基于Keap1/Nrf2信号通路探讨丹葛解酲汤对酒精性肝病大鼠模型的作用机制  

Mechanism of action of Dange Jiecheng decoction in a rat model of alcoholic liver disease based on the Kelch-like ECH-associated protein 1/nuclear factor erythroid 2-related factor 2 signaling pathway

作　　者：王铭 徐建虎[1,2] 马丽 WANG Ming;XU Jianhu;MA Li(School of Traditional Chinese Medicine,Ningxia Medical University,Yinchuan 750004,China;Key Laboratory of Ningxia Minority Medicine Modernization,Ministry of Education,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学中医学院,银川750004 [2]宁夏少数民族医药现代化教育部重点实验室,银川750004

出　　处：《临床肝胆病杂志》2023年第5期1119-1125,共7页Journal of Clinical Hepatology

基　　金：宁夏自然科学基金(2021AAC03146);宁夏医科大学校级特殊人才启动项目(XT2020027)。

摘　　要：目的 观察丹葛解酲汤对酒精性肝病(ALD)大鼠模型的治疗作用,探讨丹葛解酲汤抗氧化应激的作用机制。方法 将96只SD大鼠随机分为空白组(n=13)和ALD组(n=83),ALD组采用白酒灌胃法建立ALD模型。随后将存活ALD组大鼠随机分为模型组、丹葛解酲汤高剂量组(24 g/kg)、丹葛解酲汤低剂量组(6 g/kg)及益肝灵片组(21 mg/kg),每组各17只,空白组和模型组给予生理盐水灌胃,其余组分别灌胃给予相应的药物,共干预4周。采用HE染色法观察大鼠肝组织的病理变化;Western Blot法检测肝脏组织Kelch样ECH相关蛋白1(Keap1)、核转录因子E2相关因子2(Nrf2)、血红素氧合酶1(HO-1)的蛋白含量;实时荧光定量PCR检测肝组织Keap1、HO-1 mRNA表达水平。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 与空白组相比,模型组肝细胞排列紊乱,出现肝细胞坏死,有大量炎性细胞浸润,并伴有大量的脂滴空泡;Keap1蛋白、Keap1 mRNA表达水平均明显上升(P值均<0.05),Nrf2蛋白、HO-1蛋白、HO-1 mRNA表达水平均明显下降(P值均<0.05)。与模型组相比,丹葛解酲汤高剂量组、丹葛解酲汤低剂量组及益肝灵片组肝细胞排列变整齐,坏死及炎性细胞减少,偶见或无脂滴空泡;Keap1蛋白、Keap1 mRNA表达水平显著降低(P值均<0.05),Nrf2蛋白、HO-1蛋白、HO-1 mRNA表达水平显著上升(P值均<0.05)。结论 丹葛解酲汤可能通过调节Keap1/Nrf2信号通路,促使Nrf2入核,上调HO-1的表达,减轻氧化应激反应,进而对ALD大鼠发挥保护作用。Objective To investigate the therapeutic effect of Dange Jiecheng decoction on a rat model of alcoholic liver disease(ALD)and the anti-oxidative stress mechanism of Dange Jiecheng decoction.Methods A total of 96 Sprague-Dawley rats were randomly divided into blank group with 13 rats and ALD group with 83 rats,and the rats in the ALD group were given liquor by gavage to establish a model of ALD.Then the ALD group was randomly divided into model group,high-dose Dange Jiecheng decoction group(24 g/kg),low-dose Dange Jiecheng decoction group(6 g/kg),and Yiganling tablet group(21 mg/kg),with 17 rats in each group.The rats in the blank group and the model group were given normal saline by gavage,and those in the other groups were given corresponding drugs by gavage,for 4 consecutive weeks.HE staining was used to observe the pathological changes of liver tissue;Western blot was used to measure the contents of Kelch-like ECH-associated protein 1(Keap1),nuclear factor erythroid 2-related factor 2(Nrf2),and heme oxygenase-1(HO-1)in liver tissue;quantitative real-time PCR was used to measure the mRNA expression levels of Keap1 and HO-1 in liver tissue.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results Compared with the blank group,the model group had disordered arrangement of hepatocytes with necrosis,massive inflammatory cell infiltration,and a large number of lipid droplet vacuoles,significant increases in the protein and mRNA expression levels of Keap1(P<0.05),and significant reductions in the protein expression levels of Nrf2 and HO-1 and the mRNA expression level of HO-1(P<0.05).Compared with the model group,the high-and low-dose Dange Jiecheng decoction groups and the Yiganling tablet group had ordered arrangement of hepatocytes,reductions in hepatocyte necrosis and inflammatory cells,and occasional lipid droplet vacuoles,as well as significant reductions in the protein

关 键 词：肝疾病 酒精性 Kelch样ECH相关蛋白1 NF-E2相关因子2 丹葛解酲汤 大鼠 Sprague-Dawley 

分 类 号：R285.5[医药卫生—中药学]