ANIT诱导亚急性肝内胆汁淤积大鼠模型肝脏氧化应激水平及胆汁酸受体表达的变化  

The changes of liver oxidative stress and bile acid receptor expression in subacute intrahepatic cholestasis rats induced by ANIT

作　　者：姚嘉明[1] 陈芝芸[2] 陈曦[3] 刘彬彬[1] 叶蔚[1] 王小奇[1] 张洁[1] Yao Jiaming

机构地区：[1]浙江中医药大学附属杭州市中医院,310007 [2]浙江中医药大学附属第一医院,310006 [3]四川省内江市第一人民医院,641000

出　　处：《浙江临床医学》2023年第1期15-17,21,共4页Zhejiang Clinical Medical Journal

基　　金：浙江省科技计划公益技术研究社会发展项目(2014C33238);浙江省中医药科技计划(2020ZB158)。

摘　　要：目的观察异硫氰萘酯(ANIT)诱导亚急性肝内胆汁淤积大鼠模型肝脏氧化应激水平及胆汁酸受体表达的变化。方法雄性SD大鼠随机分为正常组24只,模型组24只,模型组在第1、7、14、21天分别灌服1.6%ANIT80 mg/kg,正常组以等量麻油灌胃,分别在第9、16、23天各处理大鼠8只。各大鼠于处理前晚禁食不禁水,次日上午空腹麻醉下腹腔静脉取血,分离血清检测肝功能;分离肝组织,部分在10%中性甲醛中固定行病理观察,其余肝组织存入-80°C待用,黄嘌呤氧化酶法测定肝组织SOD活力,硫代巴比妥酸(TBA)法测定肝组织MDA含量,比色法测定肝组织GSH-PX和CAT活力,荧光定量PCR测定肝组织Nrf2、SOD1、SOD2、SOD3、CAT、FXR、Ntcp、Bsep、Mrp2、Mrp3基因的mRNA表达。结果模型组大鼠在每次给药后48 h血清ALT、AST、ALP、GGT、TBA、TBIL、DBIL水平比正常组明显升高(P<0.01),血清ALB水平比正常组明显下降(P<0.01),其在三个时相点均维持明显的肝内胆汁淤积,并出现早期肝纤维化表现。在三个时相点,模型组肝组织MDA含量均比正常组明显升高(P<0.01),而SOD、GSH-Px、CAT的活性比正常组明显降低(P<0.01),肝组织SOD1、SOD2、SOD3、CAT、FXR、Ntcp、Bsep、Mrp2的mRNA表达比正常组明显下降(P<0.05或P<0.01),Mrp3 mRNA比正常组明显增加(P<0.01);模型组肝组织Nrf2的mRNA表达在第16、23天比正常组明显下降(P<0.01)。结论ANIT间隙性多次给药可成功诱导大鼠亚急性肝内胆汁淤积,其发生过程中存在氧化应激及胆汁酸代谢障碍。Objective To observe the changes of liver oxidative stress and bile acid receptor expression in subacute intrahepatic cholestasis rats induced by Naphthalene isothiocyanate(ANIT).Methods Male SD rats were randomly divided into normal group and model group with 24 in each group.The rats in the model group were given 1.6%ANIT80 mg/kg on the 1st day,7th day,14th day and 21st day,and the rats in the normal group were given with equal capacity sesame oil.The rats were given fasting water on the night before the treatment,and their blood was collected from abdominal vein under fasting anesthesia on the morning of the next day to test liver function.The liver tissue was separated and part of the tissue was fixed in 10%neutral formaldehyde for pathology observation.The rest of the liver tissue was deposited with-80°C to be used.The xanthine oxidase method was used to determine liver tissue's SOD activity.The glucosinolates barbituric acid(associates)method was used to determine the liver tissue's MDA content.The colorimetric method was used to determine liver gsh-px and CAT activity.The fluorescence quantitative PCR method was used for the determination of liver tissue's Nrf2,SOD1,SOD2,SOD3,CAT,FXR,Ntcp,Bsep,Mrp2,Mrp3 gene mRNA expression.Results The model group rats in every 48 h after the treatment the serum ALT,AST,ALP,GGT,associates,TBIL,DBIL levels remained significantly higher than normal group(P<0.01).Compared with the normal group,the serum level of ALB was significantly decreased(P<0.01).All three phases lasted obvious intrahepatic cholestasis,and led to early liver fibrosis.At three time points,the MDA content in liver of model group was significantly higher than that of normal group(P<0.01),while SOD,GSH-Px and CAT activities were significantly lower than that of normal group(P<0.01).The mRNA expressions of SOD1,SOD2,SOD3,CAT,FXR,Ntcp,Bsep and Mrp2 in liver tissue were significantly decreased compared with normal group(P<0.05or P<0.01),while Mrp3 mRNA was significantly increased compared with normal group(P<0

关 键 词：肝内胆汁淤积 异硫氰酸萘酯 氧化应激 胆汁酸代谢 

分 类 号：R285.5[医药卫生—中药学]