IL-17 A对系统性硬化病小鼠模型皮肤和肺部纤维化病变的作用  

Effects of IL-17A on fibrosis of skin and lung in a mouse model of systemic sclerosis

作　　者：黄舒柠 雷玲[1] 赵铖[1] 王旭 文静[1] 覃芳[1] Huang Shu-ning Lei Ling Zhao Cheng Wang Xu Wen Jing Qin Fang(Department of Rheumatology and Immunology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China Department of Rheumatology and Immunology, People＇s Hospital of Zhengzhou, Zhengzhou 450003, China)

机构地区：[1]广西医科大学第一附属医院风湿免疫科,南宁530021 [2]郑州市人民医院风湿免疫科,450003

出　　处：《中华微生物学和免疫学杂志》2017年第2期105-111,共7页Chinese Journal of Microbiology and Immunology

基　　金：广西自然科学基金项目（2016GXNSFAA380175）

摘　　要：目的 研究白细胞介素（IL）-17A在博莱霉素（BLM）致系统性硬化病（SSc）小鼠模型中的表达及对皮肤、肺部炎症和纤维化病变的作用.方法 24只雌性BALB/c小鼠随机分为4组,包括正常对照组（小鼠背部皮下注射磷酸盐缓冲液）、模型组（小鼠背部皮下注射BLM）、抗体组（BLM＋抗IL-17A单克隆抗体）、同型对照组（BLM＋同型对照）.观察各组小鼠背部注射部位皮肤和肺部的病理改变、炎症和纤维化评分,免疫组化和实时荧光定量PCR（RT-PCR）检测小鼠皮肤和肺部的IL-17A、TGF-β1、Ⅰ型胶原mRNA的表达.体外培养小鼠肺成纤维细胞（FB）,分别加入IL-17A细胞因子和单克隆抗体,检测各组细胞Ⅰ型胶原mRNA、TGF-β1 mRNA的表达,酶联免疫吸附试验（ELISA）检测培养液上清IL-6、TGF-β1的水平.结果 （1）抗体组小鼠皮肤厚度、皮肤炎症、肺组织炎症和纤维化评分明显低于模型组及同型对照组（P＆lt;0.05）.（2）免疫组化提示模型组IL-17A在皮肤和肺组织的表达明显高于正常对照组,抗体组皮肤和肺组织IL-17A蛋白的表达均低于模型组和同型对照组（P＆lt;0.05）.（3）模型组IL-17A mRNA在皮肤和肺组织的表达明显高于正常对照组,抗体组小鼠皮肤和肺组织中IL-17A、TGF-β1、Ⅰ型胶原mRNA的表达均低于模型组和同型对照组（P＆lt;0.05）.（4）体外细胞培养SSc小鼠肺FB,IL-17A单克隆抗体阻断组TGF-β1、Ⅰ型胶原mRNA水平、 细胞上清液IL-6、TGF-β1水平均低于IL-17A细胞因子刺激组（P＆lt;0.05）,而均高于对照组（P＆lt;0.05）.结论 IL-17A可促进SSc小鼠模型皮肤和肺脏炎症和纤维化的发展,阻断IL-17A可能通过抑制TGF-β1、IL-6和Ⅰ型胶原的产生进而抑制SSc纤维化病变.Objective To analyze the expression of interleukin （ IL）-17A in a mouse model of bleomycin （ BLM）-induced systemic sclerosis （ SSc） and to evaluate its effects on inflammation and fibrosis in skin and lung tissues. Methods Twenty-four female BALB/c mice were randomly divided into four groups：normal control group （ mice were subcutaneously injected with phosphate buffer ） , model group （subcutaneously injected with BLM）, antibody group （injected with BLM ＋ IL-17A monoclonal antibody）, homotypic control group （ injected with BLM ＋ isotype control） . Pathological changes in skin and lung tis-sues of those mice were observed;inflammatory and fibrotic scores were assessed. Immunohistochemistry and real-time fluorescent quantitative PCR （ RT-PCR） were used to detect the expression of IL-17A, TGF-β1 and typeⅠ collagen in skin and lung tissues of those mice at mRNA level. Mouse lung fibroblasts （ FB） de-rived from the mice of model group were cultured in vitro and then were cultured with IL-17A cytokines with or without the interference of monoclonal antibodies. Expression of typeⅠ collagen and TGF-β1 at mRNA level and levels of IL-6 and TGF-β1 in the culture supernatants were detected by RT-PCR and enzyme-linkedimmunosorbent assay （ ELISA） , respectively. Results Compared with the mice of model and homotypic control groups, those of the antibody group showed mild skin thickening, skin inflammation and lung inflam-mation as well as lower fibrosis scores （P＆lt;0. 05）. The expression of IL-17A at both protein and mRNA lev-els and the expression of TGF-β1 and collagen typeⅠat mRNA level in skin and lung tissues of mice of the antibody group were significantly lower than those of the model and homotypic control group （P＆lt;0. 05）. Re-sults of the in vitro cell culture of SSc mice-derived lung FB with IL-17A showed that the expression of TGF-β1 and typeⅠ collagen at mRNA level and the levels of IL-6 and TGF-β1 in the culture supernatants were decreased wi

关 键 词：系统性硬化病 抗白细胞介素17A单克隆抗体 转化生长因子Β1 Ⅰ型胶原 TGF-β1 

分 类 号：R-332[医药卫生] R593.2