The Vary Of β-catenin And PSmad1/5/8during Osteogenic Differentiation Of BMSCs Applied Simvastatin

Objectives Simvastatin, an inhibitor of3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, has been known to reduce cholesterol biosynthesis. Recent experimentalstudies have suggested that simvastatin may also have Osteoinductive effects. But the varyof β-catenin and Smad1/5/8during Osteogenic Differentiation of BMSCs expression inearly stages is not yet clear. In this study, we investigated the stimulatory effect ofsimvastatin on the osteogenic differentiation of Bone marrow mesenchymal stem cells(BMSCs) in early stages. Under conditions of Osteoinductive environment, to explore theExpression of β-catenin and Smad1/5/8during Osteogenic Differentiation of BMSCsapplied simvastatin.Methods Twenty SPF4-week-old female SD rats, all the rats were killed by cervicaldislocation and remove both bilateral femur and tibia under sterile conditions. Whole bonemarrow culture method for primary and passaged cultures and take the second generationof cells were randomly divided into three groups. CM Group: complete medium as anegative control; OM Group: containing10mmol/L β-glycerophosphate,50μg/ml ascorbicacid and10-8mol/L dexamethasone osteogenic induction medium, as a positive control;SIM group: added at a concentration of10-7mol/L simvastatin medium; OM+SIM group:added at a concentration of10-7mol/L simvastatin osteogenic induction medium. Afteradministration of48h, cell proliferation of CM and SIM was measured by MTT analysis.ALP staining of CM and OM and SIM was measured after the administration7d. Proteinexpressions of β-catenin and pSmad1/5/8were detected by Western Blot after theadministration1h,2h,4h,8h,12h,24h,36h and48h.Results1MTT assay: Compared with the control group,10-7mol/L simvastatin had noeffect on cell viability.2ALP staining:7d ALP staining of OM group and OM+SIM groupwere higher than the CM group.7d ALP staining of OM+SIM group were highest(P＜0.05).3Western Blot analysis:①β-catenin: In the OM and OM+SIM groups, expressionof β-catenin levels increased in12h,24h,36h and48h(P＜0.05). In the OM+SIM groupexpression of β-catenin levels increased in12h,24h and36h(P＜0.05).Compared with thedifferent group in different times, OM+SIM group compared with OM expression of β-catenin levels increased in12h and36h(P＜0.05). In the48h, OM compared with CM group, the expression of β-catenin levels increased (P＜0.05).②pSmad1/5/8: In the OMgroups, expression of pSmad1/5/8levels increased in8h,12h,24h and48h(P＜0.05). Inthe OM+SIM groups, expression of pSmad1/5/8levels increased in8h,12h and36h(P＜0.05). Compared with the different group in different times, OM+SIM group comparedwith OM expression of pSmad1/5/8levels increased in8h,12h and48h(P＜0.05).Conclusions Simvastatin of10-7mol/L had no effect on cell viability. Simvastatin havethe ability of inducing osteogenesis. The expression of β-catenin and pSmad1/5/8levelsincreased by simvastatin.