连翘提取物对对乙酰氨基酚诱导小鼠肝损伤的保护作用  

Protective Effect of Forsythiae suspensa Extract onAcetaminophen-induced Liver Injury in Mice

作　　者：赵晨栋 王萌[1] 张昊 代国年 安志霞 沈雅丽 王桂荣[1] ZHAO Chendong;WANG Meng;ZHANG Hao;DAI Guonian;AN Zhixia;SHEN Yali;WANG Guirong(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院,兰州730070

出　　处：《中国畜牧兽医》2021年第10期3845-3854,共10页China Animal Husbandry & Veterinary Medicine

基　　金：甘肃农业大学公招博士科研启动基金(GSAU-RCZX201702);大学生SRTP项目(202003048);兰州市人才创新创业项目(2019-RC-121)。

摘　　要：试验旨在探究连翘提取物(FSE)预防性干预对对乙酰氨基酚(acetaminophen,APAP)诱导小鼠肝损伤的保护作用及其潜在作用机制。采用半仿生-生物酶法提取连翘有效成分,使用APAP构建小鼠肝损伤模型。随机将60只雌性昆明小鼠分入正常对照(NC)组、APAP肝损伤模型(LD)组、连翘提取物(FSE)对照组、连翘提取物高剂量(HFSE+LD)组、连翘提取物中剂量(MFSE+LD)组和连翘提取物低剂量(LFSE+LD)组,每组10只。HFSE+LD组、MFSE+LD组、LFSE+LD组分别按每天200、100、50μg/g灌胃给予连翘提取物,NC组和LD组分别灌胃等量生理盐水,每天2次,连续给药6 d。预防性给药3 d后,腹腔注射APAP,每天1次。末次给药12 h后,试验小鼠眼球采血并快速取出肝脏,检测血清中丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)活性,以评价肝损伤程度;检测肝脏匀浆液中还原型谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、丙二醛(MDA)和过氧化氢(H2O2)水平,以评价肝脏氧化应激程度;制作肝脏病理切片,经苏木精伊红(HE)染色后观察肝脏病理变化;采用探针药物法测定肝脏线粒体细胞色素P4502E1(CYP2E1)活性;采用实时荧光定量PCR检测肝脏线粒体CYP2E1 mRNA表达水平;采用Western blotting法检测肝脏CYP2E1蛋白表达情况。结果表明:与NC组相比,LD组小鼠血清中ALT、AST活性均显著增加(P<0.05);肝脏SOD活性、GSH含量均显著降低(P<0.05),MDA、H2O2含量,CYP2E1 mRNA及蛋白表达水平均显著升高(P<0.05),成功构建小鼠肝损伤模型。200、100和50μg/g的连翘提取物可显著降低肝损伤小鼠血清ALT和AST活性(P<0.05),显著提高肝脏中SOD活性(P<0.05);200、100μg/g连翘提取物可显著提高肝脏中GSH水平(P<0.05),显著降低肝脏中MDA、H2O2水平及CYP2E1的mRNA和蛋白表达量(P<0.05)。以上结果表明,连翘提取物可减轻APAP诱导的小鼠肝损伤程度且呈剂量依赖性,其潜在作用机制可能与所含活性物质的抗氧�This study was aimed to explore the protective effect and potential mechanism of Forsythiae suspensa extract(FSE)preventive intervention on acetaminophen(APAP)-induced liver injury in mice.The semi-bionic enzyme method was used to extract the active components of Forsythia suspense,and the mouse liver injury model was constructed by APAP in this experiment.60 female Kunming mice were randomly divided into 6 groups:normal control(NC)group,APAP liver injury model(LD)group,Forsythia suspense extract(FSE)group,high-dose Forsythia suspense extract(HFSE+LD)group,medium-dose Forsythia suspense extract(MFSE+LD)group and low-dose Forsythia suspense extract(LFSE+LD)group.10 mice for each group.The HFSE+LD group,the MFSE+LD group,and the LFSE+LD group were intragastrically administrated with Forsythia suspense extract 200,100,and 50μg/g,respectively,twice a day,for 6 consecutive days.The NC group and the LD group were given the same amount of normal saline.After preventive administration with FSE for 3 days,APAP was injected intraperitoneally once a day.Blood samples were collected from the eyeball,and the liver was quickly taken out 12 h after the last administration.Serum aminotransferase(ALT)and aspartate aminotransferase(AST)were detected to evaluate the degree of liver injury.The homogenates of liver glutathione(GSH),superoxide dismutase(SOD),malondialdehyde(MDA)and hydrogen peroxide(H2O2)were detected to evaluate the degree of liver oxidative stress of liver.Liver samples were visualized by hematoxylin and eosin staining.Furthermore,liver mitochondrial cytochrome P4502E1(CYP2E1)activity were measured by probe drug method.The expression level of liver mitochondrial CYP2E1 mRNA was detected by Real-time quantitative PCR and the liver CYP2E1 protein expression was detected by Western blotting.The results showed that:Compared with NC group,the ALT and AST activities in serum were increased significantly(P<0.05),the SOD activity and GSH content in liver were significantly reduced(P<0.05),the MDA and H2O2 contents,and the

关 键 词：连翘提取物 对乙酰氨基酚 细胞色素P4502E1(CYP2E1) 肝损伤 

分 类 号：S853.74[农业科学—临床兽医学]