeed, work from the study of hormones had already pointed to the notion that processes within the cell could be influenced by the binding of ligands to cell surface receptors and this could then lead to important changes in tissues and organisms. Perhaps one of the most emblematic examples of such discoveries in neuroscience is the report in 1951 of a factor in tumour extracts that can cause neurite outgrowth and that was therefore aptly called nerve growth factor  . Screening results, or Omics profiles, can be used for Dissecting out the intricacies of signalling pathways at the molecular level is fascinating in its own right; however, it becomes downright exciting when pathways are linked to the functioning of the nervous system in health and disease, with a particular focus on synaptic and cellular proteins that define cellular identities. This is of course the element that gives the Molecular Signalling and Pathways in Neuroscience challenge all of its purpose. How do individual steps in signalling pathways lead to functional effects in cells and organisms? In many cases, clues will have arisen from molecular studies as to which phenotypic effects may be expected from a specific signalling cascade. Alternatively, phenotypic screening will be required to determine suitable phenotypes to test in relation to a specific signalling pathway. In either case, a multitude of molecular and pharmacological approaches are at our disposal to explore the pathway-phenotype relationships. Individual proteins can be modified in model systems , including expression of mutant forms of the proteins (exogenous expression or knock-in models) or gene knock-outs or knock-downs, and the effect of this modification measured on specific phenotypes, for example how the knock-out of a synaptic protein would affect synaptic vesicle release or neurite complexity in primary neurons. In addition, pharmacological approaches exist, for instance when tool compounds, existing or newly developed, are used to activate or inactivate a protein within the signalling cascade, when it is again possible to explore how this modification modifies the pathway's phenotypes. These few examples given here are again not limitative relative to our challenge. The activity of linking pathways to phenotypes is in continuous progression both at the scientific and at the technological level and this should be reflected in the submissions to this section. The pathway-phenotype relationships are much more than the icing on the cake of Molecular Signalling and Pathways in Neuroscience. It is precisely this relationship that allows us to go further both for fundamental studies and for applied research. Indeed, it allows further dissection of pathways, such as uncharacterized partners within these pathways, specific post-translational modifications within pathways or the phenotypes of pathways in specific conditions. It also allows development of tools to modulate pathway phenotypes such as developing pharmacological agents targeting specific elements of pathways such as key enzymes, receptors or protein-protein interactions. Linked to this, the understanding of pathway-phenotype relationships allow the possibility to develop models for this pathway that can again have fundamental scientific goals or destined for specific applications such as disease modelling.As the old saying goes, we have no idea what the future has in store for us. However, of one thing we can be certain: we have never been in a better position to make leaping advances in Molecular Neuroscience as today. With amazing resources available to us such as whole genomes and proteomes of multiple species, nervous system expression patterns of just about any gene, a broad and expanding range of techniques for molecular manipulation of cells and model organisms, idem for pharmacological tools, a broad and expanding range of technologies to characterise cellular and organismal phenotypes, it is both our duty and our privilege to address the challenge of elucidating Molecular Signalling and Pathways in Neuroscience. No doubt that scientists will be hard at work to make this happen and we can look forward in hopeful anticipation to many exciting studies to come in this field.J-MT: wrote the manuscript.