二氧化硅通过调节人气道上皮细胞自噬促进MUC5AC表达  

SiO_2 increases MUC5AC expression by regulating autophagy in human airway epithelia cells

作　　者：王孝芸 唐红梅 王星 马宁 李月蛟 袁谢芳 徐国锋 张沄 WANG Xiaoyun;TANG Hongmei;WANG Xing;MA Ning;LI Yuejiao;YUAN Xiefang;XU Guofeng;ZHANG Yun(Inflammation&Allergic Diseases Research Unit,Affiliated Hospital of Southwest Medical University,Luzhou 646000,China;不详)

机构地区：[1]西南医科大学附属医院炎症与变态反应实验室,四川泸州646000 [2]西南医科大学附属医院呼吸与危重症医学科,四川泸州646000

出　　处：《实用医学杂志》2023年第3期303-308,共6页The Journal of Practical Medicine

基　　金：国家自然科学基金项目(编号:81900028)。

摘　　要：目的 探讨二氧化硅(SiO_(2))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℃和5%CO_(2)的培养箱培养Beas-2b。使用NC-siRNA、ATG5-siRNA、BECN1-siRNA及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(1μg/mL)进行筛选,获得稳定转染细胞株。用SiO_(2)(100μg/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 SiO_(2)组的MUC5AC、ATG5、BECN1和LC3Ⅱ的表达高于对照组(P <0.05);ATG5和BECN1敲减细胞株中,SiO_(2)刺激后MUC5AC和LC3Ⅱ的表达均低于对照组(P <0.05)。结论 SiO_(2)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和BECN1信号途径相关。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(1μg/mL)was used to obtain stable transfected cell line.Cells were treated with SiO2(100μg/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 LC3Ⅱin cells treated by SiO2 was signifiacntly higher than that in control cells(P<0.05).After stimulation of SiO2,expression of MUC5AC and LC3Ⅱin 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.

关 键 词：二氧化硅 自噬 黏蛋白5AC ATG5 BECN1 

分 类 号：R563[医药卫生—呼吸系统]