自噬和线粒体辅酶Q对急性脓毒症大鼠心功能的影响  

Effect of autophagy and mitochondrial coenzyme Q on cardiac function of rats with acute sepsis

作　　者：陈胜利[1,2] 黄锦达 曾其毅[1] 贾玉娥[2] 王金华[1] 

机构地区：[1]南方医科大学附属珠江医院儿科,广州510280 [2]东莞市厚街医院儿科

出　　处：《中华实用儿科临床杂志》2014年第23期1831-1835,共5页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金（81272070）

摘　　要：目的 探讨自噬对急性脓毒症大鼠心功能的影响以及线粒体辅酶 Q（MitoQ）是否通过自噬对急性脓毒症大鼠心功能障碍起保护作用。方法 SD雄性大鼠45只，按随机数字表法随机分9组：对照组，脂多糖（LPS）4 h、6 h、12 h组，LPS＋渥曼青霉素（Wortmannin）4 h组，LPS＋MitoQ 4h、6h组，MitoQ 组，Wortmannin组，每组5只，每只大鼠予LPS（10 mg/kg）腹腔注射，LPS注射后1 h采用尾静脉注射分别给Wortmannin（2 mg/kg）、MitoQ（6.5 μmol/kg），LPS组给等量9 g/L盐水注射。BL-420E ＋生物信号采集系统测定左心室最大收缩压（LVSP）、左心室舒张末压（LVEDP）、左心室压力上升/下降最大速率（±dp/dtmax）数据。留取血标本测血清肌酸激酶同工酶（CK-MB）水平。取出心肌组织ELISA法测活性氧（ROS）水平，Western blot法检测微管相关蛋白轻链3（LC3）蛋白的表达，光镜和电镜观察心肌病理改变。结果 LPS注射后6h LVSP、LVEDP、±dp/dtmax与对照组比较差异均有统计学意义（P 〈0.05）；给 LPS后再给 Wortmannin 4h即出现统计学差异（P 〈0.05）；LPS＋MitoQ 6h组与对照组比较差异有统计学意义（P 〈0.05），但与同时间点LPS组比较差异无统计学意义（P 〉0.05）。血清CK-MB及心肌组织ROS水平LPS6h组、LPS＋MitoQ 6h组和LPS＋Wortmannin4 h组均高于对照组（P 〈0.05）。光镜下对照组心肌细胞排列规则，心肌间质无炎性细胞浸润；LPS 6h组、LPS＋MitoQ 6h组和LPS＋Wortmannin4 h组心肌纤维结构紊乱，间质大量炎性细胞浸润。电镜下，给LPS6h后自噬体增多，随着时间延长到12h自噬体无明显增多，给MitoQ后与相应时间点 LPS组比较自噬体数量无差异。Western blot检测发现LPS6h组和LPS＋MitoQ 6h组LC3-Ⅱ蛋白水平均高于对照组（P 〈0.05），但LPS12h组和 6h组比较差异无统计学意义（P 〉0.05）。结论 线粒体靶向抗氧化剂MitoQ对脓毒症大鼠心功能�Objective To investigate the effects of autophagy on cardiac function and to determine whether the mitochondrial coenzyme Q （MitoQ） prevents cardiac dysfunction, mediated by autophagy, in rats with acute sepsis. Methods Forty-five Sprague Dawley （SD） rats were randomly divided into 9 groups （ n = 5, each group） ： control group,4 h lipopolysaccharide（LPS） group,6 h LPS group, 12 h LPS group,4 h LPS ＋ Wortmannin group,4 h LPS ＋ MitoQ group,6 h LPS ＋ MitoQ group, MitoQ group and Wortmannin group. Rats in LPS ＋ Wortmaunin group and LPS ＋ MitoQ group were intraperitoneally given LPS（ 10 mg/kg） and followed by an injection of Wortmannin（2 mg/kg） and MitoQ （6.5 μmol/kg） via tail vein 1 hour later, respectively. Rats in each group were given the same amount of normal sodium in addition to different intervention drugs. The cardiac function parameters were measured by a BL-420E ＋ biosignal collection system. Blood samples from abdominal aorta were taken at each time point, and creatine kinase MB isoenzyme（CK-MB） content was detected by using the velocity method. The content of reactive oxygen species（ROS） in isolated myocardial tissues in rats was measured by enzyme-linked immunoadsorbent assay（ELISA）. The protein expression of microtubule-associated protein 1 light chain 3 （ LC3 ） was detected by Western blot method. The pathological changes of myocardial tissue were observed by light and electronic microscopy. Results Compared with the control group,the left ventricular systolic pressure（LVSP） ,the rate of the rise in left ventricular pressure（ -＋ dp/dt max） were significantly decreased in 6 h LPS group,6 h LPS ＋ MitoQ group and 4 h LPS ＋ Wortmannin group （ P 〈 0. 05 ） , left ven- tricular end-diastolic pressure（LVEDP） was significantly increased in these 3 groups（ P 〈 0.05 ）. The contents of CKMB and ROS in 6 h LPS group,6 h LPS ＋ MitoQ group and 4 h LPS ＋ Wortmannin group were higher than those in the control group（ P 〈 0.0

关 键 词：自噬 线粒体辅酶Q 心功能 脓毒症 大鼠 

分 类 号：R459.7[医药卫生—急诊医学]