TGF-β1对人胚肺成纤维细胞HSP47、collagenⅠ表达的影响

Effects of TGF-β1 on the expression of HSP47 and collagenⅠin HELF

目的通过观察TGF-β1对人胚肺成纤维细胞(HELF)的热休克蛋白47(HSP47)及Ⅰ型胶原(CollagenⅠ)表达的影响,探讨三者在肺纤维化形成中的作用和关系。方法用TGF-β1刺激HELF,观察HSP47、collagenⅠ的动态表达;实验分2组:a组相同浓度TGF-β15ng/mL作用于细胞后0h,12h,24h,48h,72h;b组不同浓度TGF-β10ng/ml、1ng/ml、5ng/ml、10ng/ml作用于细胞48h;免疫印迹技术检测a组与b组的HSP47、collagenⅠ的表达情况。通过倒置相差显微镜观察细胞形态学变化及增殖情况。结果免疫印迹分析:a组HELF在5ng/mLTGF-β1作用下,HSP47与CollagenⅠ的表达随着时间的延长同步增强(P<0.05);b组TGF-β1剂量逐渐增加后,HSP47与collagenⅠ的表达亦同步增强(P<0.05)。倒置相差显微镜下可见HELF细胞成梭形生长,加入TGF-β1后明显增殖。结论 TGF-β1可能主要通过促进HSP47分泌调控CollagenⅠ的合成分泌,HSP47作为胶原特异性分子伴侣在肺纤维化的发生发展过程中发挥至关重要的作用,有望成为治疗肺纤维化的新靶点。

Aim To investigate the effect of TGF-β1 on the expression of heat shock protein 47（HSP47）and collagen Ⅰin human embryonic lung fibroblast（HELF）. Methods Dynamic expression of HSP47 and collagenⅠwere observed after heat shock protein 47 was induced to express in HELF in incubation of TGF-β1. Experiment objects were divided into two groups （group a and group b）. Group a was stimulated by TGF-β1 at different time points（0h,12h,24h,48h,72h）,Group b in different dosages （0ng/ml,1ng/ml,5ng/ml,10ng/ml）.Expressions of HSP47 and collagen Ⅰ were detected by Western blot. Results Group a showed expressions of HSP47 and collagenⅠincreased synchronously in Western blot by means of time-independence after HELF being treated by TGF-β1 （P〈0.05）; as well as Group b by means of dosage-independence （P〈0.05）. HELF showed a fusiform growth under inverted phase contrast microscope and proliferation in incubation of TGF-β1. Conclusions TGF-β1 might regulate synthesis of collagenⅠmainly by promoting secretion of HSP47,that suggested HSP47 might play an important role in pulmonary fibrosis as collagen-specific molecular chaperone.It was expected to be a new target for treatmetn of pulmonary fibrosis.