熊去氧胆酸通过调控TGF-β1/Smads信号通路对博来霉素诱导大鼠肺纤维化的影响  

Ursodeoxycholic Acid Regulates TGF-β1/Smads Signaling Pathway to Treat Bleomycin-Induced Pulmonary Fibrosis in Rats

作　　者：张亮[1] 席瑞[2] 臧凝子[2] ZHANG Liang;XI Rui;ZANG Ning-zi(Liaoning University of Traditional Chinese Medicine,Shenyang Liaoning 110847;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang Liaoning 110032)

机构地区：[1]辽宁中医药大学,辽宁沈阳110032 [2]辽宁中医药大学附属医院,辽宁沈阳110032

出　　处：《世界中西医结合杂志》2023年第2期271-276,共6页World Journal of Integrated Traditional and Western Medicine

基　　金：国家中医药管理局第四批全国中医(西学中)优秀人才研修项目(No.国中医药人教发[2019]13号);辽宁省教育厅科学技术研究项目(L201913)。

摘　　要：目的基于转化生长因子-β1(Transforming growth factor beta1,TGF-β1)/Smads信号通路探讨熊去氧胆酸(Ursodeoxycholic Acid,UDCA)调控通过博来霉素诱导的大鼠肺纤维化的作用机制。方法选取100只成年健康雄性Wistar大鼠作为研究对象,采用随机数表法进行分组,分为空白组、模型组、低剂量熊去氧胆酸+博来霉素组、中剂量熊去氧胆酸+博来霉素组、高剂量熊去氧胆酸+博来霉素组,每组各20只。除空白组大鼠外,其余各组均给予大鼠单次气管内滴注博来霉素来建立实验性的肺纤维化动物模型。于造模当天开始给予低剂量熊去氧胆酸+博来霉素组、中剂量熊去氧胆酸+博来霉素组、高剂量熊去氧胆酸+博来霉素组大鼠熊去氧胆酸(剂量分别为50 mg/(kg·d)、100 mg/(kg·d)、200 mg/(kg·d))1次/d灌胃。实验第1、7、14、21天所有各组分别抽取4只大鼠,采用酶联免疫吸附剂测定法(Enzyme-Linked ImmunoSorbent Assay,ELISA)检测大鼠血清中TGF-β1的表达水平。第21天采用苏木素-伊红染色(HE染色)法观察大鼠肺组织病理变化,采用Masson染色观察胶原沉积情况,采用Western Blotting方法观察各组大鼠肺组织中TGF-β1、p-Smad2、Smad2、p-Smad3、Smad3、α-SMA蛋白表达的情况。结果第7天,与模型组比较,中剂量熊去氧胆酸+博来霉素组大鼠血清TGF-β1水平明显降低(P<0.01);与低剂量熊去氧胆酸+博来霉素组比较,中剂量熊去氧胆酸+博来霉素组、高剂量熊去氧胆酸+博来霉素组大鼠血清TGF-β1水平明显降低(P<0.01,P<0.05)。第14、21天,与模型组比较,低剂量熊去氧胆酸+博来霉素组、中剂量熊去氧胆酸+博来霉素组、高剂量熊去氧胆酸+博来霉素组大鼠血清TGF-β1水平明显降低(P<0.01)。第14天,与低剂量熊去氧胆酸+博来霉素组、中剂量熊去氧胆酸+博来霉素组比较,高剂量熊去氧胆酸+博来霉素组大鼠血清TGF-β1水平明显降低(P<0.05)。第21天,与低剂量熊去氧Objective To explore the mechanism of ursodeoxycholic acid(UDCA)in the treatment of bleomycin-induced pulmonary fibrosis in rats via the transforming growth factor-β1(TGF-β1)/Smads signaling pathway.Methods One hundred adult healthy male Wistar rats were randomly assigned into a blank group,a model group,and low-dose,medium-dose and high-dose UDCA+bleomycin groups,with 20 rats in each group.For the rats in other groups exceptthe blank group,a single dose of bleomycin was administrated by intratracheal infusion to establish the model of pulmonary fibrosis.The rats in the low-,medium-,and high-dose UDCA+bleomycin groups had been administrated with 50,100,and 200 mg/(kg·d)UDCA respectively by gavage once a day since the day of modeling.On days 1,7,14,and 21,4 rats were selected from each group for measurement of the serum level of TGF-β1 by enzyme-linked immunosorbent assay(ELISA).On day 21,the pathological changes in the lung tissue were observed by hematoxylin-eosin(HE)staining.Meanwhile,the collagen deposition was observed by Masson staining,and the protein levels of TGF-β1,p-Smad2,Smad2,p-Smad3,Smad3,andα-SMA in the lung tissue were measured by Western blotting.Results On day 7,compared with model group,serum TGF-β1 level in medium-dose UDCA+bleomycin group was significantly decreased(P<0.01);Compared with low-dose UDCA+bleomycin group,serum TGF-β1 level in medium-dose UDCA+bleomycin group and high-dose UDCA+bleomycin group was significantly decreased(P<0.01,P<0.05,respectively).On days 14 and 21,compared with the model group,the administration with UDCA at all the three doses groups lowered the serum level of TGF-β1(P<0.01).Moreover,the low-and medium-dose UDCA+bleomycin groups had higher serum level of TGF-β1 than the high-dose UDCA+bleomycin group(P<0.05 on day 14 and P<0.01 on day 21,respectively).Compared with model group,the administration with UDCA at all the three doses groups down-regulated the protein level of TGF-β1 and decreased the p-Smad2/Smad2 ratio in the lung tissue(P<0.01);the admini

关 键 词：肺纤维化 熊去氧胆酸 TGF-β1/Smads信号通路 博来霉素 

分 类 号：R285.5[医药卫生—中药学]