中文摘要：

目的探讨线粒体功能在自发性高血压大鼠动脉平滑肌细胞中的改变与机制。方法选择正常血压Wistar-Kyoto（WKY）大鼠为正常对照组（n=16）,同周龄自发性高血压Spontaneous Hypertensive Rat（SHR）大鼠为实验组（n=16）。采用组织块贴壁法进行大鼠胸主动脉平滑肌细胞（vascular smooth muscle cell,VSMC）原代培养并用免疫组织化学法鉴定;VSMCs线粒体功能改变采用荧光法和流式细胞术分别检测胞质内ATP、ROS水平和线粒体膜电位;筛选线粒体呼吸链主要亚基ND1、ND2、Cytb8、CO1、ATP8的mRNA,采用实时定量聚合酶链式反应（qRT-PCR）法观察其在各组VSMCs中的表达变化。结果与正常组相比,SHR组大鼠VSMCs细胞线粒体ATP产生量由（60.873±2.750）nmol/L下降至（51.581±3.280）nmol/L（P〈0.01）,胞质内ROS水平明显升高,从（0.752±0.043）至（1.082±0.146）（P〈0.01）。细胞线粒体膜电位改变,由（1.614±0.173）降至（1.401±0.132）（P〈0.01）,提示VSMCs细胞线粒体功能显著受损。其次,对线粒体呼吸链主要亚基检测发现SHR组ND1亚基mRNA表达存在明显下调,与对照相比有显著差异。结论高血压可导致动脉平滑肌细胞线粒体功能受损和氧化应激水平提高,线粒体氧化呼吸链ND1亚基改变可能是在高血压发病中参与血管平滑肌细胞线粒体功能调控的重要环节。

英文摘要：

Objective To investigate the change of mitochondrial function of aortic smooth muscle cells（VSMCs）and mechanism in spontaneous hypertensive rats.Methods A total of 16 healthy spontaneously hypertensive rats（SHR）and corresponding ages of normal rats（WKY）aged 14 weeks were selected.VSMCs were primarily cultured in vitro using tissue block adhering wall method,and the third to fifth generation was used for cell experiment.First we detected the intercellular ATP using fluorescence technique.ROS and mitochondrial membrane potential of VSMCs were tested by flow cytometry.The mRNAs expression of ND1,ND2,Cytb8,ATP8,CO1 in mitochondrial respiratory chain were analyzed for realtime PCR.Results ATP in SHR group showed significantly downward compared to WKY group（P 〈0.01）,and ROS increased markedly in contrast to WKY group（P 〈0.01）,while the mitochondrial membrane potential decreased significantly in SHR group.Real-time PCR confirmed that ND1 expression in SHR group downregulated strikely（P 〈0.01）,but ND2,Cytb8,ATP8,CO1 expression in SHR group showed no statistically difference compared to WKY group.Conclusion Hypertension would result in mitochondria dysfunction and enhanced level of oxidative stress of VSMCs.The changes of ND1 subunit in the oxidative respiratory chain of VSMCs might play an important part in the pathological genesis of essential hypertension.