p55 TNFR 选择性淋巴毒素对心肌细胞氧化应激损伤的保护机制研究  

The mechanism of protection against myocardial oxidative stress injury by anovel lymphotoxinbinding p55TNFRselectively

作　　者：苏文利[1] 郝平[1] 朱文献[1] 王艳华[1] 邵伟怡 钱龙杰[1] 王毅鑫[1] 

机构地区：[1]上海中医药大学附属普陀医院急诊外科,上海200062

出　　处：《中国急救医学》2015年第10期929-933,共5页Chinese Journal of Critical Care Medicine

基　　金：上海市卫生计生系统重点专科建设项目（ZK2012A35）

摘　　要：目的：研究p55 TNFR选择性淋巴毒素对心肌细胞氧化应激损伤的保护作用及其机制。方法取Wistar乳鼠制备原代心肌细胞，建立H2 O2致心肌细胞的氧化应激损伤模型，将细胞分为正常组、模型组、rhLTα处理组及rhLTα－Q107 E处理组，流式细胞术检测细胞凋亡，免疫印迹法检测细胞内NF－κB的活化；实时定量RT－PCR法检测抗凋亡分子的转录水平（ Bcl－2、Bcl－X、XIAP）；免疫印迹法检测MnSOD水平；WST法检测CuZn／Mn－SOD活性。结果在体外大鼠心肌细胞的氧化应激损伤模型中，细胞凋亡率明显增高，伴随着Caspase－3活性明显增高；抗凋亡分子Bcl－2、Bcl－xL和XIAP的表达水平均减少；进一步检测MnSOD的蛋白表达水平下降，CuZn／Mn－SOD活性减弱。当加入rhLTα－Q107 E处理后，可逆转上述改变情况，伴随NF－κB的激活。而野生型LTα作用不明显（P＜0．05）。结论 p55TNFR选择性淋巴毒素rhLTα－Q107 E可通过激活NF－κB，上调抗凋亡分子表达和诱导线粒体MnSOD活化，发挥对抗氧化应激保护心肌细胞的作用，且优于野生型的LTα。Objective To investigate the effect of a novel lymphotoxin with selectively binding to p55 tumor necrosis factor receptor （p55TNFR） on myocardial oxidative stress injury , and explore the mechanism.Methods Primary cardiomyocytes were prepared from hearts of neonatalWistar rats , and myocardial oxidative stress injury model was established by hydrogen peroxide （ H2 O2 ） .Then cells were divided into normal group , H2 O2 treatment group, and treatment groups with rhLTαor rhLTα-Q107E. Cell apoptosis was detected by flow cytometry , and the activation of NF -κB was detected by Western blot analysis .Real -time quantitative RT -PCR assay was applied to detect the expression levels of anti-apoptotic molecules （Bcl-2, Bcl-X and XIAP）.The expression and activities of MnSOD or CuZn-SOD were detected by Western blot or WST .Results Increased cell apoptosis and the activity of Caspase-3 was observed in the oxidative stress injury model of myocardial cells .The expression of anti-apoptotic protein Bcl -2, Bcl-xL and XIAP were decreased .The expression level and activity of MnSOD were decreased .However , the treatment of rhLTα-Q107 E, but not wild type LTα, reversed these changes , along with the activation of NF -κB.Conclusion This work demonstrates that rhLTα-Q107 E plays a role on the protection against myocardial oxidative stress injury , through the activation of NF-κB, up -regulating the expression of the anti -apoptotic molecules , activating mitochondrial MnSOD, which was better than the wild type LTα.

关 键 词：淋巴毒素 氧化应激 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]