中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (23): 3705-3709.doi: 10.3969/j.issn.2095-4344.1317

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

肌成纤维细胞在小鼠肝内胆管发育过程中的作用

杨均均1，李德卫2，王  伟2，冯  渊1   

  1.  (1川北医学院第二临床学院•南充市中心医院感染科，四川省南充市  637000；2重庆医科大学附属第一医院肝胆外科，重庆市   400016)
  • 收稿日期:2019-03-08 出版日期:2019-08-18 发布日期:2019-08-18
  • 通讯作者: 李德卫，博士，教授，重庆医科大学附属第一医院肝胆外科，重庆市 400016
  • 作者简介:杨均均，女，1985年生，湖南省绥宁县人，苗族，2012年重庆医科大学毕业，硕士，主治医师，主要从事肝病及感染性疾病方面的研究。
  • 基金资助:

    国家自然科学基金面上项目(81470898)，项目负责人：李德卫

Role of myofibroblasts in the development of mouse intrahepatic bile ducts 

Yang Junjun1, Li Dewei2, Wang Wei2, Feng Yuan1   

  1. (1Department of Infections, the Second Clinical College of North Sichuan Medical College, Nanchong Central Hospital, Nanchong 637000, Sichuan Province, China; 2Department of Hepatobiliary Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China)
  • Received:2019-03-08 Online:2019-08-18 Published:2019-08-18
  • Contact: Li Dewei, MD, Professor, Department of Hepatobiliary Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
  • About author:Yang Junjun, Master, Attending physician, Department of Infections, the Second Clinical College of North Sichuan Medical College, Nanchong Central Hospital, Nanchong 637000, Sichuan Province, China
  • Supported by:

    the National Natural Science Foundation of China (General Program), No. 81470898 (to LDW)

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摘要：

文章快速阅读：

文题释义：
细胞角蛋白19(cytokeratin19，CK19)：通常作为胆管细胞的标志，在人类肝祖细胞中低表达，但在鼠类肝祖细胞中不表达，在胆管细胞分化早期检测细胞角蛋白19存在技术上的困难。
肌成纤维细胞：是指含有肌动蛋白、肌球蛋白和其他肌肉蛋白的成纤维样细胞。收缩蛋白质排列成可具有收缩功能的形式。成纤维细胞及其表达的细胞因子是肿瘤组织重要的微环境成分，在促进肿瘤的增殖、转移和新生血管形成方面起着非常重要的作用。
摘要
背景：研究表明肌成纤维细胞具有促进肝祖细胞分化为胆管细胞作用，但对于肌成纤维细胞在体内胆管发育中作用仍不清楚。
目的：观察小鼠肝内胆管发育的特点，以及肌成纤维细胞在胆管发育过程中的作用。
方法：SPF级C57小鼠购自重庆医科大学实验动物中心，实验方案经重庆医科大学动物实验伦理委员会批准，批准号为2017-182。采用14.5 d、16.5 d孕鼠及刚出生(0 d)和出生后10 d的小鼠，麻醉后取其胎鼠肝脏(E14.5、E16.5)和出生后小鼠肝脏(P0、P10)，采用免疫组织化学技术观察α平滑肌肌动蛋白、Sox9及细胞角蛋白19在E14.5、E16.5小鼠胎肝和P0、P10小鼠肝脏中的表达。
结果与结论：①E14.5小鼠胎肝中，α平滑肌肌动蛋白阳性肌成纤维细胞环绕门静脉周围，Sox9阳性胆管细胞紧靠α平滑肌肌动蛋白阳性肌成纤维细胞，并形成单层细胞排列的胆管板；②E16.5小鼠胎肝中，紧靠α平滑肌肌动蛋白阳性肌成纤维细胞的细胞角蛋白19阳性胆管细胞形成双层胆管细胞结构；③P0小鼠肝脏中，细胞角蛋白19阳性胆管细胞发育形成具有管腔的胆管结构，并且α平滑肌肌动蛋白阳性肌成纤维细胞开始包绕胆管；④P10小鼠肝脏中，胆管进一步发育扩张，门静脉周围未形成胆管的胆管细胞开始消失。并且α平滑肌肌动蛋白阳性肌成纤维细胞环绕胆管，胆管旁发育形成肝动脉；⑤结果表明了小鼠肝内胆管发育的特点，发现α平滑肌肌动蛋白阳性肌成纤维细胞与肝内胆管发育的关系密切，且在肝内胆管发育中发挥了重要作用。

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程
ORCID: 0000-0002-5282-0129(杨均均)

关键词: 肌成纤维细胞, 胆管, 发育, 肝脏, 小鼠, 细胞角蛋白19, 国家自然科学基金

Abstract:

BACKGROUND: Existing evidence has shown that myofibroblasts can promote hepatic progenitor cells to differentiate into bile duct cells. But, the role of myofibroblasts in the development of bile duct in vivo remains unclear.
OBJECTIVE: To investigate the developmental features of intrahepatic bile duct in mice and role of myofibroblasts in the development of bile duct.
METHODS: C57 mice, SPF grade were proved by the Laboratory Animal Center of Chongqing Medical University, and the study was approved by the Ethics Committee of Experimental Animal Ethics of Chongqing Medical University, approval number: 2017-182. The mice in gestational day 14.5 and 16.5, postnatal 0 and 10 days (P0, P10) mice were selected. After anesthesia, the liver was removed from fetal mice of embryonic day 14.5 and 16.5 (E14.5, E16.5) and P0 and P10 mice. The expression levels of α smooth muscle actin, Sox9 and cytokeratin 19 in liver were examined by immunocytochemical method.
RESULTS AND CONCLUSION: (1) In the fetal liver of E14.5 mice, myofibroblasts positive for α smooth muscle actin were main located around the portal vein, which was adjacent to the bile duct cells positive for Sox9, and single layered ring called ductal plate formed. (2) In the fetal liver of E16.5 mice, the ductal plate positive for cytokeratin 19, which neared the myofibroblasts positive for α smooth muscle actin, became focally bilayered. (3) In the liver of P0 mice, the cholangiocytes positive for cytokeratin 19 formed bile ducts, which were surrounded by myofibroblasts positive for α smooth muscle actin. (4) In the liver of P10 mice, the bile duct expanded further, and cholangiocytes that could not form bile duct surrounding the portal vein disappeared. The bile duct was encircled by myofibroblasts positive for α smooth muscle actin, and a well-formed hepatic artery was found near the bile duct. (5) The developmental characteristics of intrahepatic bile ducts are systematic displayed. Myofibroblasts positive for α smooth muscle actin are closely related to intrahepatic bile duct development, and play critical role.

Key words: myofibroblasts, bile duct, development, liver, mice, cytokeratin 19, the National Natural Science Foundation of China

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