转位蛋白配体XBD173对香烟烟雾提取物诱导小鼠肺炎症反应的减轻作用及其机制  

Attenuating effect of translocator protein ligand XBD173 on pulmonary inflammatory response induced by cigarette smoke extraction in mice and its mechanism

作　　者：高兴洪 唐红梅 李月蛟 王孝芸 王星 袁谢芳 吴敏 GAO Xinghong;TANG Hongmei;LI Yuejiao;WANG Xiaoyun;WANG Xing;YUAN Xiefang;WU Min(Laboratory of Inflammation and Allergy,Affiliated Hospital,Southwest Medical University,Luzhou 646000,China)

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

出　　处：《吉林大学学报（医学版）》2023年第2期272-279,共8页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金青年基金项目(82001703)。

摘　　要：目的:探讨转位蛋白(TSPO)配体XBD173对香烟烟雾提取物(CSE)诱导小鼠肺炎症反应和巨噬细胞M1型极化的影响,阐明其相关分子机制。方法:18只成年雄性C57BL/6小鼠随机分为对照组、CSE组和CSE+XBD173组,CSE组小鼠采取20μL CSE滴鼻,CSE+XBD173组小鼠给予相同剂量CSE和腹腔注射10 mg·kg^(-1)XBD173。采用HE染色和PAS染色观察各组小鼠肺组织病理形态表现。培养小鼠RAW264.7巨噬细胞,分为对照组、CSE组和CSE+XBD173组,CSE给药浓度为1%,XBD173给药浓度为4 mg·L^(-1),对照组给予相同体积的二甲基亚砜;刺激18 h后,流式细胞术检测各组RAW264.7细胞中CD80、CD86、诱导型一氧化氮合成酶(iNOS)和活性氧(ROS)表达水平及细胞凋亡率,Western blotting法检测各组RAW264.7细胞中核因子κB/p65(NF-κB/p65)和磷酸化NF-κB/p65(p-NF-κB/p65)蛋白表达水平。采用siRNA敲低RAW264.7细胞中TSPO表达,实时荧光定量PCR(RT-qPCR)法检测各组细胞中白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)mRNA表达水平。结果:动物实验,与对照组比较,CSE组小鼠肺组织中炎症细胞数量明显增多、支气管管壁增厚;与CSE组比较,XBD173+CSE组小鼠肺组织中炎症细胞减少,支气管管壁变薄。细胞实验,与对照组比较,CSE组RAW264.7细胞中CD80、CD86、iNOS和ROS表达水平明显升高(P<0.05),细胞凋亡率明显升高(P<0.05),IL-6和TNF-αmRNA表达水平升高(P<0.05),p-NF-κB/p65蛋白表达水平明显升高(P<0.05),NF-κB/p65蛋白表达水平差异无统计学意义(P>0.05);与CSE组比较,XBD173+CSE组RAW264.7细胞中CD80、CD86、iNOS和ROS表达水平明显降低(P<0.05),细胞凋亡率明显降低(P<0.05),IL-6和TNF-αmRNA表达水平明显降低(P<0.05),p-NF-κB/p65蛋白表达水平明显降低(P<0.05)。siRNA敲低RAW264.7细胞TSPO蛋白表达后,各组细胞中IL-6和TNF-αmRNA表达水平比较差异无统计学意义(P>0.05)。结论:XBD173可减轻CSE引起的小鼠肺炎症反应,抑制CSE诱导的巨噬细胞M1极化Objective:To investigate the effects of translocator protein(TSPO)ligand XBD173 on the pulmonary inflammatory response and M1 polarization of macrophages of the mice induced by cigarette smoke extraction(CSE),and to clarify their realted molecular mechanisms.Methods:A total of 18 adult male C57BL/6 mice were randomly divided into control group,CSE group,and CSE+XBD173 group.The mice in CSE group were intranasally given 20μL CSE,and the mice in CSE+XBD173 group were given the same dose of CSE and intraperitoneally injectied with 10 mg·kg^(-1) XBD173.The pathomorphology of lung tissue of the mice in various groups was observed by HE staining and PAS staining.The RAW264.7 macrophage were cultured and divided into control group,CSE group,and CSE+XBD173 group.The CSE administration concentration was 1%,the XBD173 administration concentration was 4 mg·L^(-1),and the cells in control group were given the same volume of dimethyl sulfoxide;after 18 h of stimulation,the expression levels of CD80,CD86,inducible nitric oxide synthase(iNOS),reactive oxygen species(ROS)in the RAW264.7 cells,and the apoptotic rates of the RAW264.7 cells in various groups were detected by flow cytometry;Western blotting method was used to detect the expression levels of nuclear factor-κB/p65(NF-κB/p65)and phosphorylated NF-κB/p65(p-NF-κB/p65)proteins in the cells in various groups.The siRNA was used to knock down the TSPO expression in the RAW264.7 cells in various groups,and the expression levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)mRNA in the cells in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)method.Results:The animal experiment results showed that compared with control group,the number of inflammatory cells in lung tissue of the mice in CSE group was increased significantly and the bronchial wall was thicker;compared with CSE group,the number of inflammatory cells in lung tissue of the mice in XBD173+CSE group was decreased and the bronchial wall was thinner.The cell experimen

关 键 词：转位蛋白 XBD173 烟草提取物 巨噬细胞 核因子ΚB 

分 类 号：R392.12[医药卫生—免疫学]