TAT-血红素氧合酶-1融合蛋白对大鼠供肝冷保存期炎性反应及肝功能的影响

Effects of TAT-heine oxygenase-1 fusion protein on inflammatory response and liver function during liver cold preservation in rats

目的探讨HIV-1反式激活蛋白-血红素氧合酶-1（