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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。
Objective To investigate the effect of TGF-β1 on the expression of heat shock protein 47 (HSP47) and type Ⅰ collagen (Collagen Ⅰ) in human embryonic lung fibroblasts (HELF) and to explore the role of TGF-β1 in the formation of pulmonary fibrosis. Methods TGF-β1 was used to stimulate HELF to observe the dynamic expression of HSP47 and collagenⅠ. Group A was treated with TGF-β15ng / mL at the same concentration for 10h, 12h, 24h, 48h and 72h, β10ng / ml, 1ng / ml, 5ng / ml, 10ng / ml for 48h. Western blotting was used to detect the expression of HSP47 and collagenⅠ in group a and group b. Morphological changes and proliferation were observed by inverted phase contrast microscope. Results Western blot analysis showed that the expression of HSP47 and Collagen Ⅰ increased synchronously with the increase of time under the action of 5ng / mL TGF-β1 in HELF group (P <0.05), while the expression of HSP47 and collagenⅠ in group b increased gradually Also increased simultaneously (P <0.05). The inverted phase contrast microscope showed that HELF cells became spindle-shaped growth, and proliferated significantly after addition of TGF-β1. Conclusion TGF-β1 may regulate the secretion and secretion of Collagen Ⅰ mainly by promoting the secretion of HSP47. HSP47 plays a crucial role in the development of pulmonary fibrosis as a collagen-specific chaperone and may be a new therapeutic target for pulmonary fibrosis.