摘要
目的探讨微囊化猪肝细胞移植(HTx)治疗大鼠急性肝衰竭(ALF)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)1．2 g／kg体重腹腔内注射制作SD大鼠ALF模型。ALF大鼠随机分为3组。注药24 h后分别将PRMI 1640培养液2 ml腹腔注射为对照(I组)、2 ml游离猪肝细胞(2×107个／ml)腹腔内移植(Ⅱ组),以2 ml微囊化猪肝细胞腹腔内移植(2×107个／ml,Ⅲ组)。观察移植后大鼠14 d存活率、谷丙转氨酶(ALT)、总胆红素(TB)、血氨(NH3)的改变和肝脏的病理变化。结果肝细胞移植后大鼠14 d存活率:I组为20．0％(5／25),Ⅱ组为66．7％(16／24),Ⅲ组为76．0％(19／ 25),3组间差异有统计学意义(P<0．05)。移植后第1天I组ALT、TB和移植前相比差异无统计学意义,Ⅱ、Ⅲ组ALT、TB下降。第4天3组ALT、TB均继续下降,Ⅱ、Ⅲ组低于I组(P<0．05), Ⅱ、Ⅲ两组间差异无统计学意义(P>0．05)。移植后第7天,ALT进一步下降,3组间差异有统计学意义(P<0．05);TBⅢ组显著低于Ⅱ组和I组(P<0．05),Ⅱ组低于I组但差异无统计学意义 (P>0．05)。移植后第14天,肝功能均明显恢复,ALT、TBⅡ、Ⅲ组均低于I组(P<0．05),Ⅱ、Ⅲ两组间差异无统计学意义(P>0．05)。各组治疗前后NH3有所改善,但各时间段及各组之间差异无统计学意义(P>0．05)。微囊组和游离肝细胞组移植后肝脏病理损害修复较快,而阴性对照组肝脏再生修复较差。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,改善急性肝衰竭大鼠的肝功能,有利于受体受损肝脏的再生修复,但对降低血氮的效果不明显。
Objective To investigate the therapeutic effect of microencapsulated pig hepatocyte transplantation in the treatment of acute liver failure （ALF） in rats. Methods In situ recirculating collagenase perfusion method was used to isolate hepatocytes of Chinese experiment minipigs. ALF in SD rats was induced by D-galactosamine （D-gal） introperitoneal injection （ 1.2 g/kg）. After 24 h of D-gal injection, the ALF rats were divided into three groups. Two ml PRMI 1640 medium was injected into peritoneal cavity of ALF rats （group Ⅰ ）. Two ml free pig hepatocytes containing 2×10^7 pig hepatocytes were transplantated into peritoneal cavity of ALF rats （group Ⅱ ）. Two ml encapsulated pig hepatocytes by alginate-barium （2×10^7/ml） were transplantated into peritoneal cavity of ALF rats （group Ⅲ ）. Fourteenday survival rate, ALT, TB, NH3, and the pathological change of liver were observed. Results There were significant differences in survival rate at 14th day among group Ⅰ （20.0%）, group Ⅱ（66.7%） and group Ⅲ （76.0%） （P〈0.05）.There were no significant differences in ALT and TB in group Ⅰ on the first day compared with pre-transplantation. ALT and TB were declined in group Ⅱ and group Ⅲ after transplantation. ALT and TB were declined in three groups on the 4th day. The indices in group Ⅱ and group Ⅲ were lower than those in group Ⅰ （P 〈 0.05）, but there was no significant difference between group Ⅱ and group Ⅲ （P 〉0.05）. ALT was declined further on the 7th day. There were significant differences in ALT among three groups （ P 〈 0.05）. TB in group Ⅲ was lower than that in group Ⅱ and group Ⅰ （ P 〈 0.05）, but there was no significant difference in TB between group Ⅱ and group Ⅰ （P 〉0.05）. Up to the 14th day, hepatic function was improved remarkably, and ALT and TB in group Ⅱ and group Ⅲ were lower than those in group Ⅰ （P 〈 0.05）, But there were no significant differences in ALT and TB between group Ⅱ and group Ⅲ （P〉0.05）. In three groups, NH3 of rats after transplantation was lower than that pre-transplantation, but there were no differences in NH3 before and after transplantion （P 〉 0.05）. The liver pathological damage was quickly improved in group Ⅱ and group Ⅲ as compared with group Ⅰ . Conclusion After microencapsulated pig hepatocytes transplantation, the survival rate of ALF rats induced by D-gal was increased, and their hepatic function and regeneration were also improved. But microencapsulated pig hepatocytes transplantation could not reduce blood NH3 significantly.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2005年第12期1469-1471,共3页
Chinese Journal of Experimental Surgery
基金
江苏省卫生厅2002年重大项目基金资助项目(H200215)
关键词
肝衰竭
微囊
肝细胞
移植
猪
细胞移植
大鼠
Liver failure
Microencapsulate
Hepatocytes
Transplantation
作者简介
通讯作者：陈钟
