辛伐他汀对人骨髓基质干细胞向成骨细胞分化过程的影响

目的：观察辛伐他汀（SIM）对体外人骨髓基质干细胞（hBMSCs）向成骨分化的影响,探讨其刺激成骨的作用机制.方法：取健康成人骨髓进行体外成骨诱导培养,实验分为对照组（G1）：不给予药物干预;实验组（G2）：传代后加入SIM1×10-7mol/L.于传代后7,14,21d采用RealtimeRT-PCR检测mRNA水平和BMP-2,Smad1,β-catenin,Frizzled2的表达,并用免疫组织化学检测蛋白水平β-catenin的表达.传代后7d行细胞碱性磷酸酶（ALP）染色和比活性检测;传代后21d行vonKossa染色,检测细胞外基质矿化.结果：SIM作用后7,14,21d3个不同时间点,BMP-2表达均显著高于对照组;Smad1在成骨诱导14,21d时表达高于G1,其余组间差异均不显著;β-catenin免疫组织化学染色2组差异不显著;G2组ALP表达和矿化能力均显著高于G1组.结论：SIM1×10^-7mol/L可促进体外成骨诱导培养的hBMSCs向成骨细胞分化,上调TGF-β/BMPs信号通路部分信号分子表达,但未能显著改变Wnt/β-catenin信号通路部分相关因子的表达.

Effects of simvastatin on differentiation of osteoblasts derived from human bone marrow stromal cells

AIM：To investigate the effects of simvastatin（SIM）on differentiation of osteoblast derived from the cultured human bone marrow stromal cells（hBMSCs）.METHODS：hBMSCs derivedfrom human were cultured in vitro and divided into 2 groups：Control group（G1）,SIM group（G2）.After subculturing,the medium of treatment group（G2）were added 10-7M simvastatin.Real time RT-PCR was performed to evaluate the mRNA expressionsof BMP-2,Smad1,β-catenin,LRP5,Frizzled2 at day 7,14 and 21 after subculturing;The protein level expression of β-catenin was assessed by immuhistochemistry staining;Alkaline phosphatase staining was used to assess osteoblast differentiation at day 7 after subculturing;Von Kossa staining was used to assess extracellular matrix mineralization and at day 21 after subculturing.RESULTS：The expression levels of mRNA of BMP2 in group G2 were significantly higher than those of group G1 at 3 time points;The expression levels of mRNA of Smad1 in group G2 were significantly higher than those of group G1 at 14 and 21 d subculturing,and the mRNA expression levels of other detected genes showed no significant difference between two groups at each time point.Immuhistochemistry staining：No significant difference was found of the protein expression levels of β-catenin at the 3 time points between the 2 groups.the expression level of ALP and the capability of extracellular matrix mineralization in G2 group was significantlyhigher than that of G1 group.CONCLUSION：Treatment with simvastatin（1×10-7mol/L）could promote osteoblastogenesis of HBMSCs in vitro,probably partially from activing TGF-β/BMPs signaling pathway and upregulating expression of its signaling moleculars.No change was found of expression levels of the detected genes of Wnt/β-catenin signaling pathway after simvastatin treatment,further study should be performed to detect the role of Wnt/β-catenin signaling pathway in osteoblastogenesis-promoting effect of simvastatin on hBMSCs.