摘要
目的:探讨胰岛素对肺泡上皮细胞A549上皮钠通道β亚基(β-ENaC)表达的影响,阐明胰岛素调控β-ENaC的可能信号途径。方法:体外培养肺泡上皮细胞A549,将细胞分为对照组、胰岛素组[分别以不同浓度(2、20和200mU·L-1)胰岛素作用不同时间(30、60和120min)]和LY294002组(胰岛素+LY294002)。采用RT-PCR和Western blotting法测定各组细胞中β-ENaC mRNA和蛋白表达水平以及磷酸化蛋白激酶B(p-Akt)蛋白表达水平。结果:与对照组比较,给予不同浓度(2、20和200mU·L-1)胰岛素作用后,β-ENaC mRNA和蛋白表达水平随浓度增加而增加(P<0.05)。与对照组比较,给予不同浓度(2、20和200mU·L-1)的胰岛素作用后,p-Akt蛋白表达水平随浓度增加而增加(P<0.05)。与对照组比较,胰岛素作用不同时间(30、60和120min)后,β-ENaC mRNA和蛋白表达水平随时间延长而增加(P<0.05)。与胰岛素组比较,LY294002组β-ENaC mRNA和蛋白表达及p-Akt蛋白表达水平降低(P<0.05)。结论:激活PI3K/Akt通路能上调β-ENaC mRNA和蛋白表达水平,有益于肺水肿液的清除。
Objective To investigate the role of insulin in the expression of sodium channel β subunit （β-ENaC） in alveolar epithelial A549 cells, and to clarify the possible signaling pathway on regulating β-ENaC by insulin. Methods The alveolar epithelial A549 cells were cultured in vitro and divided into control group, insulin group （A549 cells were incubated with 2, 20, and 200 mU · L^-1 insulin for 30, 60, and 120 rain）, and LY294002 group, respectively. The expression levels of mRNA and protein of β-ENaC and p-Akt protein were determined by RT-PCR and Western blotting methods. Results Compared with control group, the expression levels of β-ENaC mRNA and protein were elevated significantly after treated with 2, 20, and 200 mU · L^-1 insulin in a concentration-dependent manner（P〈0.05）. Compared with control group, the expression levels of p-AKt protein were elevated significantly after treated with 2, 20, and 200 mU · L^-1 insulin（P〈0.05） in a concentration-dependent manner. Compared with control group, the expression levels of β-ENaC mRNA and protein were elevated significantly after treated with insulin for 30, 60, and 120 min in a time-dependent manner （P〈0.05）. Compared with insulin group, the expression levels of β-ENaC mRNA and protein and p-Akt protein in LY294002 group were decreased （P〈0.05）. Conclosion Activation of PI3K/Akt signaling pathway can increase the expression levels of β-ENaC mRNA and protein, which is beneficial for clearance of pulmonary edema fluid.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2014年第2期233-237,共5页
Journal of Jilin University：Medicine Edition
基金
国家自然科学基金资助课题(81270141)
