Rho在二氧化硅诱导的人支气管上皮细胞α平滑肌肌动蛋白表达中的作用(英文)

Effect of Rho signaling pathway on SiO_2 induced α-SMA expression in human bronchial epithelial cells

目的:探讨Rho在二氧化硅(SiO2)诱导人支气管上皮细胞(HBECs)α-平滑肌肌动蛋白(α-SMA)表达中的作用。方法:以SiO2处理的HBE细胞为研究对象,免疫细胞化学及Western印迹检测α-SMA表达;GST pull down分析Rho的活化情况;干预实验中用Rho抑制剂Y27632处理细胞后,West-ern印迹检测SiO2对α-SMA表达的影响。结果:SiO2(0,50,100,200,300μg/mL)分别作用HBECs细胞72h后,α-SMA表达增强,其中以200μg/mL SiO2组α-SMA表达最强,为对照组的(5.09±1.98)倍(P<0.01);用200μg/mL SiO2分别作用HBECs细胞1,2,6,12,24h后,Rho明显活化(P<0.01);Rho抑制剂Y27632明显抑制了SiO2诱导的α-SMA表达,20和30μmol/L的Y27632抑制率分别为68%和75%(P<0.01)。结论:Rho参与介导SiO2诱导的HBECs中α-SMA的表达。

Objective To investigate the role of Rho in SiO2 induced α-SMA expression in human bronchial epithelial cells （ HBECs） . Methods HBECs were cultured and stimulated with SiO2. Immunocytochemistry and Western blot were used to detect the expression of α-SMA. The activity of Rho was determined by GST pull down assay. In the prevention experiment,SiO2-stimulated HBECs were incubated with Rho inhibitor Y27632,and the expression of α-SMA was examined by Western blot. Results With SiO2（ 0 - 300 μg/mL） treatment,the expression of α-SMA increased gradually,and 200 μg/mL of SiO2 led to the highest expression of α-SMA which was （ 5. 09 ± 1. 98） times of the expression of α-SMA in the control group（ P 0. 01） . HBECs treated with SiO2（ 200 μg/mL） for indicated time （ 1,2,6,12, and 24 h） showed an obvious increase of Rho activity（ P 0. 01） . Y27632 inhibited SiO2-induced α-SMA expression significantly,and the inhibition rate of 20 and 30 μmol/L Y27632 was 68% and 75% , respectively （ P 0. 01） . Conclusion Rho signaling pathway may mediate SiO2 induced α-SMA expression in HBECs.