重组人膜联蛋白A5对内毒素诱发心肌细胞损伤时P-PKCα和P120-catenin表达的影响  

Effect of recombinant human annexin A5 on expression of p-PKCct and p120-catenin during endotoxin-induced damage to cardiomyocytes

作　　者：赵涛[1] 谷长平[2] 刘孟洁[2] 王栋[2] 周飞[2] 龚妍竹 王月兰[1] 

机构地区：[1]日照市人民医院麻醉科,276826 [2]山东省千佛山医院麻醉科,济南市250014

出　　处：《中华麻醉学杂志》2016年第5期610-612,共3页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（81270127,81570074）

摘　　要：目的评价重组人膜联蛋白A5对内毒素诱发心肌细胞损伤时磷酸化蛋白激酶Cα（p-PKCα）和p120-catenin表达的影响。方法体外培养H9c2心肌细胞，采用随机数字表法分为3组（n＝18）：对照组（C组）、内毒素组（L组）和重组人膜联蛋白A5组（A组）。A组加入重组人膜联蛋白A5（终浓度5 ng/ml）孵育2 h，然后L组和A组给予内毒素（终浓度1 μg/ml）孵育。于孵育4 h时，采用凋亡试剂盒检测细胞凋亡率，免疫共聚焦法测定细胞间隙，采用estern blot法测定p-PKCα和p120-catenin的表达。结果与C组比较，L组细胞凋亡率升高，细胞间隙增加，p120-catenin表达下调，p-PKCα表达上调（P〈0.05）；与L组比较，A组细胞凋亡率降低，细胞间隙减少，p120-catenin表达上调，p-PKCα表达下调（P〈0.05）。结论重组人膜联蛋白A5可抑制PKCα磷酸化，进而上调p120-catenin表达，减轻内毒素诱发的心肌细胞损伤。Objective To evaluate the effect of recombinant human annexin A5 on the expression of phosphorylated protein kinase C alpha （p-PKCα） and pl20-catenin during endotoxin-indueed damage to cardiomyocytes. Methods H9c2 ceils cultured in vitro were randomly divided into 3 groups （n = 18 each） using a random number table： control group （ group C） , endotoxin group （ group L） , and recombinant human annexin A5 group （group A）. Recombinant human annexin A5 （final concentration 5 ng/ml） was added, and the cells were incubated for 2 h in group A, and then ]ipopolysaccharide （final concentration 1ug/ml） was added, and the cells were incubated for 4 h in L and A groups. At 4 h of incubation, cell apoptosis was detected using the cell apoptosis detection kit, the intercellular space was measured using the confocal microscopy, and the expression of p-PKCct and pl20-catenin was determined by Western blot. The apoptosis rate was calculated. Results Compared with group C, the apoptosis rate was significantly increased, the intercellular space was significantly widened, the expression of pl20-catenin was significantly down-regulated, and the expression of p-PKCα was significantly up-regulated in group L （P〈0.05）. Compared with group L, the apoptosis rate and intercellular space were significantly decreased, the expression of pl20-catenin was significantly up-regulated, and the expression of p-PKCα was significantly down-regulated in group A （P〈0.05）. Conclusion Recombinant human annexin A5 can inhibit phosphorylation of PKCαand up-regulate the expression of pl20-eatenin, thus attenuating endotoxin-induced damage fo cardiomyccytes.

关 键 词：膜联蛋白A5 内毒素类 肌细胞 心脏 蛋白激酶C 连接蛋白类 

分 类 号：R614[医药卫生—麻醉学]