蒿鳖养阴软坚方通过Nrf2/γ-GCS信号转导通路抑制酒精性肝纤维化作用及其机制  

Effects and mechanism of HaoBieYangYinRuanJian prescription on inhibition of alcoholic liver fibrosis through Nrf2/γ-GCS pathway

作　　者：李杉杉 刘悦[1] 唐诗慧 方步武[1] 

机构地区：[1]天津医科大学药理学系,天津300070

出　　处：《天津医科大学学报》2018年第1期14-18,共5页Journal of Tianjin Medical University

摘　　要：目的:研究不同提取方案的蒿鳖养阴软坚方对肝纤维化大鼠肝组织中Nrf2/γ-GCS信号转导通路的影响,探讨不同提取方案的蒿鳖养阴软坚方对大鼠肝纤维化的治疗作用。方法:健康Wistar大鼠随机分为8组:正常组、模型对照组、先水后醇60%提取方案组(0.09 mg·kg^(-1)·d^(-1))、先水后醇95%提取方案低、高剂量组(2.59 g.kg^(-1)·d^(-1)、8.2 g·kg^(-1)·d^(-1))、水提醇沉低、高剂量组(2.59 g·kg^(-1)·d^(-1)、8.2 g·kg^(-1)·d^(-1))及复方鳖甲软肝片组(0.09 mg·kg^(-1)·d^(-1)),每组10只大鼠。酒精性肝纤维化大鼠模型制备成功后,各组按相应药物剂量分别开始治疗,正常组和模型组给予同体积蒸馏水,每日1次,造模10周后处死大鼠,留取标本。酸水解法测定肝组织中胶原蛋白含量;Western blotting检测肝组织Nrf2蛋白与γ-GCS蛋白的表达;光镜观察肝组织免疫组化染色结果。结果:(1)肝组织中羟脯氨酸含量:与正常组比较,模型组大鼠肝组织胶原蛋白含量增高(P<0.01);与模型组比较,先水后醇60%提取方案组、先水后醇中、高剂量组以及复方鳖甲软肝片组大鼠肝组织胶原含量均降低(P<0.05)。(2)肝组织Nrf2蛋白与γ-GCS蛋白的表达:与正常组比较,模型组大鼠肝组织Nrf2蛋白与γ-GCS蛋白表达增高(P<0.01);与模型组比较,各治疗组大鼠肝组织Nrf2蛋白与γ-GCS蛋白表达增高(P<0.01)。(3)肝组织免疫组化染色结果:模型组的Nrf2和γ-GCS蛋白主要在细胞浆中表达,各用药组Nrf2和γ-GCS蛋白均多数在细胞浆和细胞核中表达,汇管区附近也有所表达,但表达量不及胞浆和胞核。结论:提取工艺优化后的蒿鳖养阴软坚方可能通过上调肝组织Nrf2/γ-GCS信号转导通路,增加Nrf2蛋白和γ-GCS蛋白的表达,降低大鼠肝组织肝纤维化程度从而发挥抗肝纤维化的作用。Objective： To investigate the effects of different extraction schemes of Hao Bie Yang Yin Ruan Jian prescription（HBYYRJP） on alcoholic liver disease in rats, and explore the possible mechanisms by upgrading the Nrf2 signaling pathway in liver. Methods：Healthy Wistar rats were randomly divided into 8 groups： normal group, model group, group with low dose of Water followed by 95% ethyl alcohol extraction scheme（2.59 g·kg^-1·d^-1）, group with high dose of Water followed by 95% ethyl alcohol extraction scheme（8.2 g·kg^-1·d^-1）, group with low dose of Water extraction and alcohol precipitation scheme（2.59 g·kg^-1·d^-1）, group with high dose of Water extraction and alcohol precipitation scheme（8.2 g·kg^-1·d^-1）, Powder group（0.09 mg·kg^-1·d^-1） and Compound Biejiaruangan Troche group（0.09 mg.kg～（-1）.d～（-1））. Drugs were given to the corresponding therapeutic groups once a day for ten weeks after alcoholic liver fibrosis models of rats were successfully made. Distilled water was given to the rats of normal and model groups by the same method. After 10 weeks, rats were put to death, and samples were taken. Liver tissues were used for measuring the contents of collagen, Nrf2, γ-GCS protein expression by Western blotting and immunohistochemistry. Results： Compared with the normal group, the contents of collagen were enhanced in model group（P〈0.01）.Compared with the model group, the contents of collagen markedly decreased in group with high dose of Water followed by 95% ethyl alcohol extraction scheme groups（P〈0.01）, Powder group（P〈0.01） and Compound Biejiaruangan Troche group（P〈0.01）. Compared with the normal group, expressions of Nrf2, γ-GCS protein were enhanced in model group（P〈0.01, P〈0.01）. Compared with the model group,expressions of Nrf2, γ-GCS protein markedly enhanced in group with high dose of Water followed by 95% ethyl alcohol extraction scheme（P〈0.01, P〈0.01）, Powder group（P〈0.01, P〈0.01） and Compound Bi

关 键 词：蒿鳖养阴软坚方 酒精性肝纤维化 胶原蛋白 NRF2 Γ-GCS 

分 类 号：R96[医药卫生—药理学]