Effects of rapamycin against paraquat-induced pulmonary fibrosis in mice  

雷帕霉素对小鼠百草枯中毒导致肺纤维化的治疗作用(英文)

作　　者：Xue SHAO Meng LI Chong LUO Ying-ying WANG Ying-ying LU Shi FENG Heng LI Xia-bing LANG Yu-cheng WANG Chuan LIN Xiu-jin SHEN Qin ZHOU Hong JIANG Jiang-hua CHEN 

机构地区：[1]Kidney Disease Center, the First Affiliated Hospital, School of Medicine, Zhejiang University

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2015年第1期52-61,共10页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：supported by the National Key Technology R&D Program of China(No.2011BAI10B07);the National Basic Research Program(973)of China(No.2012CB517603);the National High-Tech R&D Program(863)of China(No.2012AA02A512)

摘　　要：Background and aims： Ingestion of paraquat （PQ）, a widely used herbicide, can cause severe toxicity in humans, leading to a poor survival rate and prognosis. One of the main causes of death by PQ is PQ-induced pul- monary fibrosis, for which there are no effective therapies. The aim of this study was to evaluate the effects of ra- pamycin （PAPA） on inhibiting PQ-induced pulmonary fibrosis in mice and to explore its possible mechanisms. Methods： Male C57BL/6J mice were exposed to either saline （control group） or PQ （10 mg/kg body weight, intraper- itoneally; test group）. The test group was divided into four subgroups： a PQ group （PQ-exposed, non-treated）, a PQ＋RAPA group （PQ-exposed, treated with RAPA at I mg/kg intragastrically）, a PQ＋MP group （PQ-exposed, treated with methylprednisolone （MP） at 30 mg/kg intraperitoneally）, and a PQ＋MP＋RAPA group （PQ-exposed, treated with MP at 30 mg/kg intraperitoneally and with PAPA at 1 mg/kg intragastrically）. The survival rate and body weight of all the mice were recorded every day. Three mice in each group were sacrificed at 14 d and the rest at 28 d after intox- ication. Lung tissues were excised and stained with hematoxylin-eosin （H＆E） and Masson＇s trichrome stain for his- topathological analysis. The hydroxyproline （HYP） content in lung tissues was detected using an enzyme-linked im- munosorbent assay （ELISA） kit. The expression of transforming growth factor-β1 （TGF-β1） and α-smooth muscle actin （α-SMA） in lung tissues was detected by immunohistochemical staining and Western blotting. Results： A mice model of PQ-induced pulmonary fibrosis was established. Histological examination of lung tissues showed that PAPA treatment moderated the pathological changes of pulmonary fibrosis, including alveolar collapse and interstitial collagen depo- sition. HYP content in lung tissues increased soon after PQ intoxication but had decreased significantly by the 28th day after PAPA treatment. Immunohistoche目的:评估雷帕霉素(RAPA)对小鼠百草枯(PQ)中毒导致肺纤维化的治疗作用,并初步探讨其作用机制。创新点:首次在小鼠PQ中毒的模型中证明RAPA可明显抑制肺纤维化,且此作用与转化生长因子-β1(TGF-β1)的抑制相关。方法:将C57BL/6J雄鼠分为对照组(腹腔注射生理盐水)和实验组(腹腔注射10 mg/kg PQ)。实验组根据采用的治疗不同,可分为以下四组:PQ组、PQ+RAPA组、PQ+MP(甲强龙)组和PQ+MP+RAPA组。用苏木精-伊红染色法(H&E)和马松(Masson)三色染色法观察肺组织病理结构变化,用酶联免疫吸附测定(ELISA)试剂盒检测肺组织中的羟脯氨酸(HYP)含量,用免疫组化和免疫印迹的方法检测TGF-β1和α-平滑肌肌动蛋白(α-SMA)的表达水平。结论:本实验中小鼠的肺组织病理染色结果显示,RAPA治疗能缓解肺纤维化导致的病理改变(图3)。ELISA实验结果显示,RAPA治疗28天后可显著降低肺组织中HYP的含量(图4)。免疫组化和免疫印迹实验结果显示,RAPA治疗能明显下调肺组织中TGF-β1和α-SMA的高表达(图5和6)。综上所述,RAPA在治疗PQ中毒导致的肺纤维化中有重要价值。

关 键 词：PARAQUAT Pulmonary fibrosis RAPAMYCIN Transforming growth factor-β1 a-Smooth muscle actin METHYLPREDNISOLONE 

分 类 号：R563.9[医药卫生—呼吸系统]