结缔组织生长因子对石英粉尘诱导人支气管上皮细胞释放IL-17A的影响  

Effect of connective tissue growth factor on IL-17A expression in human bronchial epithelial cells induced by crystalline silica dust

作　　者：常薇 雷璐 王震宇[2] 白云斌 冉钶 石玉琴 周婷 刘跃伟[3] CHANG Wei;LEI Lu;WANG Zhen-yu;BAI Yun-bin;RAN Ke;SHI Yu-qin;ZHOU Ting;LIU Yue-wei(School of Public Health, Medical College, Wuhan University of Science and Technology, Wuhan, Hubei 430065, China)

机构地区：[1]武汉科技大学医学院公共卫生学院,湖北武汉430065 [2]武汉科技大学医学院基础医学院 [3]湖北省疾病预防控制中心应用毒理湖北省重点实验室

出　　处：《环境与健康杂志》2018年第5期398-401,共4页Journal of Environment and Health

基　　金：国家自然科学基金(81402659);湖北省自然科学基金(2014CFB811;2016CFB520)

摘　　要：目的探讨结缔组织生长因子(CTGF)对石英粉尘诱导人支气管上皮细胞(16HBE)表达白介素17A(IL-17A)水平的影响。方法采用0、12.5、25、50、100μg/ml石英粉尘染毒人支气管上皮细胞(16HBE)24、48 h,采用CCK-8法测定细胞存活率,以荧光定量PCR方法检测细胞内CTGF mRNA相对表达水平;用CTGF siRNA干扰16HBE细胞12 h后,加入0、25、50μg/ml石英粉尘染毒48 h,并设同样剂量石英粉尘染毒的16HBE细胞为对照组,收集细胞上清液,采用酶联免疫吸附法(ELISA)检测细胞上清液中IL-17A的含量。结果与24 h染尘组比较,石英粉尘染毒16HBE细胞48 h后细胞毒性作用更明显,其细胞存活率随石英粉尘浓度的增加逐渐降低(P<0.05);同时细胞内CTGF mRNA相对表达水平和细胞分泌IL-17A的水平均显著升高(P<0.05),且呈剂量-效应关系,50μg/ml染毒组细胞内表达CTGF mRNA和分泌IL-17A的水平达到最高;与同样剂量石英粉尘染毒的16HBE细胞相比,25、50μg/ml石英粉尘诱导CTGF siRNA干扰组细胞分泌IL-17A的水平显著降低,差异具有统计学意义(P<0.05)。结论抑制CTGF可显著降低石英粉尘诱导16HBE细胞表达IL-17A的水平,提示CTGF可能参与调节石英粉尘致肺部炎性及纤维化的反应过程。Objective To investigate the effects of connective tissue growth factor（CTGF） on interleukin-17 A（IL-17 A）expression in human bronchial epithelial cells（16 HBE） induced by silica dust. Methods 16 HBE cells were exposed to 0, 12.5,25, 50 and 100 μg/ml silica dust for 24 and 48 h, the cell viability was detected by CCK-8 kit,the relative expression level of CTGF mRNA in cells was determined by q RT-PCR. Then 16 HBE cells interfered with CTGF siRNA for 12 h were treated with 0,25 and 50 μg/ml silica dust for 48 h, the levels of IL-17 A were determined by ELISA kits. Results The toxic effects of silica dust on 16 HBE cells was more severe at 48 h time point than that at 24 h time point, showing that the cell viability was significantly decreased induced by silica dust in a concentration-dependent manner（P〈0.05）. The levels of CTGF mRNA expression and IL-17 A secretion in silica-exposed 16 HBE cells were significantly increased in a dose-response manner（P〈0.05）, both of which reached the highest levels at 50 μg/ml. Compared with normal 16 HBE cells, the expression of IL-17 A induced by silica dust was also significantly decreased in CTGF siRNA-treated 16 HBE cells at 25 and 50 μg/ml. Conclusion Inhibition of CTGF mRNA expression can significantly reduce the level of IL-17 A induced by silica dust in 16 HBE cells, CTGT might play a vital role in regulating the inflammatory and fibrosis responses induced by silica dust.

关 键 词：石英粉尘 人支气管上皮细胞(16HBE) 结缔组织生长因子(CTGF) 白介素17A(IL-17A) 

分 类 号：R994.6[医药卫生—毒理学]