胆管结扎诱导的梗阻性胆汁淤积对大鼠肝脏肝细胞影响的初步探究

doi:10.20223/j.cnki.1000-8535.2025.09.002

广州医药 ›› 2025, Vol. 56 ›› Issue (9): 1165-1174.DOI: 10.20223/j.cnki.1000-8535.2025.09.002

胆管结扎诱导的梗阻性胆汁淤积对大鼠肝脏肝细胞影响的初步探究

黄宇^1,2, 宫本大辅², 李天洋^1,2, 李佩霖², 翁杰锋¹, 古维立³

1 华南理工大学附属第二医院（广州市第一人民医院）肝胆胰外科（广东广州 510180）;
2 日本长崎大学大学院移植・消化器外科（日本长崎 8528501）;
3 广州医科大学附属中医医院肝胆外科（广东广州 510640）

收稿日期:2024-12-01 出版日期:2025-09-20 发布日期:2025-10-31
作者简介:黄宇华南理工大学附属第二医院（广州市第一人民医院）肝胆胰外科主治医师,广州医科大学外科学硕士,日本长崎大学医学博士; 现为广东省基层医药学会转移性肝癌专业委员会委员,广州市医师协会青年医师分会委员会委员,广州市医师协会肝胆胰外科医师分会委员会委员,广州市医学会加速康复外科学分会委员会委员; 任《广州医药》第一届青年编委,《中国普通外科杂志》第九届中青年编委,eGastroenterology第一届青年通讯编委; 致力于肝胆胰外科疾病的临床诊疗和科研工作,洞悉本专业领域国内外研究动向,尤其擅长荧光辅助下胆囊胆道等良性疾病的微创外科治疗; 主持广东省医学科研基金项目课题1项,广州市科技计划项目课题2项,广州市中医药和中西医结合科技项目课题1项; 获得实用新型技术专利2项; 在国内期刊杂志上发表论文20余篇,以第一或通信作者发表 SCI 论文10篇
基金资助:
广东省医学科研基金项目（A2024088）; 广州市科技计划项目（2024A03J1016）; 广州市中医药和中西医结合科技项目（20242A011001）

Preliminary study of bile duct ligation induced obstructive cholestasis in rat hepatocytes

HUANG Yu^1,2, MIYAMOTO Daisuke², LI Tianyang^1,2, LI Peilin², WENG Jiefeng¹, GU Weili³

1 Department of HBP Surgery, Guangzhou First People’s Hospital, the Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou 510180, China;
2 Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki 8528501, Japan;
3 Department of Hepatobiliary Surgery, the Affiliated TCM hospital of Guangzhou Medical University, Guangzhou 510640, China

Received:2024-12-01 Online:2025-09-20 Published:2025-10-31

摘要/Abstract

摘要： 目的初步探究胆管结扎诱导的梗阻性胆汁淤积对大鼠肝细胞的影响。方法 10只Lewis大鼠随机分为对照组和胆汁淤积组,每组各5只,胆汁淤积组采用胆管结扎2周诱导梗阻性胆汁淤积大鼠模型。苏木精-伊红染色和苯胺蓝染色比较组织病理变化,使用生化分析比较两组小鼠肝功能情况。采用改良的两步胶原酶灌注分离原代肝细胞,通过RT-qPCR检测两组小鼠肝细胞标志基因、细胞增殖标志基因以及胆管细胞标志基因的表达情况。结果与对照组相比,胆汁淤积组肝脏表现为明显的肝组织紊乱和纤维胶原蛋白沉积以及肝功能的损害。胆汁淤积组较对照组的原代肝细胞更高表达细胞增殖标志基因：细胞增殖标志物（Ki67）基因,叉头盒M1蛋白（Foxm1）基因,增殖细胞核抗原（Pcna）基因和肝细胞生长因子（HGF）基因（P<0.05）;胆汁淤积组的原代肝细胞表达更低水平的肝细胞标志基因：白蛋白（Alb）基因,多药耐药相关蛋白2（Mrp2）基因,胆盐输出泵（Bsep）基因和肝细胞连环蛋白1（Catenin1）基因（P<0.05）,同时表达更高水平的胆管细胞标志基因：细胞角蛋白7（Ck7）基因,细胞角蛋白 19（Ck19）基因,胆管细胞多药耐药性蛋白1（Mdr1）基因和胆管细胞囊性纤维化跨膜传导调节因子（Cftr）基因（P<0.05）以及肝祖细胞标志基因：上皮细胞黏附分子（Epcam）基因和Y染色体性别决定区-盒转录因子9（Sox9）基因（P<0.05）。结论胆汁淤积可诱导肝细胞向胆管细胞特性转化的可塑性。

关键词: 胆汁淤积性肝病, 肝细胞, 胆管细胞, 细胞可塑性, 胆管结扎

Abstract: Objective To explore the effect of bile duct ligation induced obstructive cholestasis on rat hepatocytes. Methods Ten Lewis rats were randomly divided into control group and cholestasis group,and the cholestasis was induced by bile duct ligation for 2 weeks. The histopathological changes were compared by H&E and aniline blue staining and the liver function was compared by biochemical analysis. Primary hepatocytes were isolated by modified two-step collagenase perfusion,and the expressions of hepatocyte marker genes,cell proliferation marker genes and cholangiocyte marker genes were detected by RT-qPCR. Results Compared with the control group,the liver of the cholestatic group showed obvious disordered histopathology,deposition of fibrous collagen and impaired liver function. Compared with the control group,the primary hepatocytes in the cholestasis group expressed higher cell proliferation-related genes（Ki67,Foxm1,Pcna and HGF）（P<0. 05）. Primary hepatocytes in the cholestasis group expressed lower levels of hepatocyte marker genes（Alb,Mrp2,Bsep and Catenin1）（P<0. 05）,and higher levels of cholangiocyte marker genes（Ck7,Ck19,Mdr1 and Cftr）（P<0. 05）and higher levels of the hepatic progenitor cell marker genes（Epcam and Sox9）（P<0. 05）. Conclusions Cholestasis induces rat hepatocyte plasticity in the transformation into bile duct properties.

Key words: cholestatic liver disease, hepatocyte, cholangiocyte, cell plasticity, bile duct ligation

黄宇, 宫本大辅, 李天洋, 李佩霖, 翁杰锋, 古维立. 胆管结扎诱导的梗阻性胆汁淤积对大鼠肝脏肝细胞影响的初步探究[J]. 广州医药, 2025, 56(9): 1165-1174.

HUANG Yu, MIYAMOTO Daisuke, LI Tianyang, LI Peilin, WENG Jiefeng, GU Weili. Preliminary study of bile duct ligation induced obstructive cholestasis in rat hepatocytes[J]. Guangzhou Medical Journal, 2025, 56(9): 1165-1174.

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胆管结扎诱导的梗阻性胆汁淤积对大鼠肝脏肝细胞影响的初步探究

Preliminary study of bile duct ligation induced obstructive cholestasis in rat hepatocytes

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