实用医学杂志 ›› 2021, Vol. 37 ›› Issue (4): 423-427.doi: 10.3969/j.issn.1006⁃5725.2021.04.002

• 基础研究 • 上一篇    下一篇

白介素⁃17A通过自噬调控支气管成纤维细胞胶原降解

姚培学,杨胜红, 陈杰, 欧阳瑶   

  1. 遵义医科大学附属医院PCCM 呼吸一病区(贵州遵义 563000)
  • 出版日期:2021-02-25 发布日期:2021-02-25
  • 通讯作者: 欧阳瑶 E⁃mail:ouyangyao116@sohu.com
  • 基金资助:

    国家自然科学基金资助项目(编号:82060005);贵 州省科技计划项目(编号:黔科合支撑 S﹝2020﹞2378);贵州省研 究生科研基金资助项目(编号:黔教合 YJSCXJH﹝2019﹞092 );遵 义市科技支撑计划项目(编号:遵市科合社字(2018)71)

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

YAO Peixue YANG Shenghong,CHEN Jie,OUYANG Yao.   

  1. Department of PCCM Respiratory First Ward,Affiliated Hospital of ZunYi Medical University,Zunyi 563000,China

  • Online:2021-02-25 Published:2021-02-25
  • Contact: OUYANG Yao E⁃mail:ouyangyao116@sohu.com

摘要:

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 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 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

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Abstract:

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

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