微囊化猪肝细胞移植对急性肝衰竭大鼠肝功能支持的实验研究

The therapeutic effect of microencapsulated pig hepatocytes transplantation on rats with acute liver failure

目的:探讨微囊化猪肝细胞移植治疗大鼠急性肝衰竭的效果。方法:采用原位胶原酶循环肝灌注法分离中国实验用小型猪肝细胞,海藻酸钠-氯化钡法微囊化。应用D-氨基半乳糖(D-gal)1.2g/kg体重腹腔内注射制作SD大鼠急性肝衰竭模型。急性肝衰竭大鼠随机分为3组。注药24h后分别将PRMI1640培养液2ml腹腔注射为对照组(1组)、2ml游离猪肝细胞(2×107个/ml)腹腔内移植组(2组)、2ml微囊化猪肝细胞(2×107个/ml)腹腔内移植组(3组)。观察移植后大鼠14天存活率、谷丙转氨酶(ALT)、总胆红素(TB)的改变。结果:肝细胞移植后大鼠14天存活率:1组为20.0%(5/25),2组为66.7%(16/24),3组为76.0%(19/25),3组间差异有显著性(P<0.05)。移植后第1天1组ALT、TB和移植前相比差异无显著性,2、3组ALT、TB下降。第4天3组ALT、TB均继续下降,2、3组低于1组(P<0.05),2、3两组间差异无显著性(P>0.05)。移植后第7天,ALT进一步下降,3组间差异有显著性(P<0.05);TB3组显著低于2组和1组(P<0.05),2组低于1组但差异无显著性(P>0.05)。移植后第14天,肝功能均明显恢复,ALT、TB2、3组均低于1组(P<0.05),2、3两组间差异无显著性(P>0.05)。结论:微囊化猪肝细胞腹腔内移植可以提高药物诱导急性肝衰竭大鼠的存活率,改善ALF大鼠的肝功能。

Objective:To investigate the therapeutic effect of microencapsulated pig hepatocyte transplantation on acute liver failure rats.Methods:In situ recirculating collagenase perfusion method was used to isolate hepatocytes of Chinese experiment minipigs.Acute liver failure in SD rats was induced by D-galactosamine (D-gal) introperitoneal injection (1.2g/kg).After 24h of D-gal injection, the ALF rats were divided into three groups. 2ml PRMI 1640 medium was injected into peritoneal cavity of ALF rats (group 1). 2ml free pig hepatocytes containg 4×107 pig hepatocytes were transplantated into peritoneal cavity of ALF rats (group 2).2ml encapsulated pig hepatocytes by alginate-barium(2×107/ml) were transplantated into peritoneal cavity of ALF rats (group 3).14d survival rate,ALT,TB,were observed.Results:There were significant differences in survival rate at 14th day among group 1(20.0%), group 2(66.7%) and group 3 (76.0%) (P<0.05).There were no significant differences in ALT and TB in group 1 on first day compared with pre-transplantation.ALT and TB declined in group 2 and group 3 after transplantation.ALT and TB declined in three groups on 4th day.The indices in group 2 and group 3 were lower than those in group 1(P<0.05).But there was no significant difference between group 2 and group 3(P>0.05).ALT declined further on 7th day. There were significant differences in ALT of three groups (P<0.05).TB in group 3 was lower than that in group 2 and group 1(P0.05).But there was no significant difference in TB between group 2 and group 1(P>0.05).Up to 14th day, hepatic function improved remarkably,ALT and TB in group 2 and group 3 was lower than those in group 1(P<0.05).But there were no significant differences in ALT and TB between group 2 and group 3(P>0.05).Conclusions:After microencapsulated pig hepatocytes transplantation,the survival rate of ALF rats induced by D-gal increased.