富勒烯C60对博来霉素诱导的小鼠肺纤维化的影响  

Effect of water-soluble C6o fullerenes on pulmonary fibrosis induced by bleomycin in mice

作　　者：董润 刘敏[3] 黄骁舾[2] 刘正[2] 姜丁源[4] 肖慧娟[1] 代华平[4] Dong Run Liu Min Huang Xiaoxi Liu Zheng Jiang Dingyuan Xiao Huijuan Dai Huaping(Department of Respirntory and Critical Care Medicine, Beijing Chaoyang Hospital, Capital Medical University, Beijing Key Laboratory of Respiratory and Pulmonary Circulation Disorders, Beijing Institute of Respiratory Medicine, Beijing 100020, China)

机构地区：[1]首都医科大学附属北京朝阳医院呼吸与危重症医学科北京呼吸疾病研究所北京市呼吸与肺循环疾病重点实验室,100020 [2]首都医科大学附属北京朝阳医院医学研究中心,100020 [3]中日友好医院影像诊断科 [4]中日友好医院呼吸与危重症医学科

出　　处：《中华医学杂志》2017年第22期1740-1744,共5页National Medical Journal of China

基　　金：国家自然科学基金（81270123,81470258）

摘　　要：目的观察富勒烯C60对博来霉素诱导的小鼠肺纤维化的影响。方法健康C57BL/6J小鼠20只，按随机数字表法随机分为正常对照组、博来霉素组、高剂量C60组、低剂量C60组各5只。博来霉素组、低剂量C60组及高剂量C60组通过气管内注射博来霉素诱导小鼠肺纤维化模型，正常对照组则以生理盐水代替博来霉素。低剂量C60组、高剂量C60组于注射博来霉素前1天开始，每天通过腹腔注射分别给予小鼠1 mg/kg、10 mg/kg水溶性富勒烯C60，正常对照组及博来霉素组则给予等体积生理盐水。观察14 d小鼠的生存率、体质量变化；通过HE及Masson染色，根据Ashoft评分评价肺纤维化程度；通过碱水解法测定肺组织羟脯氨酸含量；采用酶联免疫吸附（ELISA）法测定肺组织、血浆以及支气管肺泡灌洗液（BALF）中转化生长因子β1（TGF－β1）、肿瘤坏死因子α（TNF－α）的水平；运用DCFH－DA探针检测活性氧的含量[以荧光强度/微克蛋白（OD/μg）表示]。结果富勒烯C60干预后小鼠体质量下降减缓；低剂量C60组、高剂量C60组病理形态学上纤维化Ashoft评分均显著低于博来霉素组[（4.08±0.52）、（3.00±0.41）比（6.75±0.75）分，均P〈0.01]；低剂量C60组、高剂量C60组肺组织羟脯氨酸含量均显著低于博来霉素组[（0.36±0.06）、（0.35±0.08）比（0.55±0.16）μg/mg，均P〈0.05]；高剂量C60组支气管肺泡灌洗液（BALF）、肺组织中TGF－β1含量均低于博来霉素组，但差异均无统计学意义[（9.38±5.32）比（23.60±8.96）pg/ml、（2.89±0.35）比（6.44±2.95）pg/mg，均P〉0.05]，高剂量C60组血浆、BALF及肺组织中TNF－α含量均显著低于博来霉素组[（4.56±0.73）比（7.21±2.26）pg/ml、（34.58±23.30）比（151.00±27.34）pg/ml、（22.99±5.83）比（122.90±22.04）pg/mg，均P〈0.05]；并且，高剂量C60组肺组织中活性氧含量显著低于博来霉素组[（19.68±ObjectiveTo evaluate the role of water-soluble C60 fullerenes in mice model of lung injury and fibrosis that induced by bleomycin.MethodsA total of 20 healthy C57BL/6J mice were randomly divided into normal control group, bleomycin group, high dose C60 group, low dose C60 group, each group with 5 mice. Mice were induced pulmonary fibrosis by intratracheal injection of bleomycin except the normal control group, which was induced by saline instead. In low dose C60 group and high dose C60 group, 1 mg·kg-1·d-1 and 10 mg·kg-1·d-1 water-soluble C60 fullerenes was injected into mice intraperitoneally every day, which began from one day before intratracheal instillation of bleomycin until the end of observation. Saline was given to mice in the same way in normal control and bleomycin group. This study investigated the variation of weight and survival rate of mice for 14 d. HE-staining and Masson′s trichrome staining were used to assess the severity of fibrosis according to the method proposed by Ashcroft at 14th day. Total lung collagen content was determined by hydroxyproline assay. The changes of transforming growth factor-β1 （TGF-β1） and tumor necrosis factor α （TNF-α） in plasma, bronchial alveolar lavage fluid （BALF） and lung tissue were measured by enzyme-linked immunosorbent assay （ELISA）. And, the amount of reactive oxygen species （ROS） was tested by 2, 7-dichlorofuorescin diacetate （DCFH-DA）, and determined by the ratio of fluorescence intensity and protein content （OD/μg）.ResultsC60 can protect mice that injured by bleomycin from weight loss. According the method proposed by Ashcroft et al.HE and Masson′s trichrome staining showed that collagen deposition in lung tissue were markedly attenuated in C60 （1 mg·kg-1·d-1 and 10 mg·kg-1·d-1） treated mice compared with bleomycin model mice[（4.08±0.52）, （3.00±0.41） vs （6.75±0.75） points, both P〈0.01]. In low dose C60 group and high dose C60 group, the content of hydroxyproline in lung tissue were signific

关 键 词：肺纤维化 博来霉素 富勒烯 小鼠 

分 类 号：R563[医药卫生—呼吸系统]