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Scutellariae radix-Coptidis rhizoma drug pair (SCDP) in ulcerative colitis (UC) by network pharmacology and molecular docking and to explore the associated therapeutic mechanism. The effective active ingredients and targets of SCDP were determined from the TCMSP database, and the drug ingredient-target network was constructed using the Cytoscape software. The disease targets related to UC were searched in GeneCards, DisGeNET, OMIM, and DrugBank databases. Then, the drug ingredient and disease targets were intersected to construct a protein-protein interaction network through the STRING database. The Metascape database was used for the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of the predicted targets of SCDP for UC. The Autodock software was used for molecular docking between the main active ingredient and the core target to evaluate the binding ability. SCDP has 43 effective active ingredients and 134 intersection targets. Core targets included AKT1, TP53, IL-6, VEGFA, CASP3, JUN, TNF, MYC, EGFR, and PTGS2. GO functional enrichment analysis showed that biological process was mainly associated with a cytokine-mediated signaling pathway, response to an inorganic substance, response to a toxic substance, response to lipopolysaccharide, reactive oxygen species metabolic process, positive regulation of cell death, apoptotic signaling pathway, and response to wounding. KEGG enrichment analysis showed main pathway concentrations were related to pathways in cancer, AGE-RAGE signaling pathway in diabetic complications, bladder cancer, IL-17 signaling pathway, apoptosis, p53 signaling pathway, and PI3K-Akt signaling pathway. The drug active ingredient-core target-key pathway network contains 41 nodes and 108 edges, of which quercetin, wogonin, baicalein, acacetin, oroxylin A, and beta-sitosterol are important active ingredients; PTGS2, CASP3, TP53, IL-6, TNF, and AKT1 are important targets; and the pathways involved in UC treatment include pathways in cancer, PI3K-Akt signaling pathway, AGE-RAGE signaling pathway in diabetic, apoptosis, IL-17 signaling pathway and herpes simplex infection. The active ingredient has a good binding capacity to the core target. SCDP key active ingredients are mainly quercetin, wogonin, baicalein, acacetin, oroxylin A, and beta-sitosterol, which function mainly by regulating targets, such as PTGS2, CASP3, TP53, IL-6, TNF, and AKT1, and are associated with multiple signaling pathways as pathways in cancer, PI3K-Akt signaling pathway, apoptosis, IL-17 signaling pathways.This study aims to analyze the targets of the effective active ingredients of  It is an incurable disease with low mortality . The incnt of UC , 9. Undent of UC , 11.Scutellariae radix-Coptidis rhizoma drug pair is the main ingredient herb for clearing away heat and dampness. Their compatibility is often used to treat damp-heat syndromes, dysentery, diarrhea, and other diseases [Scutellaria baicalensis and Coptis chinensis is mainly responsible for clearing away heat and dampness and has an antidiarrheal and antidysentery efficacy. Furthermore, Hu et al. used a database to analyze the medication regularity of TCM compounds in the treatment of UC. Results showed that Scutellaria baicalensis and Coptis chinensis were among the top 26 drugs with the highest medication, of which and Coptis chinensis was the most frequently used drug [Scutellaria baicalensis, Coptis chinensis, Phellodendron amurense, and Sophora flavescens can be used together and they are particularly suitable for patients with the damp-heat syndrome, such as abdominal bloat, bitter mouth, nausea, vomiting, and/or yellow tongue coating [Scutellaria baicalensis and Coptis chinensis are the drugs which were used more frequently in the treatment of UC and are commonly paired drugs to treat damp-heat intrinsic syndrome [diseases –14. Furtdiseases –18. The sed drug . Profess coating . Previousyndrome , 22.Scutellariae radix-Coptidis rhizoma drug pair (SCDP) in the treatment of UC is that baicalein can reduce the number of apoptotic intestinal epithelial cells due to its ability to reduce the expression of the 78-kDa glucose-regulated protein (GRP78) and caspase-3 [κB), reduce the expression of the proinflammatory cytokines IL-1β, IL-6, matrix metalloproteinase-9, chemokine motif receptor 1 (CX3CR1), cyclooxygenase-2 (COX-2), and telomerase reverse transcriptase, and increase the expression of the tight junction scaffolding protein zonula occludens-1, to play a therapeutic role in UC [Scutellariae radix and Coptidis rhizoma, exert therapeutic effects by inhibiting the expression of toll-like receptor 4 (TLR4)/NF-κB pathway and suppressing the inflammatory