苦参总黄酮调控Nrf2/HO-1通路对肝纤维化大鼠的机制研究  

Mechanism of total flavonoids of Sophora flavescens regulating Nrf2/HO-1 signaling pathway in rats with liver fibrosis

作　　者：郑帅 张凯 高雨秋 于雪 ZHENG Shuai;ZHANG Kai;GAO Yu-Qiu;YU Xue(Department of Pharmacy,Hongqi Hospital Affiliated to Mudanjiang Medical College,Mudanjiang 157000,China)

机构地区：[1]牡丹江医学院附属红旗医院药学部,黑龙江牡丹江157000

出　　处：《中国病理生理杂志》2021年第12期2197-2204,共8页Chinese Journal of Pathophysiology

基　　金：黑龙江省卫生健康委科研课题(No.2020-410)。

摘　　要：目的:探究苦参总黄酮(TF-SF)是否通过调控核因子E2相关因子2/血红素加氧酶1(Nrf2/HO-1)通路影响大鼠肝纤维化(HF)。方法:将SD大鼠随机分为对照组、HF组、低剂量苦参总黄酮组、中剂量苦参总黄酮组、高剂量苦参总黄酮组及高剂量苦参总黄酮+全反式维甲酸(Nrf2抑制剂)组,每组10只;四氯化碳-花生油溶液灌胃饲养8周构建HF大鼠模型,于第9周开始低、中、高剂量苦参总黄酮组及高剂量苦参总黄酮+全反式维甲酸组大鼠分别给予相应等剂量药物进行灌胃,而对照组和HF组大鼠予以等体积生理盐水灌胃(每天1次),为期4周,实验结束后观察各组大鼠一般情况。ELISA法检测血清超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、透明质酸(HA)、Ⅳ型胶原(Ⅳ-C)、层黏连蛋白(LN)和Ⅲ型前胶原肽(PⅢP)水平;HE及Masson染色观察大鼠肝脏组织病理学情况;Western blot检测大鼠肝脏组织中Nrf2/HO-1通路蛋白表达水平;real-time PCR检测肝脏组织中Nrf2和HO-1 mRNA表达水平。结果:与对照组相比,HF组大鼠活动迟钝、体态瘦弱、食欲不振,肝脏质地发硬、边缘钝化、颜色暗淡且被膜缺乏;与HF组相比,低、中、高剂量苦参总黄酮组大鼠活动、饮食及肝脏形态均得到改善;与高剂量苦参总黄酮组相比,高剂量苦参总黄酮+全反式维甲酸组大鼠活动、饮食及肝脏不良状况加重。与对照组相比,HF组大鼠饮食量,饮水量,体重,血清SOD和GSH-Px水平,以及肝脏组织Nrf2和HO-1蛋白及mRNA水平均显著降低,而肝脏重量,血清MDA、AST、ALT、HA、Ⅳ-C、LN和PⅢP水平,以及HF分期和半定量评分均显著增加(P<0.05);与HF组相比,低、中、高剂量苦参总黄酮组大鼠饮食量,饮水量,体重,血清SOD和GSH-Px水平,以及肝脏组织Nrf2和HO-1蛋白及mRNA水平均显著增加,而肝脏重量,血清MDA、AST、ALT、HA�AIM:To explore whether total flavonoids of Sophorae flavescens(TF-SF)affects hepatic fibrosis(HF)in rats by regulating the nuclear factor E2-related factor 2/heme oxygenase-1(Nrf2/HO-1)signaling pathway.METHODS:The SD rats were randomly divided into control group,HF group,low-dose TF-SF group,middle-dose TFSF group and high-dose TF-SF group and high-dose TF-SF+all-trans retinoic acid(ATRA;Nrf2 inhibitor)group,with 10 rats in each group.The HF rat model was constructed by feeding with carbon tetrachloride(CCl4)-peanut oil solution for 8 weeks,starting from the 9 th week.The rats in low-,middle-and high-dose TF-SF groups and high-dose TF-SF+ATRA group were given corresponding doses of drugs by gavage,while the rats in control group and HF group were given the same volume of normal saline intragastrically,once per day for 4 weeks.The general conditions of the rats in each group were observed after the experiment.ELISA method was used to measure the levels of serum superoxide dismutase(SOD),malondialdehyde(MDA),glutathione peroxidase(GSH-Px),aspartate aminotransferase(AST),alanine aminotransferase(ALT),hyaluronic acid(HA),type IV collagen(IV-C),laminin(LN)and procollagen typeⅢpeptide(PⅢP).HE staining and Masson staining were used to observe the pathological changes of rat liver tissues.The protein levels of Nrf2/HO-1 signaling pathway-related molecules in rat liver tissues were determined by Western blot.The mRNA expression of Nrf2 and HO-1 in liver tissues was detected by real-time PCR.RESULTS:Compared with control group,the rats in HF group were sluggish in activity,weak in posture and loss of appetite,the liver texture was hard,the edges were dull,the color was dull and the capsule was lacking.Compared with HF group,the activity,diet and liver morphological changes of the rats in low-,middle-and high-dose TF-SF groups were improved.Compared with high-dose TF-SF group,the activity,diet and liver dysfunction of the rats in high-dose TF-SF+ATRA group were worsened.Compared with control group,food intake,water int

关 键 词：苦参总黄酮 肝纤维化 核因子E2相关因子2 血红素加氧酶1 氧化应激 

分 类 号：R575.2[医药卫生—消化系统] R363[医药卫生—内科学]