SiO_(2)刺激矽肺肺泡巨噬细胞上清液中CTGF在肺泡上皮细胞间充质化的作用  

Effect of CTGF in supernatant of silicosis alveolar macrophages stimulated by SiO_(2) on epithelial-mesenchyme transition of alveolar epithelial cells

作　　者：苏艺伟 唐侍豪 张晋蔚 王建宇[1] 张燕[1] 刘移民[1] SU Yi-wei;TANG Shi-hao;ZHANG Jin-wei;WANG Jian-yu;ZHANG Yan;LIU Yi-min(Occupational Health Evaluation and Testing Center,Guangzhou Tweljih People's Hospital,Guangzhou Guangdong,510620,China)

机构地区：[1]广州市第十二人民医院职业卫生评价检测中心,广东广州510620

出　　处：《职业与健康》2022年第5期599-602,共4页Occupation and Health

基　　金：广州市卫生健康科技项目(20201A011048);广州市高水平临床重点专科建设项目[穗卫函](2019)1555号。

摘　　要：目的了解SiO_(2)体外激活肺泡巨噬细胞上清液结缔组织生长因子(connective tissue growth factor,CTGF)的表达情况及对肺泡上皮细胞间充质化的作用。方法从矽肺患者肺泡灌洗液中分离肺泡巨噬细胞,使用SiO_(2)进行干预,收集处理后6、12、24、48 h上清液,检测各时间点CTGF表达情况;将A549细胞分成3组,分别为空白对照组、未刺激上清液组和刺激上清液组,刺激上清液组加入SiO_(2)干预24 h的肺泡巨噬细胞上清液,未刺激上清液组加入无SiO_(2)干预的24 h肺泡巨噬细胞上清液,空白对照组加入细胞培养基,培养48 h后,检测A549细胞CTGF、E-钙粘蛋白(E-cadherin)和α-平滑肌肌动蛋白(α-SMA蛋白)表达情况。结果刺激组的肺泡巨噬细胞上清液CTGF表达量随SiO_(2)干预时间的延长而升高,于24 h达到峰值[(30.48±1.66)ng/L],除6 h外,其他时间点刺激组上清液CTGF表达量[(21.46±1.77)、(30.48±1.66)、(25.67±2.39)ng/L]均高于同期未刺激组上清液CTGF表达量[(17.36±1.96)、(18.86±1.90)、(18.32±1.64)ng/L],差异均有统计学意义(均P<0.05);在干预48 h后,未刺激上清液组和刺激上清液组的A549细胞呈长条形或不规则形,排列紊乱,刺激上清组细胞变化更明显,甚至出现分叉;刺激上清液组A549细胞CTGF、α-SMA表达量分别高于未刺激组和空白对照组,差异均有统计学意义(均P<0.05/3),而E-cadherin表达量低于未刺激上清液组和空白对照组,差异均有统计学意义(均P<0.05/3)。结论经SiO_(2)激活的肺泡巨噬细胞CTGF的表达量升高,可诱导肺泡上皮细胞增加CTGF和α-SMA的表达、降低E-cadherin的表达,CTGF可能通过旁分泌或自分泌的形式促进肺泡上皮细胞间充质化改变。Objective To investigate the expression of connective tissue growth factor(CTGF)in the supernatant of alveolar macrophage activated by SiO_(2) in vitro and its effect on the epithelial-mesenchyme transition of alveolar epithelial cells.Methods The alveolar macrophages were separated from the alveolar lavage fluid of patients with silicosis and treated by SiO_(2),the supernatant at 6,12,24,and 48 hours after treatment were collected to detect the expression of CTGF at each time point.The A549 cells were divided into three groups,namely the blank control group,the unstimulated supernatant group and the stimulated supernatant group.The stimulated supernatant group was added with alveolar macrophage supernatant treated with SiO_(2) for 24hours,the unstimulated supernatant group was added with alveolar macrophage supernatant without the intervention of SiO_(2) for 24hours,and the blank control group was added with cell culture medium.After 48 hours of culture,the expression of CTGF,E-cadherin andα-SMA protein in A549 cells was detected.Results The expression of CTGF in the supernatant of alveolar macrophages in the stimulated group increased with the extension of the SiO_(2) intervention time,reaching a peak[(30.48±1.66)ng/L]at 24 hours.Except for 6 hours,the expression of CTGF in the supernatant of the stimulated group[(21.46±1.77),(30.48±1.66),(25.67±2.39)ng/L]was higher than that of the unstimulated group at other time points[(17.36±1.96),(18.86±1.90),(18.32±1.64)ng/L],and the differences were statistically significant(P<0.05).After 48 hours of intervention,the A549 cells in the unstimulated supernatant group and the stimulated supernatant group showed long or irregular shapes and arranged disorderly,and the cells in the stimulated supernatant group changed more obviously,and even bifurcated.The expression levels of CTGF andα-SMA in A549 cells in the stimulated supernatant group were higher than those in the unstimulated group and the blank control group,and the differences were statistically significant(a

关 键 词：二氧化硅 矽肺 肺泡巨噬细胞 结缔组织生长因子 上皮间充质化 

分 类 号：R135.2[医药卫生—劳动卫生]