JAK-STAT通路参与CTGF诱导人胚肺成纤维细胞转分化的研究

摘要

Objective To investigate the effect of JAK - STAT signal transduction pathway in the transdifferentiation process of re-combinant human CTGF - stimulated human embryonic lung fibroblasts( HFL - I) . Methods The HFL-1 were cultured in vitro and were divided into three equal groups: control group, CTGF group, CTGF and JAK - STAT inhibitor AG490 group. The expression levels of total -STAT3, phosphor - STAT3 and α-SMA in each group were detected by western blot, while RT - PCR was used to test the expression of STAT3 mRNA in each group. Results The levels of p - STAT3 and α- SMA were significantly increased in CTGF group compared with control group( P＜0. 05). And JAK - STAT inhibitor AG490 (50μmol/L) largely down - regulated α-SMA and p - STAT3 expression in response to CTGF(P＜0.05 or P＜0.01). Conclusion JAK - STAT is a upstream mediator in the lung gbroblast - rayofibroblast process stimulated by CTGF,and JAK - STAT inhibitor AG490 can significantly abrogated the effect of CTGF.%目的 研究JAK - STAT信号通路在重组人结缔组织生长因子(recombinant human,CTGF)诱导人胚肺成纤维细胞(HFL -I)转分化过程中的作用.方法 将体外培养的HFL -I分为正常对照组、CTGF组及CTGF +JAK -STAT特异性抑制剂通路抑制剂AG490组,选取不同时相点,Western blotting测定平滑肌肌动蛋白(α-smooth muscle actin,α- SMA)、磷酸化蛋白(p- STAT3)和总蛋白STAT3的蛋白表达,RT - PCR检测STAT3 mRNA表达.结果 正常体外培养的HFL -I有微量的p -STAT3、α- SMA表达,加入20ng/ml CTGF诱导15min后p- STAT3水平明显升高(P＜0.05),30min时升至顶峰后逐渐减弱；诱导24h后α- SMA表达显著升高(P＜0.05).AG490 50μmol/L预处理60min,p - STAT3和α- SMA表达明显抑制(P＜0.05或P＜0.01).结论 CTGF诱导HFL -I转分化为肌成纤维细胞,JAK - STAT通路参与上述过程,其特异性抑制剂AG490可有效抑制该效应.