磷脂酰肌醇3激酶/蛋白激酶B信号转导通路在胰蛋白酶激活中性粒细胞中的作用  

Activation of phosphatidylinositol 3-kinase/protein kinase B induced by trypsin in neutrophils

作　　者：张成[1] 孙颖[2] 葛春林[1] 郭仁宣[1] 

机构地区：[1]中国医科大学附属第一医院外科,辽宁沈阳110001 [2]解放军第201医院妇产科

出　　处：《中国医科大学学报》2007年第1期30-32,37,共4页Journal of China Medical University

基　　金：辽宁省科技厅科技计划基金资助项目(2001225001-17)

摘　　要：目的:探讨胰蛋白酶对中性粒细胞磷脂酰肌醇3激酶/蛋白激酶B(PI3K/PKB)信号转导通路的激活作用及PI3K抑制剂对其的影响。方法:应用Western blot技术检测特定浓度胰蛋白酶(40nmol/L)作用中性粒细胞后及应用PI3K抑制剂Wortmannin和LY294002后的p-PKB活化情况;同时应用ELISA及RT-PCR技术检测细胞培养液及中性粒细胞内TNF-α、IL-1β蛋白和mRNA的变化。结果:40nmol/L胰蛋白酶作用中性粒细胞后p-PKB的水平显著高于对照组(P<0.01);TNF-α和IL-1β蛋白的表达也出现了同样的趋势;预先应用Wortmannin或LY294002后完全阻断了胰蛋白酶对中性粒细胞p-PKB的激活作用,与胰蛋白酶刺激组比较差异显著(P<0.01);TNF-α和IL-1β蛋白及mRNA也随着p-PKB活性的抑制而表达减弱。结论:胰蛋白酶能够激活中性粒细胞内PI3K/PKB信号传导通路,Wortmannin及LY294002可以抑制胰蛋白酶对此通路的激活。Objective： To investigate whether phesphatidylinositol 3-kinase/protein kinase B （PI3K/PKB） signal transduction pathway could be activated rapidly by trypsin in neutrophils and whether LY294002 and wortmannin,PI3K inhibitors,could inhibit such effect of trypsin. Methods：The levels of phosphorylated PKB （p-PKB） and total PKB were examined by Western blotting in neutrophils after stimulation with 40 nmol/L of trypsin. The inhibitory effects of LY294002 and wortmannin on the activation of PKB induced by trypsin were observed. The levels of tumor necrosis factor-or （TNF-α） and interleukin-1β （IL-1β） protein and mRNA in the culture and neutrophils were determined by enzyme-linked immunosorbent assay and reverse transcriptase polymerase chain reacfon,respectively. Results：The level of p-PKB in neutrophils and the levels of TNF-α and IL- 1β protein in the culture after sfmulation with trypsin were significantly higher than those in normal controls （P 〈 0.01 ）. Wortmannin and LY294002 could completely inhibit the activation of p-PKB induced by trypsin in neutrophils. With the inhibition of p-PKB,the levels of TNF-α and IL-1β protein and mRNA decreased. Conclusion：Trypsin could activate PI3K/PKB signal transduction pathway in neutrophils,and this action of trypsin is PI3K-dependent and could be inhibited or blocked by wortmannin and LY294002.

关 键 词：中性粒细胞 胰蛋白酶 磷脂酰肌醇3激酶 信号转导 

分 类 号：Q257[生物学—细胞生物学]