【摘要】 目的探讨辛伐他汀对体外诱导培养的大鼠骨髓基质干细胞(BMSCs)成骨分化的作用,以及对大鼠BMSCs基因表达谱的影响。方法取6周龄雄性SD大鼠股骨、胫骨BMSCs进行培养,第3天改用成骨诱导培养基,同时实验组(SIM)加入辛伐他汀(1×10-7mol/L),对照组(V)加入等量溶剂。细胞培养的第14天进行碱性磷酸酶(ALP)比活性测定;第21天进行如下检测:Von Kossa染色观察细胞外基质矿化情况;提取、纯化mRNA,逆转录合成cDNA,荧光标记后与大鼠全基因组寡核苷酸芯片(G4130A)杂交,扫描后筛选出差异表达的基因,Real-time RT-PCR验证部分差异表达基因mRNA的表达。结果 (1)SIM组ALP比活性及细胞外基质矿化能力明显强于V组(P<0.05);(2)在22 575个基因中,共检测出678个差异表达基因,其中包括ALP1、TGFβ1、OCN、DLX5、Axin2、BMP-2、IBSP、MMP13等与成骨分化相关的基因。结论辛伐他汀能够促进体外诱导培养的BMSCs向成骨细胞分化,其作用机制可能与调控多种成骨相关基因的表达有关。更多还原

【Abstract】 Objective To investigate the effects of simvastatin on osteoblast differentiated by rat bone marrow stromal cells(BMSCs),as well as the differently expressed genes associated with osteogenesis of the BMSCs during this process.Methods Bone marrow stromal cells from the femurs and tibias of 6-week old rats were cultured,three days later the cells were treated with simvastatin(1×10-7mol/L)(group SIM) or vehicle(group V).Alkaline phosphatase activity was assessed at 14th day,the following assays were performed at 21th day: Von Kossa staining for observing extracellular matrix mineralization;total RNA was extracted for detecting the gene expression profiles by oligonucleotides microarray chip and were confirmed by real-time quantitative RT-PCR.Results(1)Cells in SIM showed significantly higher ALP activity and ability of extracellular matrix mineralization compared with those of group V(P<0.05).(2)Oligonucleotides microarray chip analysis showed that 678 genes out of 22 575 rat genes had differential expression(≥2 fold or ≤0.5 fold),including genes known to be related to osteogenesis,such as ALP1,TGFβ1,OCN,DLX5,Axin2,BMP-2,IBSP,MMP13,etc.Conclusion Simvastatin could promote the osteo-genic differentiation of BMSCs in vitro and many genes associated with osteogenesis may be involved in this process.更多还原