结缔组织生长因子在百草枯致肺纤维化中的作用  

Fhe role of connective tissue growth factor in pulmonary fibrosis induced by paraquat

作　　者：符红娜 孙兆瑞 葛鑫[1] 曹丽萍[1] 张炜[1] 杨志洲 聂时南[1] Fu Hongna,Sun Zhaorui, Ge Xin, Cao Liping, Zhang Wei, Yang Zhizhou, Nie Shinan(Department of Emergency Medicine, Nanjing General Hospital, PLA, Nanjing 210002, China)

机构地区：[1]中国人民解放军南京总医院急救医学科,南京210002

出　　处：《中华急诊医学杂志》2018年第11期1198-1204,共7页Chinese Journal of Emergency Medicine

基　　金：国家自然科学基金青年基金（81701894,81401583）;江苏省社会发展面上项目（BE2017720）;江苏省科教强卫工程青年医学人才项目（QNRC2016908）

摘　　要：目的研究结缔组织生长因子（connective tissue growth factor,CTGF）在百草枯中毒致肺纤维化发生发展中的作用。方法于南京大学医学院实验室完成实验，48只雄性SD大鼠随机（随机数字法）分为两组，实验组接受单次腹腔注射百草枯（30mg／kg），对照组注射等量生理盐水。分别在第7、14和28天后取下大鼠左肺，进行HE染色、Masson染色及测定羟脯氨酸含量来检测胶原蛋白的表达，评估肺纤维化发生及严重程度，Western-blot及免疫组化检测CTGF的表达。将MRC．5人肺成纤维细胞暴露于不同浓度的百草枯（50-500μmol／L）3d，Western—blot观察CTGF表达情况；用不同浓度CTGF处理MRC-5细胞，分别用CCK-8、Transwell技术观察其增殖及迁移，并用Western—blot验证肌成纤维细胞分化。采用SPSS18．0进行数据统计。结果PQ给药后2周，肺组织切片显示肺泡壁显著增厚并出现具有成纤维特征的间质细胞的积聚。Masson染色显示胶原沉积成斑片状分布，提示肺纤维化形成。Western—Blot和免疫组化证明实验组大鼠CTGF表达显著上调（P〈0．01）。同样，不同浓度PQ处理的MRC-5细胞CTGF表达增加（P〈0．01），具有剂量依赖性，浓度越高CTGF表达增加。此外，CTGF可促进MRC-5细胞的增殖和迁移（P〈0．01），并诱导其分化为肌成纤维细胞。结论PQ可引起CTGF的表达增加，诱导肺成纤维细胞的增殖、迁移和分化。CTGF可能在PQ诱导的肺纤维化中起重要作用，有望成为潜在的药物靶点。Objective To examine the role of connective tissue growth factor （CTGF） in a rat model of paraquat （PQ）-induced pulmonary fibrosis. Methods In the laboratory of Medical School of Nanjing University, 48 adult male Sprague-Dawley （SD） rats were randomly（random number） divided into two groups. The experimental group received a single intraperitoneal injection of PQ （30 mg/kg）, while the control group received an equivalent volume of sterile saline. The rats were then sacrificed and the left lungs were collected on 7th, 14th and 28th day, respectively. HE staining, Masson staining and hydroxyproline content analysis were used to determine the quantity of collagen, and the fibrosis process and severity were evaluated. Western blotting and immunohistochemical staining was used to detect the expression of CTGF. MRC-5 human lung fibroblasts were exposed to different concentrations of PQ （50- 500 μmol/L） for 3 days. The proliferation, migration and differentiation of MRC-5 cells were investigated by CCKS, Transwell and Western blotting after exposing with various concentrations of CTGF （50-200 ng/mL）. Data was analyzed with SPSS18.0. Results At 2 weeks after PQ administration, lung tissue sections exhibited a marked thickening of the alveolar walls with an accumulation of interstitial cells with a fibroblastic appearance. Masson staining revealed a patchy distribution of collagen deposition, indicating pulmonary fibrogenesis. Western-blott and immunohistochemical staining demonstrated that CTGF expression was significantly upregulated in the PQ-treated group （P〈0.01）. Similarly, CTGF expression in PQ-treated MRC-5 cells was increased in a dose-dependent manner （P〈0.01）. In addition, CTGF promoted the proliferation and migration of MRC-5 cells （P〈0.01）, and induced the differentiation of fibrobtasts into myofibroblasts. Conclusions These findings demonstrate that PQ can increase CTGF expression, which may be important in PQ-induced pulmonary fibrogenesis. Therefore, this growt

关 键 词：百草枯 肺纤维化 结缔组织生长因子 肌成纤维细胞分化 治疗靶点 

分 类 号：R563[医药卫生—呼吸系统] R595.4[医药卫生—内科学]