线粒体分裂、线粒体自噬在烟草提取物诱导的支气管上皮细胞损伤模型中的作用  

The role of mitochondrial fission and mitophagy in cigarette smoke extract-induced lung epithelial cell injury models

作　　者：王素文 宋小敏 李忱菲 魏良煜 张海[2] 李锋 王炯[1] Wang Suwen;Song Xiaomin;Li Chenfei(Dept of Geriatric Respiratory and Critical Care Medicine,The First Affiliated Hospital of Anhui Medical University,Hefei 230022;Dept of Pulmonary and Critical Care Medicine,Shanghai Chest Hospital,Shanghai Jiao Tong University,Shanghai 200030)

机构地区：[1]安徽医科大学第一附属医院老年呼吸与危重症医学科,合肥230022 [2]上海市胸科医院(上海交通大学附属胸科医院)呼吸与危重医学科,上海200030

出　　处：《安徽医科大学学报》2021年第9期1343-1349,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82070041)。

摘　　要：目的探讨线粒体分裂、线粒体自噬在烟草提取物(CSE)诱导的支气管上皮细胞损伤模型中的作用。方法支气管上皮细胞与5%CSE共培养,使用线粒体分裂抑制剂Mdivi-1(Md)和线粒体自噬促进剂Urolithin A(UA)进行预处理,分别检测细胞增殖活力、氧化应激、线粒体结构变化、炎症因子(IL-1β、IL-6、IL-18、CXCL1、CXCL8)mRNA水平、细胞程序性坏死组分(RIPK1、RIPK3、MLKL)mRNA水平、线粒体分裂蛋白(DRP1、MFF)和线粒体自噬蛋白(p62/SQSTM1、LC3B)的水平。结果5%CSE诱导氧化应激、炎症因子mRNA增加、线粒体网络结构破坏、线粒体分裂蛋白表达增加、线粒体自噬蛋白p62/SQSTM1蛋白表达增高、LC3BⅡ/Ⅰ蛋白表达下降、程序性坏死组分mRNA增加。Md/UA预处理可抑制氧化应激,抑制炎症因子、细胞程序性坏死组分mRNA表达,部分恢复线粒体网络结构,降低线粒体分裂蛋白水平。Md预处理增加线粒体自噬蛋白p62/SQSTM1蛋白水平,不影响LC3BⅡ/Ⅰ蛋白表达。UA预处理抑制线粒体自噬蛋白p62/SQSTM1蛋白水平,增加LC3BⅡ/Ⅰ蛋白表达。结论抑制线粒体分裂或促进线粒体自噬可抑制烟草暴露诱导的氧化应激和炎症反应,部分恢复线粒体结构,其机制可能与抑制线粒体分裂、抑制细胞程序性坏死组分mRNA表达有关。Objective To investigate the role of mitochondrial fission and mitophagy in the model of bronchial epithelial cell injury induced by cigarette smoke extract.Methods Bronchial epithelial cells were co-cultured with 5%CSE,pretreated with mitochondrial fission inhibitor Mdivi-1(Md)and mitophagy inducer Urolithin A(UA).Cell proliferation,oxidative stress,mitochondrial structure,the mRNA levels of inflammatory factor(IL-1,IL-6,IL-18,CXCL 1,CXCL 8)and necroptosis components(RIPK1,RIPK3,MLKL),mitochondrial fission protein(DRP1,MFF)and mitophagy protein(p62/SQSTM1,LC3B)were examined.Results In Beas 2b cells,5%CSE induced oxidative stress,increased the mRNA expression of inflammatory factor and necroptosis components,induced the destruction in mitochondrial network,increased the protein expression of mitochondrial fission and mitophagy protein p62/SQSTM1 expression and decreased mitophagy protein LC3BⅡ/Ⅰexpression.Md/UA pretreatment inhibited oxidative stress,inhibited the expression of inflammatory factor mRNA and necroptosis components mRNA,partially restored mitochondrial network structure and decreased mitochondrial fission protein level.Md pretreatment increased mitophagy protein p62/SQSTM1 protein level,but did not affect LC3BⅡ/Ⅰprotein expression.Pretreatment with UA inhibited p62/SQSTM1 protein expression,and increased LC3BⅡ/Ⅰprotein expression.Conclusion Inhibiting mitochondrial fission or promoting mitophagy can reduce CSE-induced oxidative stress and inflammatory response,and restore mitochondrial structure.The mechanism may be related to the inhibition of mitochondrion fission and the mRNA expression of necroptosis components.

关 键 词：线粒体分裂 线粒体自噬 烟草暴露 炎症 程序性坏死 

分 类 号：R563.3[医药卫生—呼吸系统]