目的 观察缺血后处理(ischemia postconditioning,IPostC)对动脉粥样硬化(Apoe-/-+AS)模型小鼠心肌缺血再灌注(ischemia-reperfusion,I/R)损伤的影响,研究AAV9-HIF-1α对IPostC作用的影响机制.方法 将60只Apoe-/-+AS模型小鼠按照随机数字表法分为6组,即对照组(AS-sham组)、心肌缺血再灌注组(AS-I/R组)、缺血后处理组(AS-IPostC组)、AAV9-HIF-1α+缺血后处理组(AS-AAV9-HIF-1α+IPostC 组)、AAV9-NC+缺血后处理组(AS-AAV9-NC+IPostC组)、AAV9-HIF-1 α+HIF-1 α 抑制剂+缺血后处理组(AS-AAV9-HIF-1α+2ME2+IPostC 组),每组各 6 只.建立心肌I/R模型和IPostC模型;通过尾静脉分别注射腺相关病毒-9与人巨细胞病毒启动子阴性对照(adeno-associated virus-9 with human cytomegalovirus promoter negative control,AAV9-NC)、腺相关病毒 9 与表达 HIF-1α 的人巨细胞病毒启动子(AAV9-HIF-1α).观察心肌组织病理学变化并测定活性氧簇(reactive oxygen species,ROS)、三磷酸腺苷(adenosine triphosphate,ATP)、线粒体呼吸酶活性;Western blot 法检测缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)、ATP 合酶亚基 b(ATP synth ase subunit b,ATP5F1)、腺苷酸转运(adenylate transporter,ANT)蛋白.结果 与AS-sham组比较,AS-I/R组心肌组织损伤严重,ROS含量升高,ATP含量显著降低;与AS-I/R组比较,AS-IPostC组ROS含量降低,ATP含量升高,但线粒体呼吸酶活性仍然较低,而AS-AAV9-HIF-1α+IPostC组心肌损伤明显下降,线粒体呼吸功能显著好转.结论 动脉粥样硬化小鼠心肌I/R损伤时,氧化应激激活致氧自由基释放、线粒体呼吸功能减弱、细胞能量减少、心肌损伤,而AAV9-HIF-1α+IPostC通过上述机制在一定程度上能增强心肌线粒体呼吸功能.
Effect of HIF-1α on Mitochondrial Respiratory Function in Atherosclerotic Mice
Objective To observe the effect of ischemia postconditioning(IPostC)on myocardial ischemia-reperfusion(I/R)in-jury in atherosclerosis(Apoe-/-+AS)model mice,and to study the mechanism of the effect of AAV9-HIF-1α on IPostC.Methods Sixty Apoe-/-+AS model mice were randomly divided into 6groups:control group(AS-sham group),Apoe-/-+AS-myocardial ischemia-reperfusion group(AS-I/R group),Apoe-/-+AS-ischemia postconditioning group(AS-IPostC group),Apoe-/-+AS-AAV9-HIF-1 α+ischemia postconditioning group(AS-AAV9-HIF-1 α+IPostC group),Apoe-/-+AS-AAV9-NC+ischemia postconditioning group(AS-AAV9-NC+IPostC group),Apoe-/-+AS-AAV9-HIF-1α+HIF-1α in-hibitor+ischemia postconditioning group(AS-AAV9-HIF-1α+2ME2+IPostC group),6mice in each group.Establish myocardial I/R models and IPostC models;adeno-associated virus-9 with human cytomegalovirus promoter negative control(AAV9-NC)and ad-eno-associated virus-9 with human cytomegalovirus promoter expressing HIF-1α(AAV9-HIF-1α)was separately injected through the tail vein.The histopathological changes of myocardium was observed,reactive oxygen species(ROS),adenosine triphosphate(ATP),and the activities of mitochondrial respiratory enzyme were detected;hypoxia inducible factor-1 α(HIF-lα),ATP synthase subunit b(ATP5F1),adenylate transporter(ANT)were detected by Western blot assay.Results Compared with AS-sham group,myocardial tissue injury was serious,ROS content was increased,ATP content was significantly decreased in AS-I/R group;compared with the AS-I/R group,ROS content was decreased,ATP content was increased in the AS-IPostC group,but mitochondrial respiratory enzyme activity was still lower,while myocardial injury was significantly decreased in the AS-AAV9-HIF-1α+IPostC group,and mi-tochondrial respiratory function was significantly improved.Conclusion During myocardial I/R injury in atherosclerotic mice,oxygen free radical activated by oxidative stress was released,mitochondrial respiratory function was weakened,cell energy was decreased,and myocardial cell was damaged,while AAV9-HIF-1α+IPostC can enhance myocardial mitochondrial respiratory function to a certain ex-tent through the above mechanisms.
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