辛伐他汀对大鼠BMSCs成骨分化及其基因表达谱的影响

 目的 探讨辛伐他汀对体外培养的大鼠骨髓基质干细胞（bone marrow stromal cells, BMSCs）成骨分化的作用，以及在此过程中大鼠BMSCs基因表达谱的变化。方法 取6周龄雄性SD大鼠股骨、胫骨骨髓，进行细胞培养，并于第3天实验组（SIM）加入辛伐他汀（1×10-7 mol·L-1），对照组（V）加入等量溶剂。细胞培养的第14天提取、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，并于同一时间点进行碱性磷酸酶（ALP）染色；第21天行Von Kossa染色观察细胞外基质矿化情况。 结果 ①ALP染色：SIM组ALP阳性细胞比例显著高于V组；②Von Kossa染色：SIM组细胞外基质矿化能力明显强于V组；③在22 575个基因中，共检测出318个差异表达基因，其中包括ALPl、TGFβ1、IBSP、MMP13、VEGF、IL1α、CXCL10、Zfp36、MEPE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，同时伴有多个成骨相关基因表达水平的改变。

Effects of Simvastatin on Osteogenic Differentiation and on Gene Expresssion Profiles of Osteogenic Differentiated Rat Bone Marrow Stromal Cells

OBJECTIVE To investigate the effects of simvastatin on osteogenic differentiated rat bone marrow stromal cells(BMSCs), as well as the differently expressed genes associated with osteogenesis of the BMSCs. METHODS Bone marrow stromal cells from the femurs and tibias of 6-week old rats were cultured in vitro. The cells were treated with simvastatin (1×10-7 mol·L-1)(group SIM) or vehicle(group V) at 3rd day. Total RNA was extracted for detecting the gene expression by oligonucleotides microarray chip at 14th day, and at the same time, alkaline phosphatase staining was performed. At 21st day， Von Kossa staining was applied for observing extracellular matrix mineralization. RESULTS Cells in SIM showed significantly high ALP-positive staining cell rate compared with that of group V. Von Kossa staining indicated a positive effect of simvstatin on extracellular matrix mineralization. Oligonucleotides microarray chip analysis showed that 318 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 ALPl， TGFβ1， IBSP， MMP13， VEGF， IL1α， CXCL10， Zfp36 and MEPE, etc. CONCLUSION Simvastatin could promote the osteogenic differentiation of BMSCs in vitro, with many genes associated with osteogenesis differentially expressed.