factors secretion as TNF-α, IL-6, IL-1β, and IL-4 in the colon of UC animal models [Coptidis rhizoma, can exert a therapeutic effect on UC through increasing glutathione peroxidase and superoxide dismutase contents and inhibiting oxidative stress response as well as apoptosis [One proposed mechanism of the aspase-3 . Researcle in UC . Gegen Ql models . In addipoptosis .Scutellariae radix-Coptidis rhizoma drug pair in the treatment of UC was analyzed by network pharmacology technology and molecular docking approaches to help better guide clinical treatment and mechanism research.Network pharmacology and molecular docking are new technologies based on systems biology and databased molecular correlation analysis in the exploration of new drugs and prediction of drug targets. Network pharmacology emphasizes multi-ingredient-multitargets-multipathway regulation of the signal pathway. Many studies have confirmed that network pharmacology is significant for determining the target of drugs and screening the active ingredients of drugs –27. In thttps://tcmspw.com/tcmsp.php) was used for the active ingredient screening. Oral bioavailability (OB) ≥30% and drug-likeness (DL) ≥0.18 were used as absorption, distribution, metabolism, and excretion parameters. Target genes were predicted using TCMSP and the Swiss Target Prediction database (http://www.swisstargetprediction.ch/) after identifying the active ingredients. The names of the predicted target genes were standardized from the UniProt database (https://www.uniprot.org/). The SCDP ingredients and target gene network were drawn using the Cytoscape software, and the top 10 active ingredients were identified by their degree values.The Traditional Chinese Medicine Systems Pharmacology (TCMSP) database and analysis platform (https://www.genecards.org/), DisGeNET (https://www.disgenet.org/), OMIM (http://www.omim.org/), and DrugBank (https://www.drugbank.ca/) were selected to obtain UC-related pathogenic genes. Both databases can be used to find the most cutting-edge disease-related genes. All the databases were used “ulcerative colitis” as the keyword.GeneCards (http://string-db.org/) with the species limited to “Homo sapiens” and a confidence score of >0.4. Next, the interaction files were downloaded and imported into Cytoscape to visualize the PPI network and analyze the topological value. Then, based on the MCODE plug-in, the targets with a median value and a median value that was not less than the average value were recognized as the key targets of SCDP for the treatment of UC. And the top 10 ranked proteins were defined as hub targets based on the degree level.PPI is particularly important for biological process analyses and is vital for understanding complex mechanisms in a living cell. PPI network mapping was performed on the obtained bioactive ingredients and disease targets using the Search Tool for the Retrieval of Interacting Genes (STRING) database (http://metascape.org/) for GO and KEGG enrichment analyses by entering the list of intersection target gene names and selecting the species as “Homo sapiens” for customized analysis with a cutoff P value of <0.01. According to the results, a network visualization was constructed by the tools in the bioinformatics website (http://www.bioinformatics.com.cn).The predicted targets were imported into the Metascape database  and PDB (https://www.rcsb.org) databases.Molecular docking is used to interpret the binding area of small molecule ligands and macromolecular receptors through computer simulation and then calculates the physical and chemical parameters for predicting the affinity between the two. Receptor and ligand files were downloaded from PubChem  protein complex, ; (2) functional module . The MCODE plug-in was then applied to analyze the interactions using a molecular complex detection algorithm, and 3 module clusters were obtained . AccordiMetascape was used for GO and KEGG analyses of the 134 hub genes to obtain enriched ontology clusters. As a result, we obtained 2304 GO terms, including 2073 biological processes (BPs), 79 cellular components (CCs), and 152 molecular functions (MFs). KEGG pathway enrichment analysis screening resulted in 164 signaling pathways. Visual analysis of the top 20 pathways showed that the main signaling pathways of SCDP in the treatment of UC were concentrated in pathways in cancer, the advanced glycation end products and the receptor for AGEs (AGE-RAGE) signaling pathway in diabetic complications, bladder