摘要
目的 探讨c-Jun氨基末端激酶(JNK)-c-Jun/活化蛋白(AP)-1信号转导通路在血管紧张素Ⅱ(AngⅡ)诱导的肾小球系膜(MC)细胞转化生长因子β1(TGF-β1)表达中的调控作用。方法 应用核酸酶保护法检测系膜细胞TGF-β1 mRNA表达。应用凝胶电泳迁移率(EMSA)、超迁移实验和非放射性激酶活性检测法检测系膜细胞内AP-1活化、AP-1的组成及JNK活性。应用ELISA法检测培养上清中纤连蛋白(FN)。结果 AngⅡ可诱导MC内JNK活化,刺激30min后,JNK活性达到高峰,1h后几乎恢复至正常水平。AngⅡ刺激后MC内AP-1活性显著增强,活化的AP-1主要含有c-Jun和c-Fos亚基。Ang Ⅱ可促进MC表达TGF-β1及分泌FN,抗TGF-β1抗体显著抑制AngⅡ促进的FN分泌。JNK特异性抑制剂SP600125显著抑制AngⅡ诱导AP-1活化、TGF-β1表达及FN分泌。结论AngⅡ-JNK-c-Jun/AP-1—TGF-β1-FN信号转导通路在肾小球硬化中发挥一定作用,SP600125能部分抑制Ang Ⅱ诱导的AP-1活化、TGF-β1表达及FN的合成,可能具有一定的治疗作用。
Objective To investigate the role of JNK-c-Jun/AP-1 signal pathway in the regulation of Ang Ⅱ-induced transforming growth factor-β1 (TGF-β1) expression in human renal mesangial cells(MC). Methods TGF-β1 mRNA expression was examined by ribonuclease protection assay. Electrophoretic mobility shift assay (EMSA) and non-radioactive kinase assay were used to detect the activity of AP-1 and JNK. Fibronectin was determined by ELISA. Results Ang Ⅱ enhanced the activity of JNK,with a peak at 30 min,and the activity of JNK returned to the basal level after 1 h. EMSA showed that Ang Ⅱ activated AP-1 in MC. Supershift assay demonstrated that c-Jun and c-Fos were the predominant subunits involved. Ang Ⅱ significantly upregulated TGF-β1 mRNA expression and FN excretion. Anti-TGF-β1 antibody inhibited FN excretion in Ang Ⅱ-stimulated MC. Significant down-regulation of AP-1 activation,TGF-β1 mRNA expression and FN excretion were observed in Ang Ⅱ-induced MC pretreated with JNK specific inhibitor SP600125. Conclusion Ang Ⅱ -JNK(c-Jun/AP-1-TGF-β1-FN pathway may play an important role in the glomerulosclerosis.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2003年第5期301-304,共4页
Chinese Journal of Nephrology
基金
国家自然科学基金(30100081)
