上调hHGF促进人骨髓间充质干细胞向肝样细胞分化  

Human hepatocyte growth factor promotes human bone marrow stem cells to differentiate into hepatocyte-like cells

作　　者：陈丽[1,2] 崔洁[1] 侯军霞[1] 贺莉[1] 郑淑梅[1] 曾维政[1] 蒋明德[1] 

机构地区：[1]成都军区总医院消化内科,成都610083 [2]第三军医大学研究生管理大队,重庆400038

出　　处：《免疫学杂志》2011年第1期37-41,48,共6页Immunological Journal

摘　　要：目的探讨人促肝细胞生长因子(hHGF)对人骨髓间充质干细胞(hBMSCs)向肝样细胞分化的影响。方法克隆hHGF基因,构建携带hHGF基因的真核表达质粒(pcDNA3.1(+)-hHGF),用脂质体法将pcDNA3.1(+)-hHGF转染hBMSCs,分别通过激光共聚焦、反转录聚合酶链反应(RT-PCR)、免疫蛋白印迹(Western blot)检测hBMSCs中表达hHGF的水平;用酒精性肝硬化患者血清诱导hHGF上调后的hBMSCs,采用Western blot分别检测诱导细胞内甲胎蛋白(AFP)及白蛋白(ALB)的表达变化。结果成功构建pcDNA3.1(+)-hHGF表达质粒;激光共聚焦、RT-PCR、Western blot结果均表明转染后hBMSCs中hHGF分子呈高表达,Western blot结果发现诱导7 d,14 d转染后hBMSCs中AFP、ALB表达明显高于对照组。结论转染pcDNA3.1(+)-hHGF可以明显上调hHGF在hBMSCs中的表达;hBMSCs高表达hHGF后,经酒精性肝硬化血清诱导可明显促进其向肝样细胞分化,为基因治疗终末期肝病提供了一种新方法。This study aimed to investigate the effects of human hepatocyte growth factor（hHGF） gene on the human bone marrow stem cells（hBMSCs） developing into hepatocyte-like cells.hHGF gene was inserted into pcDNA3.1＋,and then transfected into hBMSCs using Lipofectamine 2000.The expression of hHGF was confirmed to be up-regulated in the pcDNA3.1＋-HGF transfected hBMSCs by using confocal laser,RT-PCR and Western blot respectively.Then the pcDNA3.1＋-HGF transfected hBMSCs were cultured with the serum of alcoholic liver cirrhosis patients for 7 and 14 days.Western blot was performed to detect the alteration of α-fetoprotein（AFP） and albumin（ALB） expressions.At last,the recombinant plasmid pcDNA3.1＋-HGF was constructed successfully,and could effectively increase both the hHGF mRNA and protein expressions in hBMSCs.The expression of AFP and ALB obviously increased in the pcDNA3.1＋-HGF transfected hBMSCs after co-cultured with the serum of alcoholic liver cirrhosis patients for 7 and 14 days.We conclude that pcDNA3.1＋-hHGF transfected hBMSCs could apparently up-regulate the expression of hHGF.Furthermore exogenous hHGF gene could promote hBMSCs to differentiate into hepatocyte-like cells,which may be a promising gene therapeutic method for terminal stage hepatopaths.

关 键 词：人骨髓间充质干细胞 人促肝细胞生长因子 肝样细胞 甲胎蛋白 白蛋白 

分 类 号：R392.12[医药卫生—免疫学]