热休克蛋白47参与TGF-β1促人胚肺细胞胶原合成

摘要

目的 观察热休克蛋白47(HSP47)对人胚肺成纤维细胞(HELF)胶原合成及间质细胞标志物的影响,探讨它与肺纤维化的关系.方法 用TGF-β1诱导HELF表达HSP47,用5 ng/mL TGF-β1作用不同时间和用不同浓度TGF-β1作用于HELF 48 h;WST-8法检测细胞增殖;免疫荧光法检测HSP47的表达;Western blot检测HsP47、Ⅰ型胶原(Collage 1)及间质细胞标志物α-平滑肌肌动蛋白(α-SMA)、波形蛋白(Vimentin)的表达.结果 正常HELF表达少量HSP47,随着HELF在TGF-β1作用时间的延长和剂量逐渐增加,HSP47(5 ng/mL TGF-β1作用48 h,7.081±0.265)、Collage Ⅰ(5 ng/mL TGF-β1作用48 h,2.494±0.139)、α-SMA和Vimentin的表达均同步增强(P<0.05).结论 HSP47在TGF-β1的作用下,促进人胚肺成纤维细胞胶原蛋白的合成,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.%Objective By investigating effect of the dynamic expression of HSP47 in human embryonic lung fibroblast (HELF) on pulmonary fibrosis to explore the relation between HSP47 and pulmonary fibrosis. Methods Dynamic expression of HSP47, CollageI, a-SMA and Vimentin were observed after HSP47 was induced to express in HELF with incubation of TGF-β1. HELF was stimulated by TGF-β1 (5 ng/mL) at different time points and different dosages for 48 h respectively. The proliferative rate of HELF was tested by WST. Expression of HSP47 was determined by immunofluorescence. Expressions of HSP47, CollageI, a-SMA and Vimentin were detected by Western blot. Results HSP47 showed slight expression in normal HELF. Expressions of HSP47(5 ng/mL TGF-β1 48 h,7. 081 ±0. 265),CollageI(5 ng/mL TGF-β1 48 h,2. 494 ±0. 139) ,a-SMA and Vimentin increased synchronously by means of time-in-dependence and dosage-independence after HELF being treated by TGF-β1 (P ＜0. 05). Conclusion HSP47 participated in the expression of collagen in human embryonic lung fibroblast induced by TGF-β1, suggested that HSP47 might play an important role in pulmonary fibrosis as collagen-specific molecular chaperone.