二氧化硅致肺泡巨噬细胞自噬对其自身分泌肿瘤坏死因子-α和转化生长因子-β的影响  

Effects of SiO2-induced autophagy in alveolar macrophage on secretion of TNF-α and TGF-β

作　　者：朱会兴 高俊玲[1,2,3] 赵曼曼[1] 李冉[1] 田艳霞[1] 王鑫[1] 张娟[1] 崔建忠[4] 

机构地区：[1]河北联合大学基础医学院组织学与胚胎学教研室,唐山063000 [2]唐山市重点实验室,唐山063000 [3]河北省煤矿卫生与安全实验室,唐山063000 [4]唐山市工人医院神经外科,唐山063000

出　　处：《解剖学杂志》2015年第1期8-11,共4页Chinese Journal of Anatomy

基　　金：河北省科技支撑重点专项(09276191D);河北省重点职业病防治技术研究(13277709D)

摘　　要：目的：研究自噬抑制剂3-甲基腺嘌呤（3-MA）对二氧化硅（SiO2）诱导的肺泡巨噬细胞自噬的激活,是否对自身分泌肿瘤坏死因子-α（TNF-α）和转化生长因子-β（TGF-β）产生影响.方法：收集SD雄性大鼠肺泡巨噬细胞灌洗液,将其随机分为对照组,SiO2刺激组（SiO2组）,自噬抑制剂组（3-MA组）.采用体外暴露肺泡巨噬细胞染尘法,染尘成功后12 h给予10 mmol/L 3-MA干预.分别于干预后6、18、24h和36 h用胰酶消化并收集体外培养的肺泡巨噬细胞.采用H-E染色观察肺泡巨噬细胞形态学改变;免疫细胞化学检测肺泡巨噬细胞中TNF-α和TGF-β的定位表达;免疫印迹检测肺泡巨噬细胞中TNF-α和TGF-β的蛋白定量.结果：SiO2组与对照组相比肺泡巨噬细胞体积增大,胞质丰富,部分细胞胞质内可见矽尘吞噬颗粒.3-MA组较SiO2组肺泡巨噬细胞形态改变减轻.与对照组相比,SiO2组各时间相肺泡巨噬细胞中TNF-α和TGF-β的蛋白表达显著升高,于SiO2刺激后6h开始升高,18h达高峰,24 h与36 h回落,但仍高于对照组水平.3-MA组可以显著下调SiO2组对应时间点肺泡巨噬细胞中TNF-α和TGF-β的表达.结论：自噬抑制剂3-MA抑制肺泡巨噬细胞自噬后可以下调肺泡巨噬细胞中TNF-α和TGF-β的表达及释放,进而促进炎症损伤的修复。Objective： To observe effects of SiO2-induced alveolar macrophage （AM） autophagy on the expression of TNF-a and TGF-β. Methods： AM obtained from bronchoalveolar lavage was randomly divided into three groups： control group, SiO2 stimulation group, and autophagy inhibitor group （3-MA group）. AM was exposed sliea to stain dust in vitro, and after 12 h in vitro transfection dust, AM was given autophagy inhibitor 3-methyladenine（3-MA） intervention （10 retool/L）. AM was digested and collected in vitro at 6 h, 18 h, 24 h and 36 h. The morphological changes of AM were observed by HE staining. The expressions of TNF-a and TGF-β in AM were detected by immunocytochemistry and Western blotting. Results： Compared to the Control group, the volume of AM in SiO2 group increased, abundant cytoplasm, visible part of the cell cytoplasm engulfed particles of silica dust. Compared to the SiO2 group, the morphological changes of AM in 3-MA group were reduced. Compared to the control group, the expressions of TNF-a and TGF-~ in SiO2 group were significantly increased, both beginning to increase at 6 h, reaching the peak at 18 h, and reducing at 24 h and 36 h, but still higher than base level. The expressions of TNF-a and TGF-~ in 3-MA group were significantly reduced compared to SiO2 group. Conclusion： 3-MA can be reduced in the expression of TNF-a and TGF-~ in AM, and the inhibition of autophagy contributed to inflammatory injury repair.

关 键 词：二氧化硅 肺泡巨噬细胞 自噬 3-甲基腺嘌呤 肿瘤坏死因子-A 转化生长因子-Β 

分 类 号：R563[医药卫生—呼吸系统]