【摘要】 探讨同种异体肝细胞经腹腔、脾脏移植对D 氨基半乳糖 (D gal)诱导的急性肝衰竭大鼠的治疗作用及移植细胞的生物学特性 ;研究基因修饰肝细胞经脾脏移植后的定位及表达。方法　采用D gal诱导大鼠肝衰竭。在D gal诱导后 4 8h ,Ⅰ组 :经腹腔移植 4× 10 7同种异体肝细胞 ;Ⅱ组 :经腹腔注射生理盐水 2ml;Ⅲ组 :经脾脏移植 2× 10 7同种异体肝细胞 ,同时肌注CsA 10mg/kg ;Ⅳ组 :经脾脏注射生理盐水 1ml,同时肌注CsA 10mg/kg ,观察大鼠的存活率、肝脏功能、肝脏病理变化和移植细胞的组织学及G6P活性变化 ;Ⅴ组 :经脾脏移植双顺反子逆转录病毒修饰的大鼠原代肝细胞 ,同时肌注CsA 10mg/kg ,经PCR扩增及X gal染色方法追踪细胞的定位及基因表达。 结果　Ⅰ组大鼠存活率显著高于Ⅱ组 ( 71.4 %对 2 6.7% ,P <0 .0 5) ,肝脏功能及肝脏病理有部分改善。腹腔内移植肝细胞可短暂存活并具有G6P活性。Ⅲ组大鼠存活率与Ⅳ组无差别 ( 30 %对 30 % ,P >0 .0 5) ,肝功能、肝脏病理无明显改善。脾内移植肝细胞可短暂存活并具有G6P活性。Ⅴ组经脾移植基因修饰肝细胞后 ,可在脾内检测到 β gal基因表达并扩增出NeoR基因。 结论　经腹腔移植同种异体肝细胞可改善D gal诱导的肝衰竭大鼠的存活率 ,部分改善肝功能及肝脏病理 ,移植细胞更多还原

【Abstract】 Objective Study on the effect of allogeneic hepatocyte transplantation (HcT) intraperitoneally or intrasplenically in rats with acute hepatic failure (AHF) which were induced by D galactosamine, and to study the cell location and gene expression after retrovirus infected hepatocyte transplantation. Methods AHF animals were induced by D gal. HcT was received 48h after intoxication. Group Ⅰ received 4×10 7 hepatocytes intraperitoneally. Group Ⅱ received 2mL N.S intraperitoneally. Group Ⅲ received intrasplenic injection of 2×10 7 hepatocytes with Cyclosporin A(CsA) 10mg/kg simultaneously. Group Ⅳ received intrasplenic injection of 1 mL N.S with CsA 10mg/kg. The survival rates, liver function, liver histology and biologic characteristics of transplanted cells were assessed. Group Ⅴ received intrasplenic transplantation of hepatocytes infected with bicistronic retrovirus containing NeoR gene and β galactosidase (β gal) gene. The transplanted hepatocytes were traced through PCR amplication and X gal staining.Results Survival rate of group Ⅰ was higher than that of group Ⅱ (71.4% vs 26.7%, P <0.05), the liver function and liver histology were partially improved in group Ⅰ as compared with group Ⅱ. The intraperitoneally transplanted hepatocytes could survive and show G6P activity temporarily. Survival rate of group Ⅲ was the same as that of group Ⅳ (30% vs 30%, P >0.05). Liver function and liver histology of group Ⅲ were not improved significantly as compared with group Ⅳ, but the hepatocytes could temporarily survive and show G6P activity in the splenic parenchyma. The AHF rats in group Ⅴ expressed β gal gene in the spleen and NeoR gene were amplified in the spleen.Conclusion Survival rate of AHF rats intoxicated with D gal could be improved, the liver function and liver histology could also be partially improved, via intraperitoneal HcT. The intraperitoneally transplanted hepatocytes could temporarily survive and show specific hepatocyte function, whereas the survival rate, liver function and liver histology of AHF rat through intrasplenic HcT could not be improved, the transplanted hepatocytes could temporarily survive and show specific hepatocyte function in the spleen. The gene modified hepatocytes transplanted into splenic parenchyma were located in the spleen and expressed β gal gene.更多还原