p120在机械通气性牵张致小鼠肺泡上皮细胞E-cadherin胞浆转移中的作用  

Role of p120 in mechanical stretch-induced transferring of E-cadherin to cytoplasm in mouse alveolar epithelial cells

作　　者：李冠军[1] 周飞[2] 王月兰[2] 谷长平[2] 

机构地区：[1]山东省交通医院麻醉科,济南市250031 [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》2017年第6期661-665,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金青年基金（81600054）;山东省自然基金（ZR2015HL001）;中华医师协会中青年学术论坛“人福杯”青年麻醉学医师科研基金（220150800004）

摘　　要：【摘要】目的评价p120一catenin蛋白（p120）在机械通气性牵张致小鼠肺泡上皮细胞E—cad—herin胞浆转移中的作用。方法实验IMLE-12细胞以（1．0～1．5）×106个／孔的密度接种于细胞牵张6孔板，采用随机数字表法分为3组（n=12）：对照组（c组）、牵张2h组（CS2组）和牵张4h组（CS4组）。机械通气性牵张参数：牵张频率O．5Hz，牵张强度20％，牵张间歇比1：1，CS，组和CS4组分别牵张2和4h。C组不进行牵张处理。采用Westernblot法检测p120、E—cadherin及磷酸化Src激酶（p-Src）的表达，胞膜和胞浆E—cadherin的表达。实验ⅡMLE-12细胞以（1．0～1．5）×10。个／孔的密度接种于细胞牵张6孔板，采用随机数字表法分为4组（n=6）：对照组（C组）、牵张组（CS组）、p120siRNA转染组（p120siRNA组）和p120siRNA转染＋牵张组（p120siRNA＋cs组）。c组和cs组采用scramblesiRNA转染，24h后cs组细胞行机械通气性牵张；p120siRNA组和p120siRNA＋cs组经p120siRNA转染，24h后p120siRNA＋CS组后行机械通气性牵张。牵张时间2h，参数设置同实验I。各组处理结束后，采用Westernblot法检测胞膜和胞浆E—cadherin的表达。结果实验I与c组比较，CS2组和CS4组p120和E—cadherin表达下调，p-Src表达上调，胞膜E-cadherin表达下调，胞浆E—cadherin表达上调（P〈0．05）；与CS2组比较，CS4组p120和E—cadherin表达下调，p-Src表达上调，胞膜E—cadherin表达下调，胞浆E．cadherin表达上调（P〈0．05）。实验Ⅱ与C组比较，cs组、p120siRNA组、p120siRNA＋CS组胞膜E—cadherin表达下调，胞浆E—cadherin表达上调（P〈0．05）；与CS组或p120siRNA组比较，p120siRNA＋CS组胞膜E．cadherin表达下调，胞浆E—cadherin表达上调（P〈0．05）。结论p120降解可促进机械通气性牵张诱发的小鼠肺泡上皮细胞E—cadherin胞浆转移。Objective To evaluate the role of pl20-catenin protein （p120） in mechanical stretch- induced transferring of E-cadherin to cytoplasm in mouse alveolar epithelial cells. Methods Experiment I Mouse alveolar epithelial cells （MLE-12 cells） were seeded in 6-well cell stretch plates at a density of （1.0-1.5） ×106 cells/well and divided into 3 groups （n= 12 each） using a random number table： control group （ group C） , cyclic stretch for 2 h group （ group CS2 ） and cyclic stretch for 4 h group （ group CS4 ）. The cells underwent 20% cyclic stretch at 0.5 Hz （stretch ： intermittence = 1 ： 1） for 2 and 4 h in CS2 and CS4 groups, respectively. The cells underwent no cyclic stretch in group C. The expression of p120, E-cadherin and phosphorylated Src kinase （p-Sre） and expression of E-cadherin in cytomembrane and cy- toplasma were detected by Western blot. Experiment II MLE-12 cells were seeded in 6-well cell stretch plates at a density of （1.0-1.5）x 106 cells/well and divided into 4 groups （n = 6 each） using a random number table： control group （group C） , cyclic stretch group （group CS） , p120 small interfering RNA （siRNA） transfection group （group p120 siRNA） , and p120 siRNA transfection plus cyclic stretch group （group p120 siRNA＋CS）. The cells were transfected with scramble siRNA in C and CS groups, and 24 h later the cells underwent 20% cyclic stretch for 2 h at 0.5 Hz （stretch ： intermittence = 1 ： 1） in group CS. The cells were transfected with p120 siRNA in p120 siRNA and p120 siRNA＋CS groups, and 24 h lat- er the cells underwent 20% cyclic stretch for 2 h at 0.5 Hz （ stretch ： intermittence = 1 ： 1） in group p120 siRNA＋CS. The expression of E-cadherin in cytomembrane and cytoplasm was detected by Western blot af- ter the end of＂ treatment in each group. Results Experiment ｜ Compared with group C, the expression of p120 and E-cadherin was significantly down-regulated, the expression of p-Src was up-regulated, the ex-

关 键 词：生物力学 上皮细胞 肺泡 连环素类 钙黏着糖蛋白类 

分 类 号：R614[医药卫生—麻醉学]