SAMP6小鼠骨髓间充质干细胞线粒体呼吸链复合体功能研究

Function of mitochondrial respiration chain complex in bone marrow mesenchymal stem cells of SAMP6 mice

目的:研究自发性老年性骨质疏松模型SAMP6小鼠骨髓间充质干细胞(BMMSCs)的衰老状态、分化功能及线粒体呼吸链复合体活性。方法:分离培养SAMP6及其对照品系SAMR1小鼠BMMSCs,分别以实时定量PCR(RT-PCR)、Western Blot及细胞染色的方法,检测衰老、分化相关指标,并分别检测线粒体呼吸链复合体活性及复合体相关基因的表达。结果:与对照品系SAMR1小鼠相比,SAMP6小鼠BMMSCs的衰老相关基因、蛋白水平及β-gal染色阳性率显著升高(P<0.05);成骨分化相关基因、蛋白水平及茜素红染色阳性率显著降低(P<0.05);线粒体呼吸链复合体Ⅰ和Ⅲ活性显著降低,且呼吸链复合体相关基因表达亦降低(P<0.05)。结论:SAMP6小鼠BMMSCs存在细胞衰老及成骨分化功能障碍,其线粒体呼吸链复合体功能显著下降。

AIM： To detect senescence condition,osteogenic differentiation ability and the activity of mitochondrial respiration chain complex of bone marrow mesenchymal stem cells（ BMMSCs） from senescence accelerated mouse strain prone 6（ SAMP6）. METHODS： BMMSCs of SAMP6 and its control strain SAMR1 mice were isolated and cultured. The cell senescence and osteogenic differentiation ability were detected by RT- PCR,Western Blot and cell staining respectively. The function of mitochondrial respiration complex was examined by activity assay and related gene expression assay. RESULTS： Compared with control strain SAMR1,BMMSCs of SAMP6 performed increase of cellular senescence related genes and protein level,increase of β-gal positive cell ratio（ P〈0. 05）,decline of osteogenic function（ P〈0. 05）,and low activity of respiration chain complex. CONCLUSION： BMMSCs from SAMP6 perform senescence and osteogenic differentiation dysfunction with significant decline of mitochondrial respiration chain complex.