部分胆管结扎致胆汁淤积小鼠模型的构建方法学研究  

Method for constructing a mouse model of bile stasis caused by partial bile duct ligation

作　　者：屠海烨 包方奇 张利棕[3] 蒋琛 温思思 赵梓宇 方明笋[3] 陈民利[3,4] TU Haiye;BAO Fangq;ZHANG Lizong;JIANG Chen;WEN Sisi;ZHAO Ziyu;FANG Mingsun;CHEN Minl(School of Pharmacy,Zhejiang Chinese Medical University,Hangzhou 310053,China;Shanghai Medical College of Fudan University,Shanghai 200032,China;Animal Experimental Research Center,Zhejiang Chinese Medical University,Hangzhou 310053,China;Hangzhou Lifutai Biotechnology Co.,Ltd.,Hangzhou 310051,China;School of First Clinical,Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学药学院,杭州310053 [2]复旦大学上海医学院,上海200032 [3]浙江中医药大学动物实验研究中心,杭州310053 [4]杭州利孚泰生物科技有限公司,杭州310051 [5]浙江中医药大学第一临床医学院,杭州310053

出　　处：《中国实验动物学报》2024年第5期620-629,共10页Acta Laboratorium Animalis Scientia Sinica

基　　金：浙江省科技计划(2023C03004)。

摘　　要：目的 观察不同结扎位点和禁食方法对部分胆管结扎(partial bile duct ligation, pBDL)致胆汁淤积C57BL/6J小鼠模型的影响,研究成模率高、症状典型、稳定性好的pBDL造模方法。方法 分别采用选择性结扎左肝胆管(L-pBDL法)和左侧与正中胆管汇合处(ML-pBDL法)进行造模,观察不同pBDL结扎法对C57BL/6J小鼠血清谷丙转氨酶、谷草转氨酶、碱性磷酸酶(alkaline phosphatase, ALP)、总胆红素、总胆汁酸和肝组织病理学的影响。同时,通过术前分别禁食12 h和禁食16 h、术后均禁食禁水4 h,观察不同禁食方法对ML-pBDL模型体症和肝损伤的影响。结果 (1)ML-pBDL组黄疸出现率52.94%、术后3周内存活率64.71%,而L-pBDL组黄疸出现率11.76%、术后3周内存活率82.35%;与假手术组比较,L-pBDL组和ML-pBDL组血清肝功能指标均显著升高(P<0.01),且ML-pBDL组ALP活性明显高于L-pBDL组(P<0.05);相对于L-pBDL组,ML-pBDL组术后3周的肝纤维化更严重(P<0.01);(2)禁食16 h组黄疸出现率93.33%、术后3周内存活率73.77%,而禁食12 h组黄疸出现率42.86%、术后3周内存活率71.42%;与假手术组比较,禁食16 h组和禁食12 h组ALP活性、谷草转氨酶/谷草转氨酶比值、总胆汁酸水平与胶原纤维面积占比均显著升高(P<0.05)。禁食16 h组各观察指标均高于禁食12 h组,但无显著性差异(P>0.05),禁食12 h和禁食16 h组均出现明显的胆管增生与肝纤维化(P<0.01),且禁食16 h组纤维化更严重(P<0.01)。结论 L-pBDL和ML-pBDL结扎法均可建立小鼠胆汁淤积模型,但L-pBDL模型症状仅表现为一过性的损伤特征,而ML-pBDL模型的肝病变典型、症状稳定,且适当延长术前禁食时间可提高ML-pBDL模型的成模率和稳定性,且病理症状更典型。Objective To observe the effects of different ligation sites and fasting method on a C57BL/6J mouse model of partial bile duct ligation(pBDL)-induced cholestasis,to establish a pBDL modeling method with a high modeling rate,typical symptoms,and good stability.Methods C57BL/6J mice were subjected to selective ligation of the left hepatic bile duct(L-pBDL)and left-to-median bile duct junction ligation(ML-pBDL)for modeling,and the effects of different pBDL ligation method on serum alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase(ALP),total bilirubin,total bile acid,and liver histopathology were observed.The effects of different fasting method on symptoms and liver injury in the ML-pBDL model were also observed after fasting for 12 and 16 h before surgery,and for 4 h after surgery.Results(1)The incidence of jaundice in the ML-pBDL group was 52.94%and the survival rate within 3 weeks after surgery was 64.71%,while the incidence of jaundice in the L-pBDL group was 11.76%and the survival rate within 3 weeks after surgery was 82.35%.Compared with those in the sham surgery group,serum liver function indicators were significantly increased in the L-pBDL and ML-pBDL groups(P<0.01),and ALP activity was significantly higher in the ML-pBDL group than in the L-pBDL group(P<0.05).Compared with mice in the L-pBDL group,mice in the MLpBDL group had more severe liver fibrosis at 3 weeks post-surgery(P<0.01).(2)In addition,the incidence of jaundice in the 16 h fasting group was 93.33%and the survival rate within 3 weeks after surgery was 73.77%,while the incidence of jaundice in the 12 h fasting group was 42.86%and the survival rate within 3 weeks after surgery was 71.42%.Compared with those in the normal group,ALP activity,alanine aminotransferase/aspartate aminotransferase ratio,total bile acid level,and proportion of collagen fiber area were all significantly increased in the 16 h and 12 h fasting groups(P<0.05).Although the observed indicators were higher in the 16 h fasting group compared with those in t

关 键 词：C57BL/6J小鼠 部分胆管结扎 胆汁淤积模型 肝损伤 

分 类 号：Q95-33[生物学—动物学]