【作者】 杨耀国；

【导师】 赵浩亮；

【作者基本信息】 山西医科大学 ， 外科学， 2004， 硕士

【摘要】 目的：探讨同种异体肝细胞经脾脏移植对大鼠急性肝功能衰竭的治疗作用；探讨肝组织中转化生长因子-β1mRNA（TGF-β1mRNA）在肝细胞移植治疗肝功能衰竭时的变化及意义。方法：建立大鼠急性药物性肝衰竭模型：10％D－氨基半乳糖（D-gal）溶液按1.6g/kg一次性腹腔注射；肝细胞的分离和纯化：采用肝脏原位灌注法，制备成肝细胞悬液，肝细胞浓度为4×107个/ml，于2小时内经脾脏移植；动物分组：健康成年Wistar大鼠56只制备成急性肝功能衰竭模型并随机分为两组。Ⅰ组为实验组行脾内肝细胞移植，肝细胞悬液的量为0.5 ml/只，Ⅱ组为对照组在脾内注入0.5ml生理盐水。另外取6只用作分离新鲜肝细胞。术后6小时、12小时、24小时、48小时每组分别取5只检测肝脏功能、肝脏病理改变及应用原位杂交技术检测肝脏组织中TGF-β1 mRNA的变化。结果：肝功能变化：血清丙氨酸氨基转移酶（ALT）6小时和12小时Ⅰ组高于Ⅱ组，差别有统计学意义，P<0.01；ALT在24和48小时Ⅰ组均低于Ⅱ组，差别有统计学意义，P<0.01。总胆红素（TBil）在6小时Ⅰ组高于Ⅱ组，差别有统计学意义, P<0.01；12小时Ⅰ组高于Ⅱ组，差别无统计学意义,在24和48小时Ⅰ组均低于Ⅱ组，差别有统计学意义，P<0.01。肝组织的破坏程度在48小时Ⅰ组比Ⅱ组有较明显的减轻。在各时间点，两组间的白蛋白（ALB）差别无统计学意义。肝脏组织中TGF-β1 mRNA的变化：12小时内两组间无明显区别，24和48小时Ⅰ组均低于Ⅱ组，差别有统计学意义，P<0.01。结论：同种异体肝细胞移植可以改善D-gal诱导的急性肝功能衰竭大鼠的肝脏功能；在24和48小时肝脏组织中的TGF-β1 mRNA Ⅰ组均低于Ⅱ组，可能有利于肝细胞的增殖。更多还原

【Abstract】 Objective：To investigate the effect of hepatocyte transplantation on acute liver failure (ALF) of rats. To investigate the change and significance of transforming growth factor-β1 mRNA ( TGF-β1 mRNA ) in liver after hepatocyte transpl- atation.Methods: 1.Preparation of ALF model. 10% D-gal solution was prepared and injected to rates via abdominal cavity with dosage of 1.6 g/kg respectively. 2.Cell seperation and purification. Collagenase perfusion was performed on six Wistar rats to separate liver cells and suspension was counted . 3.Hepatocyte transplantation experiments. Fifty-six of Wistar rats with ALF induced by D-gal were randomly divided into two groups: Group I and II. Group I was performed intrasplenic hepatocyte transplantation (approximately 2 ×107 cells ). Group II was performed intrasplenic injection of normal saline of same volume. 4.Post-transplantation liver function test. The blood samples of each group of experimental rates were drawn on the 6h , 12h , 24h and 48h after transplantation. ALT and TBil were assayed with automatic biochemical analyzer. Liver sample of rats was performed HE staining and in situ hybridization (ISH). Results： The comparison between group I and II shows that signficant differences were found in ALT and TBil of 24h and 48h( p< 0.01 ). Pathological changes of group I were much mild than its control group at 48h. The comparison between group I and II shows that no signficant differences were found in TGF-β1 mRNA of 6h and 12h(p<0.01). The comparison between group I and II shows that signficant differences were found in TGF-β1 mRNA of 24h and 48h(p<0.01). Conclusion: Intrasplenic transplantation could increase the index of hepatic function of rats with ALF induced by D-gal and hepatic pathology could be <WP=6>improved within 48 hours after transplantation. The decrease of TGF-β1 mRNA in gruop I maybe facilitate to hepatocyte growth.更多还原