menopause is associated with increased risk of metabolic diseases such as type 2 diabetes and metabolic syndrome ( 1 , 2 ) . dyslipidemia , obesity , and insulin resistance are of the most common features of the metabolic syndrome in menopausal woman and known to be important risk factors for cardiovascular diseases ( 3 - 5 ) . menopause is also associated with a wide range of changes in the expression and secretion of proinflammatory cytokines which may explain increased incidence of osteoporosis , alzheimer , arthritis , and other inflammatory diseases seen in a postmenopausal period ( 6 - 9 ) . for many years , hormone therapy was the first primary treatment modality for the management of menopause ; however , it had own limitations like increasing risk of stroke , coronary heart disease , pulmonary embolism , and ovarian cancer ( 1 , 10 ) . therefore , development of new agents with anti - inflammatory action and lesser side effects for management of menopause remains an attractive subject . it has been reported that a number of medicinal plants may be useful for preventing or treating some menopausal related complications such as insulin resistance , dyslipidemia , hepatic steatosis , weight gain , inflammation , and loss of bone mass ( 11 - 13 ) . trigonella foenum - graecum ( fenugreek ) , a plant of fabaceae family , has a long history for management of diabetes and dyslipidemia ( 14 , 15 ) . several studies also support anti - inflammatory and immunomodulatory activities of t. foenum - graecum in adult male animals ( 16 - 18 ) . recently , sindhu et al . ( 19 ) reported that mucilage extracted from ethanolic extract ( ee ) of t. foenum graecum has anti - inflammatory and antioxidative properties on adjuvant - induced arthritis in female rats . further , hakimi et al . showed that t. foenum graecum seed decreases the number of hot flashes and vasomotor symptoms in menopausal women ( 20 ) . however , to the best of our knowledge , no study has yet evaluated the effect of this plant on the level of proinflammatory cytokines in menopause . the present work was performed to investigate the effects of chronic treatment with t. foenum graecum seed extracts on serum concentration of interleukin-1 ( il-1 ) , interleukin-6 ( il-6 ) and tumor necrosis factor- ( tnf- ) in ovariectomized rats . the seeds of t. foenum - graecum were purchased from local market ( guilan , iran ) and identified at the herbarium of agricultural research center of guilan university ( iran ) . the ee and hexanic extract ( he ) of t. foenum - graecum were prepared using a maceration method as described previously ( 21 , 22 ) . briefly , the air - dried powder of the seeds was suspended in 96% ethanol ( 5 ml per g of seed ) and maintained for 72 hour at 37c . then the solvent was removed and a new extraction started again with fresh ethanol . finally , the ee was filtered through whatman no.1 filter paper and dried using a rotary vacuum evaporator ( glf , germany ) at 50c . female wistar rats initially weighing 190 to 220 g were obtained from pasteur institute , hesarak branch , iran . the animals were maintained at 22c  4c on a light / dark cycle of 12 hour and away from stress . at all stages of the study , they had free access to enough laboratory chow and tap water ad libitum . ethical rules of working with laboratory animals were approved by guilan university of medical sciences . after two weeks acclimatization period , animals were randomly divided into the following 7 experimental groups , each group including 7 animals : group i included sham control rats ; group ii comprised ovariectomized control animals ; groups iii and iv included ovariectomized animals and administrated intraperitoneally ee of t. foenum - graecum ( 50 and 150 mg / kg , respectively ) ; groups v and vi comprised ovariectomized animals and treated intraperitoneally he of t. foenum - graecum ( 50 and 150 mg / kg , respectively ) ; groups vii included ovariectomized positive control rats , which administrated orally 10 g / kg of 17- estradiol valerate ( aburaihan pharmaceutical co. , iran ) . the treatments were started one day after removal