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背景:辛伐他汀可上调骨形态发生蛋白2的表达从而促进骨形成,但是具体分子水平的作用机制尚不清楚。目的:从基因水平明确辛伐他汀在体外诱导大鼠骨髓基质干细胞向成骨细胞分化过程中对成骨相关基因表达谱的影响。方法:取大鼠股骨、胫骨骨髓基质干细胞进行体外培养,给予辛伐他汀或安慰剂干预,培养第7天,提取、纯化mRNA,反转录合成cDNA,荧光标记后与大鼠全基因组寡核苷酸芯片杂交、扫描后筛选出差异表达的基因,分析与成骨分化相关基因。第14,21天分别进行碱性磷酸酶和钙结节茜素红染色。结果与结论:第14天,碱性磷酸酶染色阳性细胞比例实验组明显多于对照组;茜素红染色表明辛伐他汀能促进成骨细胞的矿化能力。在22575个基因中,共检出2倍差异表达基因和表达序列标记(ESTs)103条,其中包括与细胞增殖及成骨分化相关差异表达基因,如C/EBPδ,Cited,Ascl2,Ptpn16,Wisp2,Tieg等。结果表明,辛伐他汀能够促进骨髓基质干细胞向成骨细胞分化,其作用机制与从基因水平调控多种成骨相关基因的表达有关。
BACKGROUND: Simvastatin can up-regulate the expression of bone morphogenetic protein-2 to promote bone formation. However, the mechanism of action at specific molecular level is not clear yet. OBJECTIVE: To determine the effect of simvastatin on the osteogenesis-related gene expression profile induced by osteoblast differentiation of rat bone marrow stromal stem cells induced by simvastatin in vitro. Methods: Rat femur and tibia bone marrow stromal stem cells were cultured in vitro and were given simvastatin or placebo. On the 7th day after culture, mRNA was extracted and purified, and cDNA was reverse transcribed. After fluorescent labeling, After hybridization, the differentially expressed genes were screened and the genes related to osteogenic differentiation were analyzed. Alkaline phosphatase and calcium nodular alizarin red staining were performed on the 14th and 21st day respectively. RESULTS AND CONCLUSION: On the 14th day, the ratio of alkaline phosphatase positive cells in the experimental group was significantly higher than that in the control group. Alizarin red staining showed that simvastatin could promote the mineralization of osteoblasts. A total of 22,575 genes were screened, of which 102 differentially expressed genes and 103 ESTs were identified, including differentially expressed genes related to cell proliferation and osteogenic differentiation, such as C / EBPδ, Cited, Ascl2, Ptpn16, Wisp2 Tieg et al. The results show that simvastatin can promote the differentiation of bone marrow stromal cells into osteoblasts. The mechanism of simvastatin is related to the regulation of gene expression of many osteogenic genes.