表达HGF的骨髓间充质干细胞促进小移植肝早期肝再生

Improvement of small-for-size liver graft regeneration by bone marrow-derived mesenchymal stem cellsover-expressing hepatocyte growth factor

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.