白介素⁃17A通过自噬调控支气管成纤维细胞胶原降解

Interleukin⁃17A regulates collagen degradation in bronchial fibroblasts through autophagy

目的探讨白介素⁃17A(interleukin⁃17A,IL⁃17A)对大鼠支气管成纤维细胞(bronchial fibro⁃blasts,BF)增殖、自噬及自噬后胶原降解的影响。方法采用酶联合消化法+组织块贴壁法培养BF并鉴定,CCK⁃8法检测细胞增殖能力;实验分组:对照组、雷帕霉素组、雷帕霉素+IL⁃17A组、IL⁃17A组;透射电镜观察细胞自噬泡的形成情况;Western blot检测BF中自噬标记蛋白LC3、p62的表达;ELISA法检测各组中基质金属蛋白酶⁃1(matrix metalloproteinase⁃1,MMP⁃1)与基质金属蛋白酶抑制剂(matrix metalloproteinase inhibitor,TIMP⁃1)的表达。结果IL⁃17A浓度为20 ng/mL时BF细胞增殖能力最佳。IL⁃17A组中自噬泡数量明显减少,LC3⁃Ⅱ/LC3⁃I的比值及MMP⁃1的表达水平明显下调,p62及TIMP⁃1的表达水平明显上调(P<0.05)。雷帕霉素部分逆转了IL⁃17A的作用。结论(1)IL⁃17A可促进BF细胞增殖;(2)IL⁃17A通过抑制自噬,发挥抑制胶原降解的作用,促进气道重塑的发生发展。

Objective To investigate the effects of Interleukin⁃17A on rat bronchial fibroblasts prolifera⁃tion,autophagy and collagen degradation after autophagy.Methods Enzyme combined digestion method and tissue block adherence method was used to culture and identify BF.CCK⁃8 method was used to detect cell prolifera⁃tion.Experimental groups:control group,rapamycin group,IL⁃17A+rapamycin group,IL⁃17A group.The forma⁃tion of autophagic vesicles was observed by transmission electron microscopy.The expression of autophagy marker proteins LC3 and p62 in BF were detected by Western blot.ELISA was used to detect matrix metalloproteinase⁃1 and matrix metalloproteinase inhibitor expression.Results The proliferation ability of BF cells was best when the IL⁃17A concentration was 20 ng/ml.The number of autophagic vesicles in the IL⁃17A group was significantly reduced,the ratio of LC3⁃II/LC3⁃I and the expression level of MMP⁃1 were significantly down⁃regulated,and the expression levels of p62 and TIMP⁃1 were significantly up⁃regulated(P<0.05).Rapamycin partially reversed the effects of IL⁃17A.Conclusion IL⁃17A can promote the proliferation of BF cells.IL⁃17A also can inhibit the degradation of collagen by inhibiting autophagy,and promote the occurrence and development of airway remodeling.