不同途径移植大鼠骨髓间充质干细胞对大鼠肝硬化的作用研究

Effect of Different Transplantation Methods of Bone Marrow Mesenchymal Stem Cells on Rats with Liver Cirrhosis

目的探索不同途径移植绿色荧光蛋白(green fluorescent protein,GFP)标记的大鼠骨髓间充质干细胞(rat bone marrow mesenchymal stem cells,rBMSCs)在慢性肝纤维化中的作用,以及移植后细胞在肝脏组织内的存活、分布情况。方法全骨髓差速贴壁法分离、培养BMSCs,利用携带GFP基因的慢病毒载体感染P3代BMSCs,建立四氯化碳慢性肝纤维化大鼠模型,通过不同途径移植GFP标记的BMSCs,移植后维持四氯化碳肝损伤,1月后处死大鼠,行肝脏功能相关指标白蛋白(albumin,ALB)、谷丙转氨酶(alaninotransferase,ALT)和谷草转氨酶(aspartate aminotrans-ferase,AST)检测,观察肝脏局部组织标本苏木精-伊红染色(hematoxylin-eosin staining,HE)、胶原纤维染色和荧光分布情况。结果不同途径BMSCs移植对大鼠四氯化碳所致慢性肝纤维化肝脏功能无明显改善,移植后GFP标记的BM-SCs均可向损伤肝脏迁移或停留,不同途径移植的BMSCs在肝脏内分布不同。结论大鼠骨髓间充质干细胞移植对大鼠慢性肝纤维化无明显治疗作用,可能参与了肝纤维化的形成。

Objective To investigate the effects of transplantation of rat bone marrow mesenchymal stem cells（rBMSCs） through different injection ways on the development of fibrogenesis in rats,and observe the distribution of transplanted BMSCs in the injured liver.Methods BMSCs were isolated and purified by differential adhention methods.The third passage of cells were identified and labeled by green fluorescent protein（GFP） gene transfection.Experimental animals were randomly divided into control and hepatocirrhosis groups.The hepatocirrhosis groups received CCl4 by gavage twice a week for 8 weeks.The GFP labeled-BMSCs were injected into the rats with liver fibrosis by 4 common transplantation ways.All rats were sacrificed 4 weeks after injection and their hepatic function were examined.The degree of liver cirrhosis and the distribution of the infused cells were observed using the liver tissue slices.Results These transplanted BMSCs could survive,migrate to the injured liver,and differently distributed according to the injected ways.The levels of plasma albumin（ALB）,alaninotransferase（ALT） and aspartate aminotransferase（AST） were all decreased and the liver fibrosis was further developed in the BMSCs transplanted groups.Conclusion BMSCs transplantation has no therapeutic effects on chronic liver fibrosis of rats,but may participate in the cirrhosis development.