CAJ | 학술논문

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22 575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/EBP δ,Cited,Asc|2,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.
배경:신벌타정가상조골형태발생단백2적표체종이촉진골형성,단시구체분자수평적작용궤제상불청초.목적;종기인수평명학신벌타정재체외유도대서골수기질간세포향성골세포분화과정중대성골상관기인표체보적영향.방법:취대서고골、경골골수기질간세포진행체외배양,급여신벌타정혹안위제간예,배양제7천,제취、순화mRNA,반전록합성cDNA,형광표기후여대서전기인조과핵감산심편잡교、소묘후사선출차이표체적기인,분석여성골분화상관기인.제14,21천분별진행감성린산매화개결절천소홍염색.결과여결론:제14천,감성린산매염색양성세포비례실험조명현다우대조조;천소홍염색표명신벌타정능촉진성골세포적광화능력.재22 575개기인중,공검출2배차이표체기인화표체서렬표기(ESTs)103조,기중포괄여세포증식급성골분화상관차이표체기인,여C/EBP δ,Cited,Asc|2,Ptpn16,Wisp2,Tieg등.결과표명,신벌타정능구촉진골수기질간세포향성골세포분화,기작용궤제여종기인수평조공다충성골상관기인적표체유관.
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 labelina was employed and the samples were then hybridized with oligonucleotide chip to screen the different genes,which were utillzed 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/EBP δ,Cited,Ascl2,Ptpnl6,Wisp2,Tieg,etc.Simvastatin could induce osteoblastic differentiation of bone marrow stromal cells,involving in many osteogenetic-related genes.