强心汤对慢性心力衰竭模型大鼠梗死心肌区组织活性氧及PINK1/parkin介导的线粒体自噬的影响  

Effect of Qiangxin Decoction(强心汤)on Reactive Oxygen Species and PINK1/parkin Mediated Mitochondrial Autophagy in Myocardium of Chronic Heart Failure Model Rats

作　　者：庞延 卢健棋[1] 唐梅玲 黄溥玮 蔡宗余 石炜琦 刘水花 张诗雨 黄敏[2] PANG Yan;LU Jianqi;TANG Meiling;HUANG Puwei;CAI Zongyu;SHI Weiqi;LIU Shuihua;ZHANG Shiyu;HUANG Min(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning,Guangxi Zhuang Autonomous Region,530001;Guangxi University of Chinese Medicine)

机构地区：[1]广西中医药大学第一附属医院,广西壮族自治区南宁市530001 [2]广西中医药大学

出　　处：《中医杂志》2023年第7期722-728,共7页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82160887,81673891);广西自然科学基金(2021GXNSFAA220111,2021GXNSFBA196018);国家中医临床研究基地业务建设第二批科研专项课题(JDZX2015146)。

摘　　要：目的探讨强心汤治疗慢性心力衰竭的可能作用机制。方法40只SPF级SD大鼠随机分为假手术组、模型组、强心汤组、卡托普利组、卡托普利+强心汤组,每组8只。除假手术组外其余各组大鼠均采用结扎左冠状动脉前降支方法建立慢性心力衰竭模型。造模成功后第2天开始,强心汤组大鼠给予12.25 g/(kg·d)强心汤混悬液灌胃;卡托普利组大鼠给予10.38 mg/(kg·d)卡托普利片混悬液灌胃;卡托普利+强心汤组给予强心汤和卡托普利片混悬液灌胃,剂量同上;假手术组和模型组大鼠给予2 ml生理盐水灌胃。各组大鼠均每天灌胃2次,连续28天。采用心脏彩超检测各组大鼠心功能指标[包括左室舒张末期内径(LVEDd)、左室收缩末期内径(LVEDs)、左室射血分数(LVEF)];荧光探针染色检测梗死心肌区组织活性氧(ROS)水平;HE染色观察梗死心肌区组织病理学变化;分别采用RT-PCR和蛋白免疫印迹法检测大鼠梗死心肌区组织PTEN诱导激酶1(PINK1)、E3泛素连接酶(parkin)、微管相关蛋白1轻链3(LC3)mRNA表达及PINK1、parkin、微管相关蛋白1轻链3Ⅰ(LC3Ⅰ)、微管相关蛋白1轻链3Ⅱ(LC3Ⅱ)蛋白表达。结果与假手术组比较,模型组大鼠LVEDd、LVEDs升高,LVEF下降,梗死心肌区组织ROS、LC3 mRNA及LC3Ⅰ/LC3Ⅱ值升高,PINK1、parkin mRNA及蛋白表达下降(P<0.05),且心肌组织中大量弥漫性出血,出血灶周围心肌细胞核形态发生变化,部分细胞核发生核碎裂、核溶解,大量炎性细胞浸润。与模型组比较,各药物干预组大鼠上述指标均明显改善(P<0.05);与强心汤组、卡托普利组比较,卡托普利+强心汤组上述指标改善程度更佳(P<0.05);强心汤组与卡托普利组各指标比较差异均无统计学意义(P>0.05)。结论强心汤可能通过上调PINK1、parkin表达促进线粒体自噬而减少ROS和心肌细胞凋亡从而发挥改善心功能的作用,同时强心汤与卡托普利片联合使用可发挥协同作Objective To explore the possible mechanism of Qiangxin Decoction(QXD)in treating chronic heart failure.Methods A total of 40 SD rats of SPF level were randomly divided into sham operation group,model group,QXD group,captopril group,and captopril+QXD group,with eight rats in each group.Except for the sham operation group,the rats were ligated with the anterior descending branch of the left coronary artery to establish the rat model of chronic heart failure.From the second day after successful modeling,each rat in the QXD group was given 12.25 g/kg QXD suspension.Rat in the captopril group was given 10.38 mg/kg captopril suspension by gavage.captopril+QXD group was intervened with QXD and captopril.The rest groups were given 2 ml normal saline by gavage,and each group was cupped twice a day.After 28 consecutive days of gavage,color Doppler echocardiography was performed to test left ventricular end-diastolic diameter(LVEDd),left ventricular end-systolic diameter(LVEDs),and left ventricular ejection fraction(LVEF).The expression of PINK1,parkin,LC3 mRNA and protein in the infarcted myocardium of rats in each group were detected by RT-PCR and Western blot,and HE stain,ROS fluorescent probe staining was used.Results Compared with the sham operation group,LVEDd and LVEDs in the model group increased,LVEF decreased,ROS,LC3II/I protein expression ratio in myocardial tissue increased signi-ficantly,PINK1,parkin mRNA and protein expression decreased significantly(P<0.05).And there was a large amount of diffuse hemorrhage in myocardial tissue,with morphological changes in cardiomyocyte nuclei around the hemorrhage,nuclear fragmentation and nucleolysis in certain nuclei,and infiltration of a large quantity of inflammatory cells.Compared with the model group,the above indexes of rats in each intervention group were significantly improved(P<0.05).Compared with QXD group and captopril group,the improvement of above indicators in captopril+QXD group was more remarkable(P<0.05).There was no significant difference in all indicato

关 键 词：慢性心力衰竭 强心汤 线粒体自噬 活性氧 PTEN诱导激酶1 E3泛素连接酶 卡托普利 

分 类 号：R285.5[医药卫生—中药学]