金丝桃苷改善吡咯里西啶生物碱诱导小鼠急性肝损伤的作用及机制研究  

Effect and mechanism of hyperoside on acute liver injury induced by pyrrolizidine alkaloids in mice

作　　者：徐婕 陈依琳 阮德清 张雨萌 王汛江 丁丽丽[1,2] 王峥涛 熊爱珍[1,2] 杨莉 XU Jie;CHEN Yilin;RUAN Deqing;ZHANG Yumeng;WANG Xunjiang;DING Lili;WANG Zhengtao;XIONG Aizhen;YANG Li(Chinese Materia Medica Institute of Shanghai University of Traditional Chinese Medicine,the MOE Key Laboratory for Standardization of Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines,Shanghai 201203,China;Shanghai R&D Center for Standardization of Traditional Chinese Medicines,Shanghai 201203,China)

机构地区：[1]上海中医药大学中药研究所,中药标准化教育部重点实验室暨国家中医药管理局中药新资源与质量评价重点实验室,上海201203 [2]上海中药标准化研究中心,上海201203

出　　处：《上海中医药杂志》2023年第12期80-87,共8页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82130115);上海市卫健委上海市进一步加快中医药事业发展三年行动计划项目(ZY[2021-2023]-0215)。

摘　　要：目的明确金丝桃苷对吡咯里西啶生物碱(PAs)诱导小鼠急性肝损伤的作用及机制。方法40只C57BL/6雄性小鼠随机分成5组,即空白对照组、模型组以及金丝桃苷低、中、高剂量组,每组8只。除空白对照组外,其余小鼠采用PAs建立急性肝损伤模型,造模6 h、30 h后金丝桃苷低、中、高剂量组小鼠分别灌胃给予20、40、80 mg/kg金丝桃苷各1次,造模48 h后,收集血清、肝脏、回肠、粪便。检测小鼠血清生化指标丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和总胆汁酸(TBA)水平;苏木精-伊红(HE)染色法观察小鼠肝脏组织病理学形态变化;测定小鼠血清、肝脏、回肠、粪便中胆汁酸含量;实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法及Western blot法检测小鼠肝脏组织中法尼酯X受体(Fxr)、小异源二聚体伴侣(Shp)、胆盐输出泵(Bsep)、胆固醇-7α-羟化酶(Cyp7a1)、甾醇-12α-羟化酶(Cyp8b1)mRNA与蛋白表达水平。结果与空白对照组比较,模型组血清生化指标ALT、AST、TBA水平显著升高(P<0.05),肝细胞大面积坏死,同时伴有肝窦淤血;与模型组比较,金丝桃苷各剂量组血清生化指标ALT、AST、TBA水平显著降低(P<0.05),金丝桃苷中、高剂量组未见明显肝细胞坏死及肝窦淤血。与空白对照组比较,模型组胆汁酸代谢异常,小鼠肝脏Fxr、Shp、Bsep、Cyp7a1、Cyp8b1 mRNA及蛋白表达水平明显降低(P<0.05);与模型组比较,金丝桃苷中剂量组胆汁酸稳态失衡有所改善,小鼠肝脏Fxr、Shp、Bsep、Cyp7a1、Cyp8b1 mRNA及蛋白表达水平明显升高(P<0.05)。结论金丝桃苷可改善PAs诱导的急性肝损伤,其机制与调控胆汁酸相关基因、维护胆汁酸稳态平衡有关。Objective To determine the effect and mechanism of hyperoside on acute liver injury induced by pyrrolizidine alkaloids(PAs)in mice.Methods Forty C57BL/6 male mice were randomly divided into 5 groups:blank control group,model group and low,medium and high dose hyperoside groups with 8 mice in each group.Except for the blank control group,the other mice were treated with PAs to establish the model of acute liver injury,and 6 h and 30 h later after the establishment of the model,the mice in the low,medium and high dose groups were given 20,40 and 80 mg/kg hyperoside respectively.After 48 hours,the serum,liver,ileum and feces were collected.The biochemical indexes of mice serum alanine aminotransferase(ALT),aspartate aminotransferase(AST)and total bile acid(TBA)were measured.The histopathological changes of liver were observed by hematoxylineosin(HE)staining.The contents of bile acid in serum,liver,ileum and feces of mice were determined.The expressions of farnesyl X receptor(Fxr),small heterodimer chaperone(Shp),bile salt output pump(Bsep),cholesterol-7α-hydroxylase(Cyp7a1),sterol-12α-hydroxylase(Cyp8b1)mRNA and protein in mouse liver were detected by real-time fluorescence quantitative reverse transcriptase polymerase chain reaction(RT-qPCR)and Western blot.Results Compared with the blank control group,the serum biochemical index levels of ALT,AST and TBA in the model group were significantly increased(P<0.05),and the large area necrosis of hepatocytes accompanied with hepatic sinusoid congestion was found in the model group,while the serum biochemical index levels of ALT,AST and TBA in all hyperoside groups were significantly lower than those in the model group(P<0.05),but no obvious hepatocyte necrosis and hepatic sinusoid congestion were found in the medium and high dose hyperoside groups.Compared with the blank control group,the bile acid metabolism in the model group was abnormal,and the expression levels of Fxr,Shp,Bsep,Cyp7a1,Cyp8b1 mRNA and protein in the liver of the model group were significantly decrea

关 键 词：急性肝损伤 吡咯里西啶生物碱 金丝桃苷 胆汁酸 小鼠模型 中药研究 

分 类 号：R285.5[医药卫生—中药学]