Effects And Molecular Mechanisms Of Panax Notoginseng Saponins On Osteogenic Potential Of Bone Marrow Stromal Cells

Background And ObjectiveAs human life expectancy has been increasing, the aging of the population age structure have become increasingly prominent. Osteoporosis as a major threat to health of human attracted widespread attention. How to better control the appearance of osteoporosis is an important issue. Numerous studies have shown that many traditional Chinese medicine could promote fracture healing and prevente osteoporosis. Our study is intended to investigate the effects and molecular mechanisms of panax notoginseng saponins(PNS) on osteogenic potential of bone marrow stromal cells (BMSCs). It will become the foundation for Prevention and treatment of osteoporosis.methods1. The regulation effects of PNS on osteogenic potential of BMSCs:BMSCs were obtained from the bone shaft of femora of rats. The third subculture BMSCs were cultured with different medium. Control group(C)medium containing 15% fetal calf serum. Induction group(I)C with 0.01Mβ-glycerophosphate,50mg/l ascorbic acid, 10-8M dexamethasone.Induction group with the concentration of 10mg/L,50mg/L,100mg/L,200mg/L PNS(short for 10P,50P,100P,200P).The Proliefration of BMSCs were evaluated via MTT assay.Osteogenesis of Bone Marrow Stromal Cells were determined by alkaline phosphatase activity and alizarin red-S stain. The expressions of Cbfal ALP BSP and PPARγ2 were detected by RP-PCR.2. Molecular mechanisms of PNS on osteogenic potential of BMSCs:The third subculture BMSCs were cultured with different medium. Induction group(Ⅰ),Induction group with 100mg/L PNS(100P),100P with JNK inhibitor SP600125(100P+SP),100P with p38MAPK inhibitor SB203580(100P+SB),100P with ERK1 /2 inhibitor PD98059(100P+PD). Osteogenesis of BMSCs were determined by alkaline phosphatase activity and cell mineralization. The expressions of Cbfal ALP BSP and PPARγ2 were detected by RP-PCR.Results1.The regulation effects of PNS on osteogenic potential of BMSCs:PNS in a concentration-dependent enhance proliferation of BMSCs,activity of alizarin red quantitation.100P is better than other groups.The mRNA expression of Cbfal ALP and BSP show 100P is higher than other groups, while the mRNA expression of PPARγ2 show 100P is lower than other groups.2. Molecular mechanisms of PNS on osteogenic potential of BMSCs:100P and 100P+SP show better than other groups in alizarin red quantitation and alkaline phosphatase activity (P<0.01). And there is no significant difference between 100P and 100P+SP(P>0.05).The mRNA expression of Cbfal ALP and BSP in 100P is higher than other groups,while the mRNA expression of PPARγ2 in 100P is lower than in other groups. The mRNA expression of Cbfal ALP BSP in 100P+SB and 100P+PD is lower than 100P but higher than Induction group.Conelusion1. PNS in a concentration-dependent enhance the osteogenesis and proliferation of BMSCs.100P is better than other grow.2. PNS enhance the osteogenesis in BMSCs through MAPK signaling pathway ERK1 /2 and P38MAPK.