TAT-血红素氧合酶-1融合蛋白对原位肝移植术大鼠肝损伤的影响

Effects of TAT-heme oxygenase-1 fusion protein on liver injury in rats undergoing orthotopic liver transplantation

目的 评价TAT-血红素氧合酶-1（TAT-HO-1）融合蛋白对原位肝移植术大鼠肝损伤的影响.方法 选择健康成年雄性近交系Lewis大鼠为供体,BN大鼠为受体,8～10周龄,体重180～230 g.采用随机数字表法,将受体大鼠分为2组（n=6）：肝移植组（OLT组）和TAT-HO-1融合蛋白组（TAT-HO-1组）.采用改良Kamada二袖套法建立原位肝移植术模型.TAT-HO-1组植入经含TAT-HO-1融合蛋白50μg/ml的HTK液冷保存6h的肝脏.于移植后7d,经腹主动脉采血样2 ml,测定血清ALT和AST的活性.取肝脏,HE染色,光镜下观察肝组织病理学结果,计算排斥活动指数.采用ELISA法检测肝组织转化生长因子-β1（TGF-β1）及IL-6的含量.分离Kupffer细胞培养,测定培养液TGF-β1及IL-6的水平.结果 与OLT组比较,TAT-HO-1组血清ALT及AST的活性、排斥活动指数、肝组织和Kupffer细胞培养液TGF-β1及IL-6的水平降低（P＜0.05）,肝组织病理学损伤减轻.结论 在供肝冷保存期间,应用TAT-HO-1融合蛋白可减轻原位肝移植术大鼠肝损伤.

Objective To evaluate the effects of TAT-heme oxygenase-1 （TAT-HO-1） fusion protein on liver injury in rats undergoing orthotopic liver transplantation （OLT）.Methods Adult male Lewis （inbred） rats （aged 8-10 weeks,weighing 180-230 g） were used as donors and Brown Norway rats （aged 8-10 weeks,weighing 180-230 g） as recipients.The recipient rats were randomly divided into 2 groups （n=6 each） using a random number table：OLT group and TAT-HO-1 group.The livers were harvested according to the method described by Kamada.In OLT group,the donor livers were flushed and preserved with 4 ℃ HTK solution,while the livers were flushed and preserved for 6 h with 4 ℃ HTK solution containing TAT-HO-1 50 μg/ml in group P.Blood samples were obtained at 7 days after transplantation for measurement of activities of alanine aminotransferase and aspartate aminotransferase in serum.Hepatic specimens were obtained at 7 days after transplantation and stained with haematoxylin and eosin for examination under light microscope.Rejection activity index was calculated according to Banff criteria.The contents of transforming growth factor-beta 1 and interleukin-6 in liver tissues were determined using ELISA.Kupffer cells were isolated and cultured for 48 h to determine the levels of transforming growth factor-beta 1 and interleukin-6 in culture medium.Results Activities of alanine aminotransferase and aspartate aminotransferase in serum,rejection activity index and levels of transforming growth factor-beta 1 and interleukin-6 in liver tissues and culture medium of Kupffer cells were significantly decreased,and the pathological changes of livers were mitigated in group TAT-HO-1 as compared to group OLT.Conclusion TAT-HO-1 fusion protein applied during cold storage of donor livers can attenuate liver injury in rats undergoing OLT.