李超,王巍,杨雪卿
LI Chao,WANG Wei,YANG Xueqing

• 1:北京中医医院顺义医院心内科

摘要(Abstract)：

目的:探究泻肺利水合剂对慢性心力衰竭(CHF)大鼠心肌组织氧化应激和线粒体自噬的影响。方法:60只SD大鼠均分为对照组(Control组)、模型组(Model组)及泻肺利水合剂药物处理组(TM组),每组20只; Model组及TM组大鼠给予2. 0 g/L阿霉素以2. 0 m L/kg腹腔注射、1次/周、连续6周; TM组大鼠给予泻肺利水合剂(10 m L/kg·d)灌胃治疗,Control组及Model组大鼠给予等量处理、1次/d、连续6周;灌胃结束时,采用超声心动图检测各组大鼠左室舒张末期内径(LVDD)、左室收缩末期内径(LVSD)、左室缩短分数(LVFS)及左室射血分数(LVEF)的大小;采用酶联免疫吸附试验(ELISA)检测各组大鼠心尖组织中超氧化物歧化酶(SOD)、活性氧(ROS)及丙二醛(MDA)含量,采用Western blot检测各组大鼠心尖组织自噬微管相关蛋白轻链3抗体(LC3-Ⅱ/LC3-Ⅰ)、p62及细胞色素C氧化酶Ⅳ抗体(COXⅣ)蛋白的表达,采用透射电镜观察各组大鼠心尖组织的线粒体自噬小体数量。结果:与Control组相比,Model组和TM组大鼠LVEF和FS水平降低、LVDD和LVSD升高(P <0. 01),Model组心尖组织MDA、ROS、LC3-Ⅱ/LC3-Ⅰ及COXⅣ升高、SOD和p62降低、线粒体自噬数量明显增加(P <0. 05或P <0. 01),TM组心尖组织MDA和ROS升高、SOD降低(P <0. 01);与Model组相比,TM组大鼠LVEF和FS水平升高、LVDD和LVSD降低(P <0. 05或P <0. 01),心尖组织MDA、ROS、LC3-Ⅱ/LC3-Ⅰ及COXⅣ降低、SOD和p62升高、线粒体自噬数量减少(P <0. 05或P <0. 01)。结论:泻肺利水合剂可能通过抑制CHF大鼠心肌氧化应激和线粒体自噬,从而改善其心脏功能。
Objective: To explore the effect of xiefei lishui mixture on oxidative stress and mitochondrial autophagy in the myocardium of rats with chronic heart failure( CHF). Methods: 60 Speleger Dore( SD) rats were randomly divided into CHF Model group( doxorubicin 2. 0 m L/kg intraperitoneal injection,1 time/week for 6 weeks),TM group( Treatment group with xiefei lishui mixture,after doxorubicin 2. 0 m L/kg intraperitoneal injection of Xiefei Lishui Mixture 10 m L/( kg·d) gavage,once a day,for 6 weeks;) and the control group( without modeling,and gastric perfusion of the same amount of normal saline was conducted,once a day,for 6 consecutive weeks). After gastric perfusion,the left ventricular end-diastolic diameter( LVDD),left ventricular end-systolic diameter( LVSD),left ventricular fraction shortening( LVFS),and left ventricular ejection fraction( LVEF) were measured by echocardiography. After successfully modeled rats were given continuous gastric administration for 6 weeks,the rats in each group were anesthetized with 10% chloral hydrate( 4. 5 m L/kg). The chest cavity was opened and the apical tissue was ground into a tissue homogenate,and bicincholin was used( BCA method was used to determine the total protein concentration in the apical tissue of each group of rats,And the enzyme-linked immunosorbent assay( ELISA) was used to detect the contents of superoxide dismutase( SOD),reactive oxygen species( ROS) and malondialdehyde( MDA) in the apical tissues of each group. Western blot was used to detect the expression of autophagy microtubule-associated protein light chain 3 antibody( LC3-Ⅱ/LC3-Ⅰ),p62 and cytochrome C oxidase IV antibody( COX Ⅳ) protein in the apical tissues of each group. Then transmission electron microscopy was used to observe the number of mitochondrial autophagosomes in the apical tissues of rats in each group. Results: Compared with the control group,the levels of LVEF and FS decreased and the LVDD while LVSD increased in Model and TM groups( P < 0. 01),MDA,ROS,LC3-Ⅱ/LC3-Ⅰ,and COX Ⅳ significantly increased,and SOD and p62 significantly decreased,and the number of mitochondrial autophagy increased significantly in apical tissues of TM group( P < 0. 05 or P < 0. 01),and MDA and ROS in apical tissues of TM group increased,while SOD decreased( P < 0. 01); Compared with Model group,the LVEF and FS levels increased and LVDD and LVSD decreased( P < 0. 05 or P < 0. 01),MDA,ROS,LC3-Ⅱ/LC3-Ⅰand COX Ⅳof apical tissues significantly decreased and SOD and p62 increased significantly,and the number of mitochondrial autophagy decreased significantly( P < 0. 05 or P < 0. 01). Conclusion:Xiefei lishui mixture can improve cardiac function in rats with CHF by inhibiting myocardial oxidative stress and mitochondrial autophagy.

关键词(KeyWords)： 大鼠;慢性心衰;泻肺利水合剂;氧化应激;线粒体自噬;心脏功能
rats;chronic heart failure(CHF);xiefei lishui mixture;oxidative stress;mitochondrial autophagy;heart function

Abstract:

Keywords:

基金项目(Foundation): 北京市科技计划课题(Z181100001718072)

作者(Author): 李超,王巍,杨雪卿
LI Chao,WANG Wei,YANG Xueqing

DOI: 10.19367/j.cnki.2096-8388.2020.09.009

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