重庆医学2017，Vol.46Issue(34)：4757-4762,6.DOI:10.3969/j.issn.1671-8348.2017.34.002
过敏毒素C3a在健康人气道上皮间质转化中的作用研究
Study on the role of allergic toxin C3a in epithelial mesenchymal transition of normal human bronchial epitahelium cells
摘要
Abstract
Objective To investigate the role of anaphylatoxin C3a on epithelial mesenchymal transition(MTT) in normal human bronchial epithelium cells and its molecular mechanism.Methods Normal human bronchial epithelium cells BEAS-2B were cultured,and divided into control group,rhC3a stimulation group,rhC3a+ C3a receptor(C3aRA)antagonist group,the morphological changes of cells were observed by microscope;cell proliferation was detected by MTT;the expression of TGF-β1 protein level in cell supernatant was evaluated by ELISA;the expression of C3aR mRNA and EMT related indicators mRNA changes were detected by RT-PCR;the expression of C3aR and Smad2/3,p38 MAPK pathway proteins were detected by Western blot.Results Cell morphology in 30,50 nmol/L rhC3a stimulation group was changed from normal cobblestone like to spindle shape,cell morphology in C3aRA antagonist group had no significant change when compared with the control group.The cell proliferation was reduced in 50 nmol/L rhC3a stimulation group(P=0.047);the levels of TGF-β1,p-Smad2/3,p-p38-MAPK protein were increased(P＜0.05),C3aR mRNA and protein levels were also significantly increased (P＜0.05) in 30 nmol/L rhC3a stimulation group when compared with control group,but the addition of 1 μmol/L C3aRA could reduce their expressions(P＜0.05).30 nmol/L rhC3a could induce the up regulation of α-SMA and N-cadherin mRNA,and decrease the expression of E-cadherin mRNA,adding 1 μmol/L C3aRA can antagonize this process (P＜0.05).Conclusion Anaphylatoxin C3a can induce EMT in normal human bronchial epithelium cells by combining C3aR,its mechanism may be involved in activating Smad2/3 and p38-MAPK pathway.关键词
气道重塑/纤维化/过敏毒素C3a/上皮间质转化Key words
airway remodeling/fibrosis/anaphylatoxin C3a/epithelial mesenchymal transition分类
医药卫生引用本文复制引用
李竹,刘代顺,吴凯峰,丁宏伟,刘建英..过敏毒素C3a在健康人气道上皮间质转化中的作用研究[J].重庆医学,2017,46(34):4757-4762,6.基金项目
贵州省科学技术基金(黔科合J字[2013]2310号). （黔科合J字[2013]2310号）
