17β⁃雌二醇对rBMSCs肝祖细胞定向分化及Wnt信号通路的调控作用  

Regulation of 17β-estradiol on the differentiation of hepatic progenitor cells and Wnt signaling pathway in rBMSCs

作　　者：曾令锋 黄俊宇 熊瑶 张志东[1] 王景浩 黄硕[3] 韦宏成[2] ZENG Lingfeng;HUANG Junyu;XIONG Yao;ZHANG Zhidong;WANG Jinghao;HUANG Shuo;WEI Hongcheng(Department of pharmacy,The first affiliated hospital of Jinan University,Guangzhou,Guangdong,Chi⁃na,510630;Department of gastroenterology,The First Affiliated Hospital of Jinan University,Guang⁃zhou,Guangdong,China,510630;Departments of Radiology,Xiehe Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei,China,430030;The fifth affiliated hos⁃pital,Guangzhou medical university,Guangzhou,Guangdong,China,510700)

机构地区：[1]暨南大学附属第一医院药学部,广东广州510630 [2]暨南大学附属第一医院消化科人工肝,广东广州510630 [3]华中科技大学同济医学院附属协和医院放射科,湖北武汉430030 [4]广州医科大学附属第五医院,广东广州510700

出　　处：《分子诊断与治疗杂志》2020年第4期459-463,482,共6页Journal of Molecular Diagnostics and Therapy

基　　金：中央高校青年基金项目(11619347)。

摘　　要：目的探究17β⁃雌二醇(17β⁃E2)对大鼠骨髓间充质干细胞(rBMSCs)肝祖细胞定向分化能力及Wnt信号通路的调控作用。方法通过全骨髓贴壁法获得稳定的rBMSCs,将rBMSCs分为α⁃MEM培养基(Control)、肝向诱导培养基(经典)、17β⁃E2+α⁃MEM培养基(E2)、17β⁃E2+肝向诱导培养基(经典+E2)组,共4组,显微镜下观察形态学变化,发光免疫法检测细胞裂解液AFP含量,运用Western bolt法检测Wntβ⁃catenin信号蛋白(Wnt3a、β⁃catenin及LRP⁃5)表达。结果在形态学上,经典与经典+E2组的rBMSCs逐渐呈肝祖细胞样变化,而Control组和E2组无明显改变。经典组、经典+E2组AFP值显著高于Control组、E2组(P<0.05);经典+E2组AFP值显著高于经典组(P<0.05)。与Control组及E2组相比,经典组与经典+E2组β⁃Catenin、Wnt3a表达量显著降低(P<0.05),且经典+E2组β⁃Catenin、Wnt3a表达量显著低于经典组(P<0.05);而LRP⁃5的表达差异均无统计学意义(P>0.05)。结论17β⁃E2可促rBMSCs肝祖细胞向分化,其机制可能与调控Wntβ⁃catenin信号通路相关。Objective To investigate the regulatory effect of 17β⁃estradiol(17β⁃E2)on the differ⁃entiation of hepatic progenitor cells of rat bone marrow mesenchymal stem cells(rBMSCs)and Wnt signaling pathway.Methods rBMSCs were obtained by whole bone marrow adherence method.The cells were divided into four groups:α⁃MEM medium(control),liver⁃inducing medium(classical),and E2+α.⁃MEM medium(E2),E2+hepatic inducing medium(classical+E2),observed in morphology.The AFP concentration was measured using the luminescence immunoassay.The expression of Wnt signaling proteins,included Wnt3a,β-catenin and LRP⁃5,was detected by Western blot.Results Morphologically,rBMSCs in the classic and classic+E2 groups gradually showed liver progenitor⁃like changes,while the control group and E2 group showed no significant changes.The AFP concentration of classic group and classic+E2 group was significant⁃ly higher than that of the control group and E2 group(P<0.05).The AFP value of classic+E2 group was sig⁃nificantly higher than that of the classic group(P<0.05).Compared with the control group and the E2 group,the expression levels ofβ⁃Catenin and Wnt3a in the classic group and the classic+E2 group were significantly reduced(P<0.05),and the expression levels ofβ⁃Catenin and Wnt3a in the classic+E2 group were signifi⁃cantly lower than that in the classic group(P<0.05).There was no statistically significant difference in the ex⁃pression of LRP⁃5(P>0.05).Conclusion 17β⁃E2 can promote the differentiation of rBMSCs hepatic pro⁃genitor cells,and the mechanism may be related to the regulation of Wntβ⁃catenin signaling pathway.

关 键 词：骨髓间充质干细胞 17β⁃雌二醇 WNT信号通路 肝祖细胞向分化 

分 类 号：R575[医药卫生—消化系统]