大理大学学报 ›› 2023, Vol. 8 ›› Issue (4): 22-27.

• 基础医学 • 上一篇    下一篇

SOX9在小鼠肝卵圆细胞定向分化为胆管上皮细胞过程中的表达

黄 山,金礼权,喻 宇,黄 吉,张峻华,陈奕明   

  1. 1. 大理大学第一附属医院,云南大理 6710002.大理大学代谢性疾病转化医学研究院,云南大理 671000

  • 收稿日期:2022-05-03 修回日期:2022-06-29 出版日期:2023-04-15 发布日期:2023-04-25
  • 通讯作者: 陈奕明,副教授,博士,E-mail:drchenyiming@gmail.com。
  • 作者简介:黄山,硕士研究生,主要从事肝胆外科疾病的临床诊治和基础研究。
  • 基金资助:

    云南省教育厅科学研究基金项目(2020Y0571

Expression of SOX9 in the Directed Differentiation of Mouse Hepatic Oval Cell into Biliary Epithelial Cells

Huang ShanJin LiquanYu Yu1Huang JiZhang JunhuaChen Yiming   

  1. 1. The First Affiliated Hospital of Dali UniversityDaliYunnan 671000China2. Institute of Translational Medicine for Metabolic DiseasesDali UniversityDaliYunnan 671000China

  • Received:2022-05-03 Revised:2022-06-29 Online:2023-04-15 Published:2023-04-25

摘要:

目的:研究转录因子SOX9在小鼠肝卵圆细胞(HOC)定向分化为胆管上皮细胞(BEC)过程中的表达变化。方法:通过 2-乙酰氨基芴灌胃联合2/3肝切除术构建小鼠HOC活化模型,利用二步酶消化法及Percoll 密度梯度离心法提取HOC后于体外条件下联合利用表皮生长因子(EGF)、干细胞生长因子(SCF)、白血病抑制因子(LIF)诱导HOC定向分化为BEC。于倒置显微镜下观察分化前后细胞形态变化,利用流式细胞术对BEC进行鉴定,免疫荧光法检测分化前后SOX9的表达。结果:小鼠原代HOCEGFSCF以及LIF联合作用下分化至第8代可获得较为纯化的BEC。在原代HOCSOX9主要表达于胞质,随着分化的进展逐渐表达于胞核,同时平均光密度值在分化过程中显著上调。结论:SOX9在小鼠HOC定向分化为BEC过程中由胞质进入胞核并且表达量显著升高,SOX9可能参与调控小鼠HOCBEC的分化过程。

关键词:

"> font-size:10.5pt, ">肝卵圆细胞, 肝祖细胞, 定向分化, 胆管上皮细胞, SOX9

Abstract:

ObjectiveTo investigate the expression changes of SOX9 during the directed differentiation of mouse hepatic oval cell HOCinto biliary epithelial cellBEC. MethodsA mouse HOC activation model was established by 2-acetylaminofluorene gavage and 2/3 partial hepatectomy. HOC were extracted by two-step enzyme digestion and Percoll density gradient centrifugationand then induced to differentiate into BEC by using epidermal growth factorEGF), stem cell growth factorSCFand leukemia inhibitory factorLIFin vitro. Cell morphological changes before and after differentiation were observed under inverted microscopeBEC were identified by flow cytometryand the expression of SOX9 before and after differentiation was detected by immunofluorescence.ResultsHOC were differentiated into purer BEC up to the 8th generation under the joint action of EGFSCF and LIF. SOX9 was mainly expressed in the cytoplasm of primary mouse HOCand gradually entered the nucleus during differentiationwith a significant increase in average light density value. ConclusionSOX9 enters the nucleus and its expression remarkably increases during differentiation of mouse HOCs into BECsindicating its possible involvement in the regulation of this differentiation process.

Key words:

"> font-size:10.5pt, ">hepatic oval cell, hepatic progenitor cell, directed differentiation, biliary epithelial cell, SOX9

中图分类号: