下调HMGB1表达通过抑制炎性反应与上皮间质转化改善慢性阻塞性肺疾病小鼠疾病进展  

Down-regulation of HMGB1 expression ameliorates the progression of COPD in mice by inhibiting inflammatory response and epithelial mesenchymal transition

作　　者：热依拉·牙合甫[1] 买热木古·阿不都热依木 王琴[1] 李瑞晟[1] 穆清爽[1] REYILA·Yahefu;MAIREMUGU·Abudureyimu;WANG Qin;LI Ruisheng;MU Qingshuang(The Seoond Afiliated Hospital of Xinjiang Medical University Geratrics(cadre ward),Unumqi 830063,China)

机构地区：[1]新疆医科大学第二附属医院老年病(干部病房),新疆乌鲁木齐830063

出　　处：《细胞与分子免疫学杂志》2022年第8期685-691,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：新疆维吾尔自治区自然科学基金(2019D01C235)。

摘　　要：目的在慢性阻塞性肺疾病(COPD)的小鼠模型中探究下调高迁移率族蛋白B1(HMGB1)表达对肺组织中炎性反应及上皮间质转化(EMT)的影响和相关机制。方法采用单纯香烟烟雾诱导建立小鼠COPD模型,使用小干扰RNA(siRNA)下调小鼠肺组织中HMGB1表达,并按实验目的将小鼠分为对照组、COPD组、si-NC处理组(COPD联合si-NC组),si-HMGB1处理组(COPD联合si-HMGB1组),并设立噻托溴铵(tiotropium)处理组(COPD联合tiotropium组)作为阳性对照组。香烟烟雾诱导4周后,观察小鼠的一般情况差异,并记录每周各组小鼠的体质量变化;HE染色观察各组小鼠肺组织病理改变;实时荧光定量PCR和Western blot法分别检测小鼠肺组织HMGB1 mRNA和蛋白表达;采集各组小鼠支气管肺泡灌洗液(BALF),ELISA检测BALF中白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)、IL-1β、转化生长因子β1(TGF-β1)的表达水平;免疫组织化学染色观察各组小鼠肺组织上皮钙黏蛋白(E-cadherin)和α平滑肌肌动蛋白(α-SMA)的表达;Western blot法检测肺组织E-cadherin、α-SMA和HMGB1/核因子κB(NF-κB)信号通路中晚期糖基化终产物受体(RAGE)、Toll样受体4(TLR4)、TGF-β1表达和NF-κB p65磷酸化水平。结果成功建立香烟烟雾诱导的COPD小鼠模型,与COPD组相比,下调肺组织中HMGB1表达能显著改善小鼠的一般生命体征,促进其体质量增加,并能够改善小鼠肺组织病理学损伤;与对照组相比,COPD组、COPD联合si-NC组和COPD联合tiotropium组小鼠肺组织中HMGB1 mRNA和蛋白表达水平均明显增加,而COPD联合si-HMGB1组无明显HMGB1表达;且与对照组相比,COPD造模组小鼠BALF中IL-6、TNF-α、IL-1β、TGF-β1的分泌水平、肺组织中α-SMA、RAGE、TLR4、TGF-β1表达和NF-κB p65磷酸化水平均显著增加,而E-cadherin表达水平明显降低;而与COPD组或COPD联合si-NC组相比,COPD联合si-HMGB1组和COPD联合tiotropium组小鼠肺组织中上述指标变化均显著降低,后两Objective To explore the effect of down-regulation of high mobility group box 1(HMGB1)expression on inflammatory response and epithelial mesenchymal transition(EMT)in the lung tissue of mice with chronic obstructive pulmonary disease(COPD)and its related mechanism.Methods The COPD model was induced by cigarette smoking,and HMGB1 expression in lung tissue of mice was down-regulated by small interfering RNA(siRNA).The mice were divided into negative control group(NC group),COPD group,si-NC intervention COPD group,si-HMGB1 intervention COPD group,and tiotropium bromide intervention COPD group(positive control group).After 4 weeks of cigarette smoking induction,the general condition of mice were observed,and the body mass changes of each group were recorded every week.HE staining was used to observe the pathological changes of lung tissue in each group.The expression of HMGB1 mRNA and protein in lung tissues of mice weredetected by real time quantitative PCR and Western blot analysis.The BALF of mice in each group wascollected,and the levels of IL-6,TNF-α,IL-1βand TGF-β1 in BALF were detected by ELISA.The expressions of E-cadherin andα-SMA in lung tissues of mice were observed by immunohistochemical staining.The expression of RAGE,TLR4,TGF-β1 and the phosphorylation of NF-κB p65 in lung tissues were detected by Western blot analysis.Results COPD mouse model induced by cigarette smoking was successfully established.Compared with COPD group,down-regulation of HMGB1 expression in lung tissue significantly improved the general vital signs of mice,promoted the increase of body mass,and improved the pathological damage of lung tissue in mice.Compared with the control group,HMGB1 mRNA and protein expression levels increased significantly in COPD group,COPD combined with si-NC group and COPD combined with tiotropium group,while no significant HMGB1 expression was detected in COPD combined with si-HMGB1 group.Compared with the control group,the secretion levels of IL-6,TNF-α,IL-1β,TGF-β1 in BALF,the expression levels

关 键 词：慢性阻塞性肺疾病(COPD) 高迁移率族蛋白B1(HMGB1) 上皮间质转化(EMT) 香烟烟雾 

分 类 号：R563[医药卫生—呼吸系统] R364.5[医药卫生—内科学] R392-33[医药卫生—临床医学]