损伤肝组织匀浆与骨髓间充质干细胞的分化

Effects of tissue homogenate from injured liver on induced differentiation of bone marrow mesenchymal stem cells

背景:目前认为,骨髓间充质干细胞能否分化为肝细胞与微环境关系密切。目的:探讨急性酒精性肝损伤组织匀浆对骨髓间充质干细胞的诱导分化作用。方法:分别用正常和损伤肝脏组织匀浆诱导骨髓间充质干细胞分化,在第0,3,7,10,14,21天用RT-PCR和Westernblot法检测甲胎蛋白、白蛋白的表达。结果与结论:骨髓间充质干细胞在正常及损伤的肝脏组织匀浆诱导下均表现为类上皮样改变。两组均从第3天开始检测到甲胎蛋白的表达,并在第7天左右达到高峰,之后表达逐渐减少,第14天时量很少;在第7天检测到白蛋白的表达,且随时间延长表达量逐渐增加,mRNA水平与蛋白水平表达一致。表明肝细胞在诱导培养下逐渐从幼稚到成熟。但正常和损伤肝组织匀浆诱导的肝细胞甲胎蛋白和白蛋白的表达量差异无显著性意义(P>0.05)。结果说明急性酒精性肝损伤组织匀浆可以诱导骨髓间充质干细胞向肝细胞分化,其诱导效果与正常肝脏组织匀浆相近。

BACKGROUND： Whether bone marrow mesenchymal stem cells can differentiate into hepatocytes is closely related to microenvironment. OBJECTIVE： To explore the effects of the acute alcoholic liver injury tissue homogenate on induced differentiation of bone marrow mesenchymal stem cells. METHODS： BMSCs were isolated from Sprague-Dawley rats, cultured, purified and identified in vitro. Rat models of alcohol-induced acute liver injury were prepared. Tissue homogenate of normal and injured liver was used to induce differentiation of bone marrow mesenchymal stem cells. The a-fetoprotein and albumin expression was detected at 0, 3 7, 10, 14, and 21 days using RT-PCR and western blot methods, respectively. RESULTS AND CONCLUSION： After induced by tissue homogenate of normal and injured liver, bone marrow mesenchymal stem cells exhibit epithelioid change. After induced by each tissue homogenate, a-fetoprotein expression was detected from the 3r~ day, peaked at about 7 days, decreased thereafter, and exhibited a very low level by 14 days; albumin expression was detected from the 7th day, and the expression level was gradually increased with time going. The mRNA and protein expression level was consistent. These findings demonstrate that hepatocytes gradually developed till mature. There was no significant difference in a-fetoprotein or albumin expression after induced by tissue homogenate of normal and injured liver （P 〉 0.05）. The liver tissue homogenates of acute alcoholic liver injury can induce bone marrow mesenchymal stem cells to differentiate into hepatocytes, with similar effects to normal liver tissue homogenate.