cancer, IL-17 signaling pathway, apoptosis, p53 signaling pathway, and phosphoinositide 3-kinase (PI3K)-Akt signaling pathway .The Cytoscape software was used to construct an active ingredient-core target-key pathway network, which contained 41 nodes and 108 edges . AccordiThe molecular docking results indicate binding energies of less than −2 kcal/mol after docking, suggesting the formation of stable bonds between the active ingredients and predicted target proteins . The PyMUC is a type of irritable bowel syndrome (IBS), and the main pathogenesis of UC is spleen deficiency as the original cause and damp-heat with stasis toxin as the superficial factors in TCM theory –30. BaseSCDP is a commonly used drug pair for eliminating damp-heat toxin. These two herbs are also constituents of many classical formulas, such as decoctions Gegen Qinlian, Huangqin, Shaoyao, Baitouweng, Huanglian Jiedu, Banxia Xiexin, and WuMei pill, for the treatment of diarrhea as a symptom of UC, by TCMs .Scutellariae radix, Coptidis rhizoma, Cortex Phellodendri, Sophora flavescens, pulsatilla, ash bark, among others, in combination with dampness-dissipating drugs, such as Pogostemon cablin and eupatorium, while Y. Zhang et al. suggested the use of Coptidis rhizoma, Scutellariae radix, Cortex Phellodendri, Sophora flavescens, and pulsatilla as the main ingredients in the formulation [The damp-heat insidious pathogen has been proposed as the mechanism responsible for UC recurrence, necessitating that damp-heat toxin is cleared for UC treatment. In turn, Cao et al. advocated the use of mulation , 33, 34.α and lipocalin-2 mRNA and enhancing the expression of the Slip protein to protect the UC mucosa [α, IL-6, and increasing the expression of IL-10 [The results show that the key active ingredients are quercetin, wogonin, baicalein, acacetin, oroxylin A, and beta-sitosterol. Previous research discovered that quercetin inhibits the expression of LPS-induced inflammatory genes, mainly by reducing the levels of TNF-C mucosa . It has of IL-10 .α to regulate inflammatory responses [κB pathway in Caco-2 cells, suggesting its protective effect on the intestinal mucosal barrier [Wogonin has strong anti-inflammatory, antitumor, antiviral and antiallergic properties. It can inhibit the activity of Treg cells, regulate the differentiation of Th17 cells, and inhibit the transcription of the extracellular signal-regulated kinase (ERK), signal transducer and activator of transcription 3 (STAT3), and hypoxia-inducible factor-1esponses . One stu barrier .Coptidis rhizoma. Studies have suggested its anti-inflammatory effect and discovered that it could improve the inflammatory state of UC and the control drugs [κB p65 and increases the level of IκBα to resists the pathological changes of dextran sulfate sodium (DSS)-induced UC [Baicalein is the main ingredient of ol drugs –41. The ol drugs , 43. In duced UC .Acacetin  is an O-methylated flavone naturally present in plants like chrysanthemum and safflower, as well as in Calamintha and Linaria species. Acacetin possesses antiperoxidative, anti-inflammatory, antiplasmodial, and anticancer activities. In addition, acacetin strongly inhibits the expression of proinflammatory cytokines, inducible nitric oxide synthase (iNOS), and COX-2 in LPS-induced RAW 264.7 cells . ResearcκB downstream target genes and inhibits the transcription of downstream target genes of Nrf2 [Oroxylin A and baicalein are produced as major metabolites of baicalin by the action of bacteria in the large intestine cavity. In addition, both baicalin and baicalein are converted into four new metabolites: wogonin, wogonoside, oroxylin A, and oroxin A . Some st of Nrf2 –49.α, IL-6, and IL-1β levels in the intestinal tissues of experimental colitis mice in a concentration-dependent manner [Beta-sitosterol was found to markedly reduce weight loss, colonic length shortening, and microscopic changes in DSS-induced colitis and to reduce TNF-t manner .