大黄素通过抑制TGF-β1/ADAMTS-1通路的活性减轻大鼠肺纤维化  

Emodin alleviates pulmonary fibrosis through inactivation of TGF-β1/ADAMTS-1 signaling pathway in rats

作　　者：刘理静 钱红 肖华[2] 贺兼斌[3] 谢茂峰[3] 王在岩[2] 龙兴云[2] LIU Lijing QIAN Hong XIAO Hua HE Jianbin XIE Maofeng WANG Zaiyan LONG Xingyun(School of Clinical Medicine, Hunan University of Medicine, Huaihua 418000 Department of Respiration, First Affiliated Hospital, University of South China, Hengyang 421001 Department of Respiration, Huaihua First People＇s Hospital, Huaihua 418000, China)

机构地区：[1]湖南医药学院临床医学院,湖南怀化418000 [2]南华大学附属第一医院呼吸内科,湖南衡阳421001 [3]怀化市第一人民医院呼吸内科,湖南怀化418000

出　　处：《细胞与分子免疫学杂志》2016年第10期1342-1346,1351,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：湖南省教育厅优秀青年科研项目(14B142);怀化市科技计划项目(2015F3101)

摘　　要：目的探讨转化生长因子β1(TGF-β1)/含1型血小板结合蛋白基序的解聚蛋白样金属蛋白酶(ADAMTS-1)信号通路在大黄素抗肺纤维化中的作用。方法按随机数字表法将60只雄性SD大鼠分为6组:正常对照组、假手术组、模型组、大黄素低剂量组(20 mg/kg)、大黄素高剂量组(80 mg/kg)和泼尼松组(5 mg/kg),每组10只大鼠。假手术组大鼠气管内注入生理盐水,而后4组大鼠气管内注入博莱霉素A5建立肺纤维化模型,次日起,大黄素低、高剂量组大鼠分别以20 mg/kg、80 mg/kg大黄素2 m L灌胃,泼尼松组予5 mg/kg醋酸泼尼松2 m L灌胃,其余3组则以生理盐水2 m L灌胃。造模后第28天处死大鼠,取血并分离肺组织。常规HE和Masson染色观察肺组织病理改变,荧光实时定量PCR检测各组大鼠肺组织中TGF-β1、ADAMTS-1、1型胶原蛋白(Col1)、Col3 mRNA的水平,Western blot法检测肺组织TGF-β1、ADAMTS-1、Col1、Col3蛋白水平,ELISA测定血清中1型前胶原蛋白羧基端前肽(P1CP)和3型前胶原蛋白氨基端前肽(P3NP)水平。结果与模型组比较,各药物处理组肺泡炎及肺纤维化程度明显减轻。与正常对照组、假手术组比较,模型组肺组织TGF-β1、Col1、Col3 mRNA与蛋白表达水平以及血清P1CP、P3NP浓度升高,而肺组织ADAMTS-1 mRNA与蛋白表达水平降低。与模型组相比,给予低、高剂量大黄素或泼尼松处理后,肺组织TGF-β1、Col1、Col3 mRNA与蛋白表达水平及血清P1CP、P3NP浓度则下调,肺组织ADAMTS-1 mRNA与蛋白表达水平上调。与大黄素低剂量组相比,大黄素高剂量组和泼尼松组上述指标明显改善,但后二者相比无明显差异。结论大黄素抑制TGF-β1/ADAMTS-1途径的活性引起肺组织Col1、Col3降解增多,是其抗肺纤维化的重要机制。Objective To explore the role of transforming growth factor-β1（ TGF-β1） /a disintegrin-like and metal oproteinase with thrombospondin type 1 motif（ ADAMTS-1） signaling pathway in emodin＇s anti-pulmonary fibrosis. Methods Sixty SD rats were randomly divided into 6 groups： normal control group,sham-operated group,model group,low-dose emodin intervention group（ 20 mg / kg）,high-dose emodin intervention group（ 80 mg / kg） and prednisone group（ 5 mg / kg）. Each group included 10 animals. Rats in the latter 4 groups were intratracheally injected with bleomycin A5 to induce pulmonary fibrosis,whereas bleomycin A5 was replaced by normal saline in sham-operated group. From the second day,rats in the low-and high-dose emodin intervention groups were intragastrical y treated with 2 m L of 20 and 80 mg / kg emodin,respectively.Rats in the prednisone group were intragastrically administrated with 2 m L of 5 mg / kg prednisone acetate. However,rats in the normal control and sham-operated and model groups were treated with 2 m L of normal saline. All rats were sacrificed on day 28 after modeling. Subsequently,blood and pulmonary tissue specimen were taken. The pathological changes of pulmonary tissues were observed using routine HE and Masson staining. The expressions of TGF-β1,ADAMTS-1,col agen type 1（ Col1）and Col3 in pulmonary tissues were measured by quantitative real-time PCR and Western blotting. Serum levels of procollagentype 1 carboxy terminal propeptide（ P1CP） and procollagen type 3 aminoterminal propeptide（ P3NP） were detected by ELISA. Results Compare with the model group,the alveolitis and pulmonary fibrosis extent in each drug-treated group were significantly alleviated. In comparison with normal control group or sham-operated group,the mRNA and protein levels of TGF-β1,Col1 and Col3 in pulmonary tissues and the serum levels of P1 CP and P3 NP increased,but the mRNA and protein levels of ADAMTS-1 decreased in model group. After treatment with low-and high-dose emodin or pred

关 键 词：大黄素 肺纤维化 转化生长因子β1 含1型血小板结合蛋白基序的解聚蛋白样金属蛋白酶 1型胶原蛋白 3型胶原蛋白 

分 类 号：R563[医药卫生—呼吸系统] R392.12[医药卫生—内科学]