黄芪甲苷对急性肝衰竭小鼠抗氧化、抗凋亡机制探讨  

Exploration on the Mechanism of Anti-Oxidation and Anti-Apoptosis of AstragalosideⅣin Acute Liver Failure Mice

作　　者：刘丽 周宇 李双杰[3] LIU Li;ZHOU Yu;LI Shuang-jie(Department of Pediatrics,Zhuji People's Hospital of Zhejiang Province,Zhejiang(311800);ICU,Zhuji People's Hospital of Zhejiang Province,Zhejiang(311800);Department of Hepatology,Children's Hospital of Hunan Province,Changsha(410007))

机构地区：[1]诸暨市人民医院儿科,浙江311800 [2]诸暨市人民医院重症监护室,浙江311800 [3]湖南省儿童医院肝病中心,长沙410007

出　　处：《中国中西医结合杂志》2023年第6期722-728,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：浙江省卫生健康科技计划青年创新人才项目(No.2022RC283);诸暨市医药卫生科技计划项目(No.2020YW045)。

摘　　要：目的研究黄芪甲苷(AS-Ⅳ)对急性肝衰竭小鼠转录因子NF-E2相关因子2(Nrf2)/血红素氧化酶-1(HO-1)和B淋巴细胞瘤-2基因(Bcl-2)/Bcl-2相关X蛋白(Bax)表达的影响,并探讨其抗氧化、抗凋亡的作用机制。方法将50只C57BL6雄性小鼠随机分为正常组,模型组,低、中、高剂量AS-Ⅳ组,每组10只。低、中、高剂量AS-Ⅳ组分别给予50、150、500 mg/(kg·d)AS-Ⅳ预处理3天,正常组和模型组仅给予助溶剂预处理,然后正常组腹腔注射生理盐水,余4组分别给予D-GalN/LPS腹腔注射建立急性肝衰竭小鼠模型。留取肝组织病理标本,采用Western Blot和RealTime PCR技术检测各组Nrf2、HO-1、Bcl-2、Bax、半胱氨天冬蛋白酶-3(Caspase-3)蛋白及m RNA的表达水平。结果与正常组比较,模型组小鼠肝组织Nrf2、HO-1、Bcl-2/Bax蛋白及mRNA表达降低(P<0.01),Caspase-3蛋白及m RNA表达增加(P<0.01)。与模型组比较,低剂量AS-Ⅳ组小鼠肝组织Nrf2蛋白及HO-1、Bcl-2/Bax蛋白及mRNA表达增加(P<0.01),Caspase-3蛋白及mRNA表达减低(P<0.01);中剂量AS-Ⅳ组Nrf2 mRNA及HO-1、Bcl-2/Bax蛋白及mRNA表达增加(P<0.05,P<0.01),Caspase-3蛋白及m RNA表达减低(P<0.01);高剂量AS-Ⅳ组Nrf2、HO-1、Bcl-2/Bax蛋白及mRNA表达增高(P<0.01),Caspase-3蛋白及mRNA表达下降(P<0.01)。与低剂量AS-Ⅳ组比较,中、高剂量AS-Ⅳ组HO-1、Bcl-2/Bax蛋白及mRNA表达升高(P<0.05,P<0.01),高剂量AS-Ⅳ组Nrf2蛋白及mRNA表达升高(P<0.05,P<0.01),Caspase-3蛋白及mRNA表达下降(P<0.05,P<0.01);与中剂量ASⅣ组比较,高剂量AS-Ⅳ组HO-1蛋白及m RNA、Bcl-2/Bax蛋白、Nrf2 mRNA表达升高(P<0.05,P<0.01),Caspase-3蛋白及m RNA表达下降(P<0.01)。结论高剂量AS-Ⅳ对急性肝衰竭小鼠有保护作用,其机制可能与通过Nrf2增强HO-1的表达,减轻氧化应激所致急性肝损伤,以及上调Bcl-2的表达,降低Bax、Caspase-3水平,从而抑制肝细胞凋亡有关。Objective To observe the effect of Astragaloside Ⅳ(AS-Ⅳ)on the expression of NF-E2-related factor 2(Nrf2)/heme oxygenase-1(HO)and Bcl-2/Bax in acute liver failure(ALF)mice,and to explore its mechanism of anti-oxidative and anti-apoptotic.Methods Fifty C57BL6 male mice were randomly divided into normal control group,model group,low dose AS-Ⅳ group,medium dose AS-Ⅳgroup,and high dose AS-Ⅳ group,10 mice in each group.Different dosage(50,150,500 mg^(-1)·kg^(-1)·d^(-1))of AS-Ⅳ was administrated by gavage to the three intervention groups for 3 days,at the same time,cosolvent was given to the normal control group and the model group.Then the normal control group was given normal saline by intraperitoneal injection,while the other 4 groups were given D-GalN and LPS by intraperitoneal injection to establish mice models of ALF.Liver histopathological specimens were retained,and the Nrf2,HO-1,Bcl-2,Bax,Caspase-3 protein and m RNA expression in liver tissues were detected by Western Blot and Real-Time PCR.Results Compared with the normal group,the expression of Nrf2,HO-1,and Bcl-2/Bax protein and mRNA in the liver tissue decreased(P<0.01),Caspase-3 protein and mRNA increased(P<0.01).Compared with the model group,the expression of Nrf2 protein,HO-1 and Bcl-2/Bax protein and mRNA increased(P<0.01),Caspase-3 protein and mRNA decreased(P<0.01)in the low dose AS-Ⅳ group.The expression of Nrf2 mRNA,HO-1,and Bcl-2/Bax protein and mRNA increased(P<0.05,P<0.01),Caspase-3 protein and mRNA decreased in the medium dose AS-Ⅳgroup(P<0.01).The expression of Nrf2,HO-1,Bcl-2/Bax protein and mRNA increased(P<0.01),Caspase-3protein and mRNA decreased in the high dose AS-Ⅳ group(P<0.01).Compared with the low dose AS-Ⅳ group,the expression of HO-1 and Bcl-2/Bax protein and mRNA increased(P<0.05,P<0.01)in the medium and high dose AS-Ⅳ group,the expression of Nrf2 protein and mRNA increase(P<0.05,P<0.01),Caspase-3protein and mRNA decreased(P<0.05,P<0.01)in the high dose AS-Ⅳ group.Compared with the medium dose AS-Ⅳgroup

关 键 词：急性肝衰竭 黄芪甲苷 小鼠 抗氧化 抗凋亡 中药 

分 类 号：R285.5[医药卫生—中药学]