辛伐他汀作用下大鼠骨髓基质干细胞向成骨细胞分化中相关基因表达谱: 基因芯片的分析(英文)  

Gene expression profile in osteoblastic differentiation of bone marrow stromal cells stimulated by simvastatin Gene chip analysis

作　　者：孟亚强[1] 张柳[1] 田发明[1,2] 韩大成[1] 郑杰[1] 蔡俊[1] 

机构地区：[1]华北煤炭医学院附属医院骨外科,河北省唐山市063000 [2]河北医科大学,河北省石家庄市050017

出　　处：《中国组织工程研究与临床康复》2010年第11期2081-2085,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：the Natural Science Foundation of Hebei Province, No.C2006000580~~

摘　　要：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 enhanced the expression of bone morphogenetic protein-2 （BMP-2）, which plays an anabolic role in bone metabolism and osteoblastic lineage differentiation. However, little is known about the molecular mechanism of simvastatin on regulation of bone marrow stromal cells differentiation. OBJECTIVE： To investigated the effect of simvastatin on osteoblastic differentiation of bone marrow stromal cells based on genetics level. METHODS： Bone marrow stromal cells derived from femur and tibia were cultured in different mediums with simvastatin or vehicle for 7 days. Following extraction and purification, mRNA was reverse-transcripted into cDNA. Fluorescence labeling was employed and the samples were then hybridized with oligonucleotide chip to screen the different genes, which were utilized to analyze osteogenesis-related factors. Alkaline phosphatase and Von Kossa staining were performed at days 14 and 21, respectively. RESULTS AND CONCLUSIONS： At day 14, alkaline phosphatase-positive cells were more in the experimental group than control group. Von Kossa staining demonstrated that simvastatin could promote BMSCs osteoblastic differentiation and mineralization. Comparative analysis showed that 103 genes out of 22 575 rat genes had differential expression （≥ 2 fold or ≤ 0.5 fold）, and some genes were related to cell proliferation and ostoeblastic differentiation, including C/EBPδ, Cited, Ascl2, Ptpn16, Wisp2, Tieg, etc. Simvastatin could induce osteoblastic differentiation of bone marrow stromal cells, involving in many osteogenetic-related genes.

关 键 词：辛代他汀 骨髓基质干细胞 成骨细胞 基因 大鼠 骨组织工程 

分 类 号：R318[医药卫生—生物医学工程]