骨髓间充质干细胞输注治疗大鼠小体积肝移植术后肝衰竭

Bone marrow mesenchymal stem cells infusion rescue the liver failure induced by small-for-size liver transplantation in rats

目的探讨大鼠骨髓间充质干细胞输注治疗大鼠小体积肝移植术后肝衰竭的可行性。方法体外分离、培养、鉴定大鼠骨髓间充质干细胞,采用细胞膜染料PKH26对骨髓间充质干细胞进行标记。将雄性SD大鼠随机分为对照组和治疗组。对照组于小体积（30%）肝移植术中,在移植肝血供恢复时自尾静脉注入磷酸盐缓冲液（PBS）0.5 ml,治疗组则在相应时段自尾静脉注入含5×105mmol/L骨髓间充质干细胞的PBS 0.5 ml。于术后1 d、2 d、3 d、5 d、7 d取血液及肝脏、脾脏标本,采用荧光显微镜观察标记的干细胞在肝脏及脾脏的分布变化。比较两组术后的ALT、AST、1周生存率,移植肝重量与原受体肝重量的比值。结果在术后早期,治疗组的肝脏中标记的骨髓间充质干细胞分布在大血管周围,此后逐渐进入肝实质中弥散分布;脾脏中标记的骨髓间充质干细胞随术后时间延长而显著减少。与对照组比较,治疗组术后1周生存率明显提高,术后3 d、5 d、7 d的ALT、AST明显降低,术后3 d、5 d移植肝重量与原受体肝重量的比值明显增高（P〈0.05~P〈0.01）。结论小体积肝移植后输注的骨髓间充质干细胞具有向移植肝脏定向迁移的能力,并可以对移植肝功能起到保护和支持作用。采用骨髓间充质干细胞治疗小体积肝移植术后肝衰竭具有可行性。

Objective To explore the feasibility of bone marrow mesenchymal stem cells（MSCs） infusion in rescuing the liver failure after the small-for-size liver transplantation in rats.Methods Rat MSCs were isolated,characterized,cultured,and then stained with cell membrane colourant PKH26.Male SD rats were randomly divided into two groups： control group and experiment group.In control group,0.5 ml phosphate buffer（PBS） were injected via tail vein in the small-for-size（30 %） liver transplantation.In experiment group,0.5 ml PBS containing 5×105 mmol/L MSCs was injected via tail vein.On day 1,2,3,5,7 after operation,the blood,liver and spleen samples were collected.Fluorescence microscope was used to observe the distribution of MSCs in liver and spleen.Alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,one week survival rate and the ratio between transplant and primary recipient liver weight were analyzed between the two groups after transplantation.Results In the early stage after transplantation,the labeled MSCs in liver were detected in experiment group,which were firstly located around the vessel restrictedly and then dispersive in liver parenchyma in the following days.The number of labeled MSCs in spleen was gradually decreased after transplantation.Compared with control group,one week survival rate after transplantation was significantly increased in experiment group.On day 3,5,7 after operation,ALT and AST in experiment group were much lower than that in control group.On day 3,5 after operation,the ratio between transplant and primary recipient liver weight in experiment group was much higher than that in control group（P〈0.05-P〈0.01）.Conclusion It suggests that the MSCs infusion can migrate to transplant liver directly and improve the transplant liver function after small-for-size liver transplantation.It is feasible to rescue liver failure after small-for-size liver transplantation with MSCs administration.