LL 37 stimulates the functions of adipose derived stromal/stem cells via early growth response 1 and the MAPK pathway
BackgroundLL 37 is a naturally occurring antimicrobial peptide found in the wound bed and assists wound repair. No published study has characterized the role of LL 37 in the function(s) of human mesenchymal stem cells (MSCs) This study investigated the functions of adipose derived stromal/stem cells (ASCs) activated by LL 37 by performing both in vitro assays with cultured cells and in vivo assays with C57BL/6 mice with hair loss.MethodsHuman ASCs were isolated from healthy donors with written informed consent. To examine the effects of LL 37 on ASC function, cell proliferation and migration were measured by a cell counting kit (CCK 8) and a Transwell migration assay. Early growth response 1 (EGR1) mRNA expression was determined by microarray and real time PCR analyses. The protein levels of EGR1 and regenerative factors were analyzed by specific enzyme linked immunosorbent assays and western blotting.ResultsLL 37 treatment enhanced the proliferation and migration of human ASCs expressing formyl peptide receptor like 1. Microarray and real time PCR data showed that EGR1 expression was rapidly and significantly increased by LL 37 treatment. LL 37 treatment also enhanced the production of EGR1. Moreover, small interfering RNA mediated knockdown of EGR1 inhibited LL 37 enhanced ASC proliferation and migration. Activation of mitogen activated protein kinases (MAPKs) was essential not only for LL 37 enhanced ASC proliferation and migration but also EGR1 expression; treatment with a specific inhibitor of extracellular signal regulated kinase, p38, or c Jun N terminal kinase blocked the stimulatory effect of LL 37. EGR1 has a strong paracrine capability and can influence angiogenic factors in ASCs; therefore, we evaluated the secretion levels of vascular endothelial growth factor, thymosin beta 4, monocyte chemoattractant protein 1, and stromal cell derived factor 1. LL 37 treatment increased the secretion of these regenerative factors. Moreover, treatment with the conditioned medium of ASCs pre activated with LL 37 strongly promoted hair growth in vivo.ConclusionsThese findings show that LL 37 increases EGR1 expression and MAPK activation, and that preconditioning of ASCs with LL 37 has a strong potential to promote hair growth in vivo. This study correlates LL 37 with MSC functions (specifically those of ASCs) including cell expansion, cell migration, and paracrine actions, which may be useful in terms of implantation for tissue regeneration.
Stem Cell Research Therapy