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  

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

作　　者：黄山 金礼权 喻宇 黄吉 张峻华 陈奕明 Huang Shan;Jin Liquan;Yu Yu;Huang Ji;Zhang Junhua;Chen Yiming(The First Affiliated Hospital of Dali University,Dali,Yunnan 671000,China;Institute of Translational Medicine for Metabolic Diseases,Dali University,Dali,Yunnan 671000,China)

机构地区：[1]大理大学第一附属医院,云南大理671000 [2]大理大学代谢性疾病转化医学研究院,云南大理671000

出　　处：《大理大学学报》2023年第4期22-27,F0003,共7页Journal of Dali University

基　　金：云南省教育厅科学研究基金项目(2020Y0571)。

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HOC在EGF、SCF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HOC中SOX9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HOC向BEC的分化过程。Objective:To investigate the expression changes of SOX9 during the directed differentiation of mouse hepatic oval cell(HOC)into biliary epithelial cell(BEC).Methods:A 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 centrifugation,and then induced to differentiate into BEC by using epidermal growth factor(EGF),stem cell growth factor(SCF)and leukemia inhibitory factor(LIF)in vitro.Cell morphological changes before and after differentiation were observed under inverted microscope,BEC were identified by flow cytometry,and the expression of SOX9 before and after differentiation was detected by immunofluorescence.Results:HOC were differentiated into purer BEC up to the 8th generation under the joint action of EGF,SCF and LIF.SOX9 was mainly expressed in the cytoplasm of primary mouse HOC,and gradually entered the nucleus during differentiation,with a significant increase in average light density value.Conclusion:SOX9 enters the nucleus and its expression remarkably increases during differentiation of mouse HOCs into BECs,indicating its possible involvement in the regulation of this differentiation process.

关 键 词：肝卵圆细胞 肝祖细胞 定向分化 胆管上皮细胞 SOX9 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]