目的建立大鼠小体积肝移植模型,输注表达人肝细胞生长因子(human hepatocyte growth factor,hHGF)的骨髓间充质干细胞(mesenchymal stemcells,MSCs),研究其在移植早期对小移植肝促再生作用。方法将已建立的表达hHGF和绿色荧光蛋白(green fluorescence protein,GFP)的MSCs,分别命名为HGF/MSCs,GFP/MSCs。建立大鼠30%肝移植模型。受体分为4组,实验组输注5×10^6HGF/MSCs;对照组则分别输注相同体积的生理盐水(PS),5×10^6GFP/MSCs或1.0×10^9pfu含hHGF的重组腺病毒液(Ad-HGF)。分别于术后1,3,5,7d各组随机抽取5只大鼠处死。取血检测血清ALT和hHGF。记录移植物湿重。取肝组织检测hHGF、c-met表达,以及肝细胞凋亡和增殖活性。另每组15只,分组同上,用于观察生存期。结果PS组大鼠7d生存率33.3%;组织学及血清学检查示术后肝脏损伤重,汇管区单核细胞浸润多;而实验组大鼠7d生存率为73.3%,肝脏损伤轻,炎性细胞浸润少;实验组移植肝再生较PS组明显增加。结论大鼠部分肝移植后,输注HGF/MSCs能够保护小体积移植肝,促进小移植肝再生,提高7d生存率。
Objective To determine whether HGF-expressing MSCs will improve small for-size liver grafts regeneration after liver transplantation. Methods The HGF cDNA was amplified from the expression plasmid pCMV-HGF by PCR and subcloned into the same site of the plasmid pDC316- IRES-EGFP vector. Virus Ad-HGF was produced by homologous recombination. BM-MSCs were cul tured and transduced with Ad-HGF. HGF/MSCs were generated. We implanted HGF/MSCs into liver grafts via the portal vein using a 30% small-forsize rat liver transplantation model. HGF, c-met expression, hepatic injury and liver regeneration were assessed after liver transplantation. Results The HGF-expressing MSCs improved liver function and liver weight recovery during the early post transplantation period, resulting in a reduced mortality in rats after small-for-size liver transplantation. Conclusions The MSCs genetically modified to over-expressing HGF and implanted in the liver graft may offer a novel approach to promoting liver regeneration after small-for-size transplantation.