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  

Coordinated Regulation of the Proliferation of Airway Smooth Muscle Cells by ERK and PI3K Signal Pathways in Asthmatic Rats

作　　者：徐慧[1] 戴元荣[1] 付玉茹 夏梦玲[1] 

机构地区：[1]温州医科大学附属第二医院呼吸科,浙江省温州市325027

出　　处：《中国全科医学》2015年第9期1032-1036,共5页Chinese General Practice

基　　金：浙江省自然科学基金资助项目(Y2080466);温州市科技局项目(Y20130351)

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- β1group,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- β1group 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- β1group,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- β1group significantly higher than the asthma group,and the TGF- β1＋ wortmannin group significantly lower than the TGF- β1group（ 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- β1group significantly higher than the asthma group,and the TGF- β1＋ PD98059 group significantly lower than the TGF- β1group（ P〈0. 01）. Conclusion The signal pathways of PI3 K and ERK may coordinately regulate the proliferation of ASMC stimulated by TGF- β1.

关 键 词：哮喘 信号通路 肌细胞 平滑肌 细胞增殖 磷脂酰肌醇3-激酶类 细胞外调节蛋白激酶 

分 类 号：R256.12[医药卫生—中医内科学]