κB and oxidative stress pathways. An important target of anti-inflammatory drugs, especially nonsteroidal anti-inflammatory drugs, is prostaglandin endoperoxide synthase or COX. There are two subtypes of COX. COX1 is constitutively expressed and is responsible for regulating normal physiological processes. COX2 is induced under inflammatory conditions and mainly regulates the inflammatory process. It has been suggested that the expression of COX2 is induced in the colon of IBD patients but also in the inflamed tissues of IL-10-deficient IBD mouse models, and reduced COX2 is considered a major target for IBD treatment [The core targets of SCDP in UC treatment are PTGS2 (COX2), CASP3, TP53, IL-6, TNF, and AKT1, which are associated with the TNF signaling pathway and NF-reatment .A previous study in UC rats revealed that baicalin could up-regulate B-cell lymphoma 2 (Bcl-2) protein expression, lower Bax  protein expression, induce marked activation of the caspase-cascade system, and exhibit concentration-dependent down-regulation of Fas and Fas Ligand (FasL) protein expressions to exert therapeutic effects in UC . AKT1 isκB signaling pathway [Berberine has been found to play a regulatory role in macrophage M1 polarization in DSS-induced colitis through the AKT1-suppressor of cytokine signaling-1 (SOCS1)-NF- pathway . IL-6 ge pathway . JUN is  pathway . Being a pathway , 57.GO analysis and KEGG enrichment results showed that the mechanism of SCDP in the treatment of UC was mainly related to the following pathways: pathways in cancer, PI3K-Akt signaling pathway, the AGE-RAGE signaling pathway in diabetic complications, apoptosis, IL-17 signaling pathway, and herpes simplex infection.According to Rogler, up to 30% of the patients with chronic active UC or Crohn's disease (CD) are likely to get colorectal cancer after 35 years since the onset of the disease . Chronicα, and IL-1B), chemokines (CXC), granulocyte colony-stimulating factor, and granulocyte-macrophage colony-stimulating factor. Meanwhile, IL-17A is also involved in the regulation of mucosal barrier function and amplification of the complete inflammatory response through the production of antimicrobial peptides, such as β-defensins, S100 β-defensins, and S100 proteins [IL-17, mainly secreted by Th17 cells, is involved in inflammatory response, and its levels have been confirmed to correlate with the extent of UC disease. Acting as a proinflammatory factor, IL-23 promotes IL-17 production and forms the IL-23/17 axis to amplify the inflammatory response , 61. IL-proteins . The strproteins , 64. OthThe mutation of the p53 gene is an important genetic alteration involved in the early stages of UC-associated colorectal cancer. The overexpression of p53 protein in the colorectal crypt of UC patients, usually in the absence of atypical hyperplasia observation of atypical hyperplasia, is used by pathologists to define the state between regenerative changes and intraepithelial neoplasia. It has been used as a biomarker in predicting the risk of progression into a malignant tumor. High-frequency mutations of p53 were reported in severe chronic UC patients with undiagnosed cancer . The expα and IL-1 and the upregulation of IL-6 and IL-10. The PI3K-AKT signaling pathway is involved in the regulation and release of proinflammatory cytokines, including TNF-α and other cytokines closely related to the inflammatory response of UC [β (TGF-β) expression and lead to a decrease in PI3K/AKT activation, thereby alleviating the symptoms of UC by regulating TGF-β expression [κB, leading to inflammatory responses, cell apoptosis, and microvascular diseases [κB and its response to oxidative stress [In addition, TCMs may exert relevant apoptotic inhibitory effects through the upregulation of p53 and Bcl-2 expression, which is accompanied by the downregulation of TNF-se of UC , and plapression . AGE anddiseases . RAGE poe stress .PTGS2, CASP3, TP53, IL-6, TNF, and AKT1, suggesting that the key active ingredients of SCDP play a therapeutic role in UC by mainly intervening with the key signaling targets, such as the inflammatory response and oxidative stress. In summary, the potential biological mechanism of SCDP in the treatment of UC involves multiple ingredients, targets, and pathways, and the therapeutic effects occur through several pathways as pathways in cancer, PI3K-Akt signaling pathway, apoptosis, and IL-17 signaling pathway.The results of molecular docking showed that quercetin, wogonin, baicalein, acacetin, oroxylin A, and beta-sitosterol, the main active ingredients of SCDP, had a good affinity to the core target genes The presented study provides a scientific basis for the subsequent development and utilization of SCDP. Even so, network pharmacology has only analyzed the main active ingredients and targets of drugs, and further validation of their predicted target pathways is needed experimentally. The conclusions of this research also contribute to the research and development of drugs and further elaboration of mechanisms.