The Effects Of Class Ⅲ Histone Deacetylase Sirt1on Adipogenic Differentiation Of Mesenchymal Stem Cell C3H10T1/2

Objective: To investigate the roles of class Ⅲ histone deacetylase SIRT1onadipogenic differentiation of C3H10T1/2and the potential molecular mechanism.Methods: An in vitro adipogenesis model was established by using C3H10T1/2，amesenchymal stem cell line. The transcript alternation of SIRT1caused by SIRT1inhibitor EX-527and SIRT11activator resveratrol treatment was detected byRT-PCR. MTT and FACS were adpoted to reveal the effects of EX-527andresveratrol on cell viability and cell cycle respectively. The expression of PPAR-γduring adipogenic differentiation and after SIRT1inhibitor EX-527and SIRT11activator resveratrol treatment were detected by Western blotting.Result：The adipogenic differentiation model of C3H10T1/2cells was successfullyestablished. RT-PCR revealed that resveratrol increased SIRT1transcription, on thecontrary, the SIRT1was suppressed by EX-527; C3H10T1/2cells treated with ADMand resveratrol which is SIRT1activator changed their adipogenic differentiationability. Resveratrol inhibited the adipogenic differentiation of C3H10T1/2cells in adose-dependent manner; EX-527promoted adipogenic differentiation of C3H10T1/2cells in a dose-dependent manner. Cells treated with EX-527arrested their cell cycleat the G0/G1phase and changed their multiangle shape to flat. However, resveratroldoes not obviously influence on cell cycle of C3H10T1/2. The mRNA expression ofadipogenic differentiation key transcription factors PPAR-γ, adipogenic markersdecreased by adding resveratrol in ADM to treat C3H10T1/2.But EX-527promotedthe mRNA expression of adipogenic differentiation key transcription factors PPAR-γ,adipogenic markers.With adipogenic differentiation of C3H10T1/2proceeding,expression level of PPAR-γ mRNA increased. Compared with ADM group.Resveratrol treating group reduces expression level of PPAR-γ mRNA duringadipogenic differentiation of C3H10T1/2cells; Cells disposed with EX-527enhancedexpression level of PPAR-γ mRNA.Conclusion: These data suggested that Class Ⅲ histone deacetylase SIRT1may regulate adipogenic differentiation of C3H10T1/2by down-regulating the expressionof PPAR-γ, a key transcription factors of adipogenic differentiation.