黄芪多糖通过调节Drp1介导的线粒体分裂改善感染性心肌病  

Amelioration of mice septic cardiomyopathy after treatment with Astragalus polysaccharide via regulating mitochondrial fission mediated by Drp1

作　　者：陈旭红[1,3] 纪桢 苑雅茹[1] 边希云 刘晓智[2,3] 袁俐[1,3] Chen Xuhong;Ji Zhen;Yuan Yaru;Bian Xiyun;Liu Xiaozhi;Yuan Li(Department of Obstetrics and Gynecology,the Fifth Central Hospital of Tianjin,Tianjin 300450,China;Central Laboratory,the Fifth Central Hospital of Tianjin,Tianjin 300450,China;Tianjin Key Laboratory of Epigenetics for Organ Development of Premature Infants,the Fifth Central Hospital of Tianjin,Tianjin 300450,China)

机构地区：[1]天津市第五中心医院妇产科,天津300450 [2]天津市第五中心医院中心实验室,天津300450 [3]天津市第五中心医院天津市早产儿器官发育表观遗传重点实验室,天津300450

出　　处：《中国中西医结合急救杂志》2021年第4期455-460,共6页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：天津市滨海新区卫生和计划生育委员会科技项目(2018BWKY013)。

摘　　要：目的从线粒体动力相关蛋白1(Dp1)介导线粒体分裂平衡角度,解读黄芪多糖(APS)对感染性心肌病小鼠的心功能保护作用及机制。方法按随机数字表法将18只成年雄性C57BL/6J小鼠分为对照组、脂多糖(LPS)组和LPS+APS组,每组6只。采用LPS诱导制备感染性:心肌病模型。各组均于术后6h经尾静脉取外周血,采用酶联免疫吸附试验(ELISA)检测小鼠血清炎性因子水平;LPS处理后24 h行超声心动图检查心室功能,包括左室射血分数(LVEF)、左室短轴缩短率(LVFS)、左室舒张期末内径(LVEDD)和左室收缩期末内径(LVESD);检查后即刻处死小鼠取心肌组织,采用蛋白质印迹试验(Western bloting)检测肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、C3a、Drp1、线粒体分裂蛋白1(Fis1)、线粒体融合蛋白2(Mfn2)、视神经菱缩蛋白1(Opa1)蛋白表达。将购买的小鼠心房肌细胞株HL-1分为对照组、LPS组、LPS+APS组和LPS+APS+线粒体氧化磷酸化解偶联剂(FCCP)组。采用活性氧(ROS)检测试剂盒检测分离线粒体及HL-1细胞内的ROS含量:采用线粒体膜电位检测试剂盒(JC-1)检测线粒体膜电位(MMP);采用三磷酸腺背(ATP)检测试剂盒检测HL-1细胞内ATP含量。结果与对照组比较.LPS组小鼠心脏的LVEF、LNFS、LVEDD和LVESD均明显降低.线粒体内ROS含量明显升高,MMP和ATP含量均明显下降;而APS可改善感染性心肌病小鼠的心功能指标[LVEF:0.571±0.026比0.287±0.045.LVFS(%):33.13±2.11比21.22±0.99.LVEDD(mm):3.74±0.10比2.77±0.05.LVESD(mm):2.74±0.09比1.99±0.04.均P<0.05].明显降低心肌内ROS含量[ROS(%):138.39±9.28比260.97±19.22.P<0.05].明显抑制MMP和ATP含量下降[MMP(红色1绿色荧光强度比值):0.91±0.05比0.55±0.01,ATP(%):94.22±10.11比58.74±5.39.均P<0.05),且APS可明显抑制LPS处理后的Drp1和Fis1蛋白过表达以及MIn2和Opal蛋白低表达[Drp1/GAPDH(%):126.33±11.70比218.75±19.44,Fis1/CAPDH(%):105.71±15.77比194.39±5.27,Mfn2/CAPDH(%):92.75±Objective From the point of view of mitoc hondrial fissure balance mediated by mitochondrial dynamic related protein 1(Drp1)to probe the protective efeet and mechanism of Astragalus polysaccharide(APS)on cardiac function of mice with septic cardiomyopathy.Methods Eighteen adul male C57BL/6J mice were randomly divided into 3 groups:control,lipopolysaccharide(LPS)and LPS+APS groups,with 6 mice in each group.LPS was used to induce the mice model of septic eardiomyopathy.After 6 hours,the peripheral blood was laken from the mouse eaudal vein of each group.and the levels of serum inflammatory factors were detected by enzy me-linked immunosorbent assay(ELISA):24 hours after LPS treatment,ventricular function was examined by echocardiogaphy.including left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS).left ventricular end diastolic diameter(LVEDD)and left ventricular end systolie diameter(LVESD);the mice were sacrificed immediately after examinations to obtain cardiac issue.The protein expression levels of lumor nerosis factor-a(TNF-a).interleukin-6(1L-6),C3a,Drpl.mitochondrial mitotie protein I(FisI).mitochondrial fusion protein 2(Mfn2)and optic atrophy protein I(Opal)were detected by W esterm blotting.The purehased mouse atrial muscle cell line HL-1 was divided into 4 groups:control,LPS,LPS+APS and LPS+APS+rarhonyl cyanide 4-trifluoromethoxy)phenylhydrazone(FCCP)groups.Reactive oxygen species(ROS)assay kit was used to detect the content of ROS in isolated mitochondria and HL-I cells:mitochondrial membrane potential(MMP)was detected by mitochondrial membrane potential detection kit(JC-1):the content of ATP in HL-1 rells was deterted by adenosine triphosphate(ATP)detetion kit.Results Compared with those of the control group,the levels of LNEF,LVFS,LNEDD and LVESD were significantly dereased,the rontent of ROS in mitorhondria was signifcantly increased.and the contents of MMIP and ATP were significantly deereaserd in the hearts of LPS mice group;while APS could improve the indexes of car

关 键 词：黄芪多糖 感染性心肌病 线粒体动力相关蛋白1 氧化应激 线粒体分裂 

分 类 号：R54[医药卫生—心血管疾病]