of ovaries and lasted for 42 days . during this period , fasting blood glucose ( fbg ) and body weight of all of the rats were recorded prior to the start and at the end of treatments . after an overnight fast , each rat was anesthetized with intraperitoneally injection of 320 mg / kg chloralhydrate ( arya pharmaceutical co. , iran ) ( 23 ) . then , a small abdominal skin incision was performed and after exposing ovaries , they were removed bilaterally and tubal ends were closed using 2/0 silk string to prevent internal bleeding . then the fat pad was repositioned into the abdomen and muscle and skin the animals in the control group underwent a similar procedure without eliminating the ovaries . at the end of the 42th day , the samples were centrifuged for 5 minutes at 3000 rpm and supernatants were maintained at -20c for further analysis . the level of fbg was determined using accu - check glucometer ( roche diagnostic , germany ) . concentrations of il-1 , il-6 and tnf- were measured by e0071ra , e90079ra , and e90133ra elisa kit , respectively ( life science inc , china ) . differences between means of different groups were analyzed using one - way analysis of variance ( anova ) followed by tukey s post hoc test for multiple comparisons . paired t - test was used to compare the differences between day 0 and day 42 . results were exhibited as the mean  standard error of the mean ( sem ) . the seeds of t. foenum - graecum were purchased from local market ( guilan , iran ) and identified at the herbarium of agricultural research center of guilan university ( iran ) . the ee and hexanic extract ( he ) of t. foenum - graecum were prepared using a maceration method as described previously ( 21 , 22 ) . briefly , the air - dried powder of the seeds was suspended in 96% ethanol ( 5 ml per g of seed ) and maintained for 72 hour at 37c . then the solvent was removed and a new extraction started again with fresh ethanol . finally , the ee was filtered through whatman no.1 filter paper and dried using a rotary vacuum evaporator ( glf , germany ) at 50c . female wistar rats initially weighing 190 to 220 g were obtained from pasteur institute , hesarak branch , iran . the animals were maintained at 22c  4c on a light / dark cycle of 12 hour and away from stress . at all stages of the study , they had free access to enough laboratory chow and tap water ad libitum . ethical rules of working with laboratory animals were approved by guilan university of medical sciences . after two weeks acclimatization period , animals were randomly divided into the following 7 experimental groups , each group including 7 animals : group i included sham control rats ; group ii comprised ovariectomized control animals ; groups iii and iv included ovariectomized animals and administrated intraperitoneally ee of t. foenum - graecum ( 50 and 150 mg / kg , respectively ) ; groups v and vi comprised ovariectomized animals and treated intraperitoneally he of t. foenum - graecum ( 50 and 150 mg / kg , respectively ) ; groups vii included ovariectomized positive control rats , which administrated orally 10 g / kg of 17- estradiol valerate ( aburaihan pharmaceutical co. , iran ) . the treatments were started one day after removal of ovaries and lasted for 42 days . during this period , fasting blood glucose ( fbg ) and body weight of all of the rats were recorded prior to the start and at the end of treatments . after an overnight fast , each rat was anesthetized with intraperitoneally injection of 320 mg / kg chloralhydrate ( arya pharmaceutical co. , iran ) ( 23 ) . then , a small abdominal skin incision was performed and after exposing ovaries , they were removed bilaterally and tubal ends were closed using 2/0 silk string to prevent internal bleeding . then the fat pad was repositioned into the abdomen and muscle and skin the animal was placed in a warm place to come intellect . the animals in the control group underwent a similar procedure without eliminating the ovaries . at the end of the 42th day , blood sample was collected from tail vein of fasted animals for biochemical assays . the samples were centrifuged for 5 minutes at 3000 rpm and supernatants were maintained at -20c for further analysis . the level of fbg was determined using accu - check glucometer ( roche diagnostic , germany ) . concentrations of il-1 , il-6 and tnf- were measured by e0071ra , e90079ra , and e90133ra elisa kit , respectively ( life science inc , china ) . differences between means of different groups were analyzed using one - way analysis of variance ( anova ) followed by tukey s post hoc test for multiple comparisons . paired t - test was used to compare the differences between day 0 and day 42 . results were exhibited as the mean  standard error of the mean ( sem ) . in nonovariectomized sham group , the mean fbg on day 0 ( 5.72  0.12 mmol / l ) and day 42 ( 5.38  0.11 on the other hand , concentration of fbg at the end of the study was significantly higher than day 0 in ovariectomized control animals ( 6.16  0.13 mmol / l vs. 8.77  0.47 mmol / l , p < 0.05 ) . although the administration of estradiol or t. foenum - graecum ( 50 and 150 mg / dl of he and 150 mg / kg of ee ) to ovariectomized animals significantly diminished ( p < 0.05 ) the increase of fbg , in all treated groups the level of fbg was still higher than that of day 0 ( table 1 ) . abbreviations : ee , ethanolic extract ; he , hexanic extract ; and ova , ovariectomy . there was a significant difference regarding weight gain between the sham control ( 53  9 g ) and ovariectomized control groups ( 111  7 g , p < 0.05 ) . estradiol and both extracts of t. foenum - graecum significantly decreased the body - weight gain compared to the ovariectomized control group ( p < 0.05 ) . as shown in figure 1 , the serum il-1 level of the saline - injected rats in the ovariectomized group was significantly higher than those of the sham animals ( 990  2.9 vs. 300  2.8 pg / ml , p < 0.05 ) . administration of both he and ee of t. foenum - graecum significantly decreased serum il-1 in the ovariectomized rats compared to the saline group ( p < 0.05 ) . the level of il-1 in groups received 50 and 150 mg / kg of he was 440  2.8 pg / ml and 410  2.2 pg / ml , respectively . also , concentrations of il-1 in animals treated with 50 and 150 mg / kg of ee were 410  2.2 pg / ml and 610  2.7 pg / ml , respectively . the effect of he was comparable to that of induced by 10 g / kg of estradiol ( 389  2.9 pg / ml ) . the animals were subjected to ovariectomy surgery and then treated intraperitoneally with 10 g / kg estradiol or concentrations of 50 mg / kg and 150 mg / kg of he and ee of t. foenum - graecum seeds for 6 weeks . * p < 0.05 vs. control group ; # p < 0.05 vs. saline group . similarly , serum il-6 significantly increased in ovariectomized control group when compared to the sham animals ( 1258  2.7 vs. 483  3 pg / ml , p < 0.05 ) . as shown in figure 2 , hexanic and ethanolic extracts of t. foenum - graecum , like estradiol , diminished the il-6 level in ovariectomized animals ( p < 0.05 as compared with saline group ) . treatment with 50 and 150 mg / kg of he was also able to significantly decrease serum tnf- from 999  2.4 pg / ml ( saline group ) to 457  2.5 and 445  2.6 pg / ml , respectively , comparable to the effect of estradiol ( 430  2.6 pg / ml ) . a similar decrease in tnf- concentration was induced by both doses of ee ( figure 3 ) . the animals were subjected to ovariectomy surgery and then treated intraperitoneally with 10 g / kg estradiol or concentrations of 50 mg / kg and 150 mg / kg ofhe and ee of t. foenum - graecum seeds for 6 weeks . * p < 0.05 vs. control group ; # p < 0.05 vs. saline group . the animals were subjected to ovariectomy surgery and then treated intraperitoneally with 10 g / kg estradiol or concentrations of 50 mg / kg and 150 mg / kg of he and ee of t. foenum - graecum seeds for 6 weeks . * p < 0.05 vs. control group ; # p < 0.05 vs. saline group . in nonovariectomized sham group , the mean fbg on day 0 ( 5.72  0.12 mmol / l ) and day 42 ( 5.38  0.11 on the other hand , concentration of fbg at the end of the study was significantly higher than day 0 in ovariectomized control animals ( 6.16  0.13 mmol / l vs. 8.77  0.47 mmol / l , p < 0.05 ) . although the administration of estradiol or t. foenum - graecum ( 50 and 150 mg / dl of he and 150 mg / kg of ee ) to ovariectomized animals significantly diminished ( p < 0.05 ) the increase of fbg , in all treated groups the level of fbg was still higher than that of day 0 ( table 1 ) . abbreviations : ee , ethanolic extract ; he , hexanic extract ; and ova , ovariectomy . there was a significant difference regarding weight gain between the sham control ( 53  9 g ) and ovariectomized control groups ( 111  7 g , p < 0.05 ) . estradiol and both extracts of t. foenum - graecum significantly decreased the body - weight gain compared to the ovariectomized control group ( p < 0.05 ) . as shown in figure 1 , the serum il-1 level of the saline - injected rats in the ovariectomized group was significantly higher than those of the sham animals ( 990  2.9 vs. 300  2.8 pg / ml , p < 0.05 ) . administration of both he and ee of t. foenum - graecum significantly decreased serum il-1 in the ovariectomized rats compared to the saline group ( p < 0.05 ) . the level of il-1 in groups received 50 and 150 mg / kg of he was 440  2.8 pg / ml and 410  2.2 pg / ml , respectively . also , concentrations of il-1 in animals treated with 50 and 150 mg / kg of ee were 410  2.2 pg / ml and 610  2.7 pg / ml , respectively . the effect of he was comparable to that of induced by 10 g / kg of estradiol ( 389  2.9 pg / ml ) . the animals were subjected to ovariectomy surgery and then treated intraperitoneally with 10 g / kg estradiol or concentrations of 50 mg / kg and 150 mg / kg of he and ee of t. foenum - graecum seeds for 6 weeks . * p < 0.05 vs. control group ; # p < 0.05 vs. saline group . similarly , serum il-6 significantly increased in ovariectomized control group when compared to the sham animals ( 1258  2.7 vs. 483  3 pg / ml , p < 0.05 ) . as shown in figure 2 , hexanic and ethanolic extracts of t. foenum - graecum , like estradiol , diminished the il-6 level in ovariectomized animals ( p < 0.05 as compared with saline group ) . treatment with 50 and 150 mg / kg of he was also able to significantly decrease serum tnf- from 999  2.4 pg / ml ( saline group ) to 457  2.5 and 445  2.6 pg / ml , respectively , comparable to the effect of estradiol ( 430  2.6 pg / ml ) . a similar decrease in tnf- concentration was induced by both doses of ee ( figure 3 ) . the animals were subjected to ovariectomy surgery and then treated intraperitoneally with 10 g / kg estradiol or concentrations of 50 mg / kg and 150 mg / kg ofhe and ee of t. foenum - graecum seeds for 6 weeks . p < 0.05 vs. control group ; # p < 0.05 vs. saline group . the animals were subjected to ovariectomy surgery and then treated intraperitoneally with 10 g / kg estradiol or concentrations of 50 mg / kg and 150 mg / kg of he and ee of t. foenum - graecum seeds for 6 weeks . * p < 0.05 vs. control group ; # p < 0.05 vs. saline group . the prevalence of metabolic syndrome and inflammatory diseases increases during menopause with the overproduction of proinflammatory cytokines . the present study showed that both he and ee of t. foenum - graecum seed are able to improve metabolic status and decrease level of these cytokines in ovariectomized rats . ovariectomy is a standard experimental model of menopause in rodent to investigate pharmacological management of postmenopausal symptoms ( 11 , 12 , 24 , 25 ) . in our study , in agreement with previous reports , ovariectomy led to increased body weight gain ( 25 , 26 ) . similarly , abbas and elsamanoudy observed that estradiol decreases fbg , fasting insulin , and homeostatis model assessment of insulin resistance ( homa - ir , a predictor of insulin sensitivity ) in ovariectomized rats ( 24 ) . the beneficial effects of estradiol on glucose homeostasis and body weight were mimicked by t. foenum - graecum . hypoglycemic effect of t. foenum - graecum has been demonstrated previously in several experimental and clinical studies ( 27 ) . this effect may be achieved by inhibition of intestinal glucose absorption , enhancing insulin actions on tissues , and activation of hepatic enzymes ( 28 , 29 ) . the inhibitory effect of t. foenum - graecum on weight gain of ovariectomized rats may be related to improvement of insulin resistance . this is in consistent with previous reports that t. foenum - graecum reduces the body weight gain and hyperlipidaemia induced by high - fat diet ( 30 , 31 ) . as reported by other researchers , in our study ovariectomy led to increased level of proinflammatory cytokines . the mechanism underlying elevated inflammation markers in postmenopausal period is not exactly clear ; however , excess production of cytokines by fat and bone marrow can be involved ( 4 , 32 ) . the cytokines tnf- , il-1 , and il-6 have obtained the most attention due to their prominent causal role in postmenopausal inflammatory diseases ( 7 , 33 ) . in this work , estradiol replacement could restore the level of these cytokines to near that of control animals . the mechanisms by which estrogen modulates the level and activity of cytokines may include alterations of immune cell function , antioxidative actions , and changes in nitric oxide activity ( 6 , 32 , 33 ) . similar mechanisms may describe an inhibitory effect of t. foenum - graecum on the level of tnf- , il-1 , and il-6 . it has been reported that this plant has estrogenic activities in vitro ( 34 ) . the presence of phytoestrogen compounds like diosgenin in t. foenum - graecum may be responsible for this estrogenic effect ( 35 ) . on the other hand , several studies have shown that t. foenum - graecum has antioxidative , anti - inflammatory , and immunomodulatory activities in male and female animals ( 16 - 19 ) . for example , it was demonstrated that diosgenin induces anti - inflammatory action in adipose tissue ( 36 ) . reported that ee of t. foenum graecum decreases the blood levels of il-1 , il-2 , il-6 , and tnf- in female rat model of arthritis ( 19 ) also , treatment with hydroalcoholic extract of t. foenum graecum seed reduces oxidative stress and the proinflammatory cytokines tnf- and il-6 in diabetic rats ( 37 ) . further , it has been reported that improvement in glucose metabolism with weight loss is independently associated with reduction of cytokine levels ( 38 ) . in this study , we used ee and he of t. foenum - graecum to test beneficial effects of this plant in the management of metabolic and inflammatory alterations associated with menopause . for extraction of plant materials , nonpolar solvent such as hexane is used to extract lipophilic constituents ( e.g. fatty acids , alkanes , sterols , coumarins , and some terpenoids ) . solvents with medium - polarity such as ethyl acetate are used to extract constituents with intermediate polarity ( e.g. flavonoids and some alkaloids ) . a more polar solvent such as ethanol is used to extract more polar constituents ( e.g. tannins and glycosides ) ( 39 , 40 ) . interestingly , in our experiment , both ee and he of t. foenum - graecum could correct metabolic and inflammatory alterations induced by ovariectomy ( although the he showed stronger effects ) . therefore , it can be concluded that both polar and nonpolar constituents of t. foenum - graecum are involved in the beneficial effects of this plant . diosgenin , a steroid saponin with anti - inflammatory , antioxidative , and estrogenic properties , may be of the candidate constituents responsible for the above - mentioned effects of t. foenum - graecum ( 31 , 35 , 36 ) . further works on isolation and characterization of active compound of the plant would be of interest . this study described observations on the effect of t. foenum - graecum on metabolic and inflammatory alterations in a rat model of menopause , which may not mimic human menopause . another limitation of the present work is the failure to address exact compound responsible for the effects of t. foenum - graecum . also , long - term safety and potential adverse effects of t. foenum - graecum in menopause women should be well - defined . in conclusion , the present study showed that the administration of t. foenum - graecum improves metabolic features , and corrects inflammatory alterations associated with ovariectomy and thus has a potential for management of menopause .