摘要
目的探究AMP活化的蛋白质激酶(AMPK)/过氧化物酶体增殖物激活受体γ辅助因子1-α(PGC-1α)通路在尿毒症心肌病(UCM)大鼠心脏损伤及线粒体能量代谢中的调控机制。方法将30只大鼠随机分为3组:对照组、UCM组、盐酸阿霉素(DOX)组,每组10只。超声心动图检测心功能指标:左室射血分数(LVEF)、左心室舒张末期内径(LVIDD)、E/A,HE染色检测心肌组织病理变化,Masson染色检测心肌组织纤维化水平,Western blotting检测心肌组织AMPK、PGC-1α、磷酸化AMPK(p-AMPK)、磷酸化PGC-1α(p-PGC-1α)蛋白水平,JC-1试剂盒检测线粒体膜电位,线粒体呼吸链复合物Ⅰ~Ⅴ试剂盒检测线粒体复合物Ⅰ~Ⅴ活性,ATP含量检测试剂盒检测ATP水平,透射电子显微镜检测心肌线粒体超微结构。结果与对照组相比,UCM组大鼠心肌纤维紊乱、断裂,心肌间隙可见胶原大量沉积,大鼠LVEF无显著性差异,LVIDD增加,E/A降低(P<0.05),心肌组织中AMPK、PGC-1α表达无显著性差异,p-AMPK、p-PGC-1α蛋白表达增加(P<0.05),心肌线粒体肿胀、空泡化,且部分线粒体嵴消失,线粒体膜电位减少,ATP含量减少(P<0.05)心肌线粒体呼吸链复合物Ⅰ~Ⅴ活性均减少(P<0.05);与UCM组相比,DOX组大鼠心肌纤维损伤改善,心肌间隙胶原沉积减少,LVEF无显著性差异,LVIDD减少,E/A增加(P<0.05),心肌组织中AMPK、PGC-1α表达无显著性差异,p-AMPK、p-PGC-1α蛋白表达减少(P<0.05),心肌线粒体结构改善,线粒体膜电位增加,ATP含量增加(P<0.05),心肌线粒体呼吸链复合物Ⅰ~Ⅴ活性均增加(P<0.05)。结论抑制AMPK/PGC-1α通路磷酸化可改善UCM大鼠心功能及心肌组织病理学变化,改善线粒体能量代谢。
Objective To investigate the regulatory mechanism of AMP-activated protein kinase(AMPK)/peroxisome proliferator-activated receptor gamma coactivatorⅠalpha(PGC-1α)pathway on heart injury and mitochondrial energy metabolism in rats with uremic cardiomyopathy(UCM).Methods Thirty rats were randomly divided into three groups:control group,UCM group and doxorubicin(DOX)group,with 10 rats in each group.Left ventricular ejection fraction(LVEF),left ventricular internal-diastolic dimension(LVIDD)and E/A were measured by echocardiography.HE staining was used to detect the pathological changes of myocardial tissue.Masson staining was used to detect myocardial fibrosis.Western blotting was used to detect the protein levels of AMPK,PGC-1α,phosphorylated AMPK(p-AMPK)and p-PGC-1αin myocardial tissue.Mitochondrial membrane potential was measured by JC-1 kit.The activity of mitochondrial respiratory chain complexⅠ~Ⅴwas detected by mitochondrial respiratory chain complexⅠ~Ⅴkit.ATP level was detected by ATP content detection kit.The ultrastructure of myocardial mitochondria was detected by transmission electron microscope.Results Compared with the control group,the myocardium fibers in UCM group were disorganized and broken,and a large amount of collagen could be seen in the myocardium space.There was no significant difference in LVEF,LVIDD increased,E/A decreased(P<0.05),there was no significant difference in the expression of AMPK and PGC-1αin myocardial tissue,while the expression of p-AMPK and p-PGC-1αincreased(P<0.05),myocardial mitochondria were swollen and vacuolated,some mitochondrial cristae disappeared,mitochondrial membrane potential decreased,ATP content decreased(P<0.05),the activities of myocardial mitochondrial respiratory chain complexⅠ~Ⅴwere decreased(P<0.05).Compared with UCM group,myocardial fiber injury was improved,collagen deposition in myocardial space was reduced,LVEF was not significantly different,LVIDD was decreased,E/A was increased(P<0.05),there was no significant difference in the expression of AMPK and PGC-1αin myocardial tissue,and the expression of p-AMPK and p-PGC-1αdecreased(P<0.05),myocardial mitochondrial structure improved,mitochondrial membrane potential increased,ATP content increased(P<0.05),the activities of myocardial mitochondrial respiratory chain complexⅠ~Ⅴwere increased(P<0.05)in DOX group.Conclusion Inhibition of AMPK/PGC-1αpathway phosphorylation can improve cardiac function and myocardial histopathological changes in UCM rats,improve mitochondria and explore mitochondrial energy metabolism.
作者
倪倩
颜开萍
李淑娟
徐敏
顾佳雨
黄晓琴
袁乙
NI Qian;YAN Kaiping;LI Shujuan;XU Min;GU Jiayu;HUANG Xiaoqin;YUAN Yi(Department of Nephrology,Yancheng No.1 People’s Hospital,Yancheng First Hospital,Affiliated of Nanjing University Medical College,The Fourth Affiliated Hospital of Nantong University,The First Affiliated Hospital of Jiangsu Vocational College of Medicine,Yancheng 224000,Jiangsu,China)
出处
《心血管病学进展》
CAS
2024年第12期1138-1142,共5页
Advances in Cardiovascular Diseases
基金
江苏省卫生健康委员会科研项目(2020113795)。