磷脂酰肌醇3激酶和细胞外调节蛋白激酶信号通路对支气管哮喘大鼠气管平滑肌细胞增殖的协同调控作用

摘要

目的：探讨磷脂酰肌醇3激酶（PI3K）和细胞外调节蛋白激酶（ERK）信号通路对支气管哮喘（简称哮喘）大鼠气管平滑肌细胞（ ASMC）增殖的协同调控作用。方法6~8周龄SPF级雄性SD 大鼠，复制大鼠慢性哮喘模型，离体培养大鼠气管 ASMC，将细胞分为正常组、哮喘组、转化生长因子β1（ TGF-β1）组、TGF-β1＋PD98059组、TGF-β1＋渥曼青霉素（ wortmannin）组、TGF-β1＋PD98059＋wortmannin组。采用CCK-8法检测各组细胞增殖情况及Western blotting法检测磷酸化蛋白激酶B（p-Akt）和磷酸化细胞外调节蛋白激酶1/2（p-ERK1/2）表达情况。结果 CCK-8法检测各组大鼠ASMC OD 值，TGF-β1组高于正常组和哮喘组，TGF-β1＋ PD98059组、TGF-β1＋wortmannin组和TGF-β1＋PD98059＋wortmannin组低于TGF-β1组，TGF-β1＋ PD98059＋wortmannin组低于TGF-β1＋ PD98059组和TGF-β1＋wortmannin组（ P＜0.01）。Western blotting法检测ASMC中p-Akt的表达，哮喘组高于正常组，TGF-β1组高于哮喘组，TGF-β1＋wortmannin组低于TGF-β1组（ P＜0.01）。Western blotting法检测ASMC中p-ERK1/2的表达，哮喘组高于正常组，TGF-β1组高于哮喘组，TGF-β1＋ PD98059组低于TGF-β1组（P＜0.01）。结论 PI3K和ERK信号通路协同调控了TGF-β1刺激哮喘大鼠ASMC增殖过程。%Objective To investigate the coordinated regulation of the proliferation of airway smooth muscle cells (ASMC)by phosphoinositide -3 - kinase( PI3K) and extracellular regulated protein kinases( ERK)signal pathways in asthmatic rats. Methods SPF Sprague Dawley rats of 6 to 9 weeks old were used to establish chronic asthma model. ASMC was cultured in vitro and the cells were divided into normal group,asthma group,TGF-β1 group,TGF-β1 + PD98059 group, TGF-β1 +wortmannin group and TGF-β1 +PD98059+wortmannin group. CCK-8 method was used to detect cell proliferation and Western blotting method was used to detect the expression of p-Akt and p-ERK1/2. Results The OD value of ASMC was determined by CCK-8 method,with the TGF-β1 group and TGF-β1 +PD98059+wortmannin group significantly higher than the normal and asthma group,the TGF-β1 + PD98059 group and TGF-β1 +wortmannin group significantly lower than the TGF-β1 group,and the TGF-β1 + PD98059+wortmannin group significantly lower than the TGF-β1 + PD98059 group and TGF-β1 +wortmannin group ( P <0. 01 ). The expression level of p - Akt in ASMC was determined by Western blotting method,with the asthma group significantly higher than the normal group,the TGF-β1 group significantly higher than the asthma group,and the TGF-β1 +wortmannin group significantly lower than the TGF-β1 group(P<0. 01). The expression level of p-ERK1/2 was also determined by Western blotting method,with the asthma group significantly higher than the normal group,the TGF-β1 group significantly higher than the asthma group,and the TGF-β1 + PD98059 group significantly lower than the TGF-β1 group(P <0. 01). Conclusion The signal pathways of PI3K and ERK may coordinately regulate the proliferation of ASMC stimulated by TGF-β1 .