线粒体损伤在脓毒症大鼠心肌细胞凋亡中作用.docVIP

线粒体损伤在脓毒症大鼠心肌细胞凋亡中作用 　　【摘要】目的 探讨线粒体损伤在脓毒症大鼠心肌细胞凋亡中的作用及其可能机制。方法 72只SD大鼠随机（随机数字法）分为2组：（1）阴性对照组18只，腹腔注射无菌生理盐水；（2）脓毒症组，按大肠杆菌内毒素注射后6、12、24 h分为3个亚组，各18只。取心脏组织，光镜和电镜下观察组织病理学和超微结构的变化，TUNEL法原位检测心肌细胞凋亡并计算凋亡指数，免疫印迹法检测核转录因子-κB（NF-κB）p65的表达，黄嘌呤氧化酶法测定线粒体超氧化物歧化酶（SOD）活性，比色法检测线粒体还原型谷胱甘肽（GPx）活性，硫代巴比妥酸法测定线粒体脂质过氧化产物丙二醛（MDA）含量，OxyBlot TMas 蛋白氧化检测试剂盒观察线粒体蛋白氧化。结果 脓毒症大鼠心肌有炎症细胞浸润，细胞水肿及空泡形成，并且在24 h最明显；心肌细胞凋亡指数和核蛋白NF-κB p65活化均显著增加（P 　　【关键词】脓毒症；心肌损伤；细胞凋亡；线粒体；核转录因子-κB 　　Role of damaged mitochondria in cardiocyte apoptosis in septic rats LI Li， YAN Jing， CHEN Chang-qin， GONG Shi-jin， DAI Hai-wen， YU Yi-hua， CAI Guo-long， CHEN Jin， X Qiang-hong. Intensive Care Unit， Zhejiang Hospital， Hangzhou 310013， China 　　Corresponding author： YAN Jing， Email： yanjing2011@ 　　【Abstract】Objective To investigate the role of damaged mitochondria in cardiac cell apoptosis in septic rats and the possible mechanism involved. Methods Seventy-two Sprague-Dawley rats were randomly （random number） divided into negative control group （n=18） and septic group （further divided into three groups as per rats sacrificed 6 h， 12 h， and 24 h after endotoxin injection intra-peritoneally， n=18）.The hearts of rats were taken. The changes of cardiac morphology were observed under light microscope and scanning electron microscope. Cell apoptosis in situ were examined by using terminal transferase-mediated dUTP nick end-labeling assay and nuclear factor-kappa B （NF-κB） activation in myocardium was detected by using Western blotting to estimate myocardial cell apoptosis.Mitochondrial lipid and protein oxidation were measured to assess oxidative stress， and mitochondrial superoxide dismutase （SOD） and glutathione peroxidase （GPx） activities were determined to estimate antioxidant defense. Results Septic induced inflammatory cells infiltration， myocardium degeneration and effusion in a time-dependent manner. A remarkable expansion of capillaries could be observed in the hearts of infected rats at post-challenge of 24 h. Compared with