基于Nrf2/HO-1信号通路探讨四逆散对胆汁淤积性肝炎大鼠氧化应激反应的影响  

Effect of Sinisan on Oxidative Stress in Cholestatic Hepatitis Rats Based on Nrf2/HO-1 Axis

作　　者：曹丹 陈祺 陈小橹 陈林珍 王海燕 郝巨辉 张威[5] 马志强[1] CAO Dan;CHEN Qi;CHEN Xiaolu;CHEN Linzhen;WANG Haiyan;HAO Juhui;ZHANG Wei;MA Zhiqiang(Beijing University of Chinese Medicine,Beijing Key Laboratory for Quality Evaluation of Traditional Chinese Medicine,Beijing 102488,China;Pharmaceutical Factory of Beijing Tongrentang Technology Development Co.,Ltd.,Beijing 100079,China;Reproductive and Genetic Medical Center,Dalian Women and Children’s Medical Group,Dalian 116000,China;School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;The First Affiliated Hospital of Harbin Medical University,Harbin 150001,China)

机构地区：[1]北京中医药大学中药品质评价北京市重点实验室,北京102488 [2]北京同仁堂科技发展股份有限公司制药厂,北京100079 [3]大连市妇女儿童医疗中心(集团),生殖与遗传医学中心,辽宁大连116000 [4]北京中医药大学中药学院,北京102488 [5]哈尔滨医科大学附属第一医院,哈尔滨150001

出　　处：《中国实验方剂学杂志》2024年第12期1-7,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82374181);北京中医药大学2023年度基本科研业务费揭榜挂帅项目(2023-YJB-JBZD-036)。

摘　　要：目的:基于核因子E2相关因子2(Nrf2)/血红素氧合酶-1(HO-1)信号通路探讨四逆散对胆汁淤积性肝炎大鼠肝脏氧化应激损伤的影响及其作用机制。方法:30只6周雄性SD大鼠,随机分为正常组、模型组、四逆散低、高剂量组(2.5、5.0 g·kg^(-1))、熊去氧胆酸(UDCA,63 mg·kg^(-1))组,每组6只。各组大鼠连续给药7 d,第5天除对照组给予10 m L·kg^(-1)橄榄油外,其余各组均给予80 mg·kg^(-1)的α-萘异硫氰酸酯(ANIT);采用酶联免疫吸附测定法(ELISA)检测血清中胆汁淤积相关生化指标水平,以及肝组织中抗氧化因子的含量;通过苏木素-伊红(HE)染色观察肝组织病理变化;采用实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测肝组织中Nrf2、HO-1和醌氧化还原酶1(NQO1) mRNA与蛋白的相对表达量。结果:与正常组比较,模型组血清各胆汁淤积生化指标水平、肝组织中抗氧化应激因子含量均显著升高(P<0.01),模型组肝细胞排列紊乱、门管区有明显的瘀血性坏死,炎性细胞浸润,小叶间胆管破坏等病理学改变;模型组大鼠肝组织中Nrf2、HO-1和NQO1 mRNA与蛋白相对表达水平均明显下调(P<0.05,P<0.01)。与模型组比较,四逆散高剂量组的血清各胆汁淤积生化指标水平、肝组织中抗氧化应激因子含量均显著降低(P<0.01);肝病理损伤较模型组有明显改善,表现为坏死面积明显缩小,炎性细胞浸润显著减少,小叶间静脉有少量瘀血;肝组织中Nrf2、HO-1和NQO1 mRNA与蛋白相对表达水平均明显上调(P<0.05,P<0.01)。结论:四逆散能显著改善肝内胆汁淤积性肝炎大鼠肝损伤,其机制可能与Nrf2/HO-1信号通路介导的抑制氧化应激反应有关。Objective:Based on the nuclear factor erythroid 2 related factor 2(Nrf2)/heme oxygenase 1(HO-1)axis,to explore the effect of Sinisan(SNS)on liver oxidative stress injury in cholestatic hepatitis rats and its mechanism.Method:Thirty 6-week-old male SD rats were randomly divided into control group,model group,low and high dose group of SNS(2.5,5 g·kg-1,respectively)and ursodeoxycholic acid(UDCA,63 mg·kg-1)group,with six rats in each group.Rats except control group were administrated for 7 consecutive days,on the 5th day,the control group was given olive oil(10 mL·kg-1),and the other groups were given alpha-naphthalene isothiocyanate(ANIT,80 mg·kg-1).The serum cholestasis biochemical indicators and antioxidant factors in rat liver were detected by enzyme-linked immunosorbent assay(ELISA).Hematoxylin-eosin(HE)staining was used to observe the pathological changes of liver tissues.The relative gene and protein expressions of Nrf2,HO-1 and quinone oxidoreductase 1(NQO1)in liver tissues were detected by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)and western blot.Result:Compared with control group,the model group showed a significant increase in the levels of serum biochemical indicators and antioxidant factors in liver tissues(P<0.01).There were obvious pathological changes in the model group following disordered arrangement of hepatocytes,obvious congestion and necrosis in portal area,infiltration of inflammatory cells and destruction of interlobular bile duct.The relative gene and protein levels of Nrf2,HO-1 and NQO1 in liver were significantly down-regulated in model group(P<0.05,P<0.01).Compared with model group,the low-and high-dose groups of SNS showed a significant decrease in the levels of serum biochemical indicators and antioxidant factors in liver tissues(P<0.01),the pathological liver injury were obviously improved following with the necrotic area reduced,the infiltration of inflammatory cells decreased and small amount of extravasated blood in interlobular vein.The relati

关 键 词：四逆散 胆汁淤积性肝炎 氧化应激 核因子E2相关因子2(Nrf2) 血红素氧合酶-1(HO-1) 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33