实用医学杂志 ›› 2023, Vol. 39 ›› Issue (3): 303-308.doi: 10.3969/j.issn.1006⁃5725.2023.03.007

• 基础研究 • 上一篇    下一篇

二氧化硅通过调节人气道上皮细胞自噬促进 MUC5AC 表达 

王孝芸1 唐红梅1 王星1 马宁1 李月蛟1 袁谢芳1 徐国锋1 张沄1,2    

  1. 西南医科大学附属医院1 炎症与变态反应实验室,2 呼吸与危重症医学科(四川泸州 646000)

  • 出版日期:2023-02-10 发布日期:2023-02-10
  • 通讯作者: 张沄 E⁃mail:zhangyun000hf@163.com
  • 基金资助:
    国家自然科学基金项目(编号:81900028)

SiO2 increases MUC5AC expression by regulating autophagy in human airway epithelia cells

WANG Xiaoyun*,TANG Hongmei,WANG Xing,MA Ning,LI Yuejiao,YUAN Xiefang,XU Guofeng,ZHANG Yun.   

  1. Inflammation & Allergic Diseases Research Unit,Affiliated Hospital of Southwest Medical University,Luzhou 646000,China 

  • Online:2023-02-10 Published:2023-02-10
  • Contact: ZHANG Yun E⁃mail:zhangyun000hf@163.com

摘要:

目的 探讨二氧化硅(SiO2)对人气道上皮细胞株(Beas⁃2b)黏蛋白5AC(MUC5AC)表达的影响 及其作用机制。方法 用含 10%胎牛血清的 DMEM 高糖培养基在 37 ℃ 5% CO2的培养箱培养 Beas⁃2b 使用 NC⁃siRNA、ATG5⁃siRNA、BECN1⁃siRNA mCherry⁃EGFP⁃LC3 等慢病毒转染 Beas⁃2b。各转染组细胞 用相应慢病毒(MOI = 20)转染 12 h 后换新鲜培养基,并用嘌呤霉素(1 μg/mL)进行筛选,获得稳定转 染细胞株。用 SiO2 (100 μg/mL)刺激前述稳定转染细胞株 24 h,并设 PBS 对照组。免疫荧光和免疫印 迹检测 MUC5AC、自噬相关蛋白 5(ATG5)、BECN1 和微管相关蛋白轻链 3(LC3)的蛋白水平;共聚焦显 微镜观察人气道上皮细胞自噬情况。结果 SiO2组的 MUC5AC、ATG5、BECN1 LC3 II 的表达高于对照 组(P < 0.05);ATG5 BECN1 敲减细胞株中,SiO2 刺激后 MUC5AC LC3 II 的表达均低于对照组(P < 0.05)。结论 SiO2可通过调节人气道上皮细胞自噬促进 MUC5AC 表达,其作用机制与 ATG5 BECN1 信号途径相关。

关键词:

二氧化硅, 自噬, 黏蛋白5AC, ATG5, BECN1

Abstract:

Objective To investigate the effect of SiO2 on expression of MUC5AC in Beas⁃2b and explore the related mechanism. Methods Beas⁃2b was cultured in DMEM containing 10% fetal bovine serum at 37 ℃ and 5% CO2. Beas⁃2b was transfected with lentiviruses such as NC⁃siRNA,ATG5⁃siRNA,BECN1⁃siRNA and mcherry⁃ EGFP⁃LC3. The cells in each group were transfected with corresponding lentivirus(MOI = 20)for 12 hours,then with fresh culture medium. Puromycin(1 μg/mL)was used to obtain stable transfected cell line. Cells were treated with SiO2(100 μg/mL)and PBS for 24 hours. Then protein of MUC5AC,ATG5,BECN1 and LC3 was detected by immunofluorenscent test and western blot. Results Expression of MUC5AC,ATG5,BECN1 and LC3Ⅱ in cells treated by SiO2 was signifiacntly higher than that in control cells(P < 0.05). After stimulation of SiO2 expression of MUC5AC and LC3Ⅱ in cells transfected with ATG5⁃siRNA and BECN1⁃siRNA was lower than that in control cells(P < 0.05). Conclusion SiO2 can enhance the expression of MUC5AC in human airway epithelial cells by promoting autophagy,which is related with the regulation of ATG5 and BECN1.

Key words: SiO2,  , autophagy,  , MUC5AC,  , ATG5,  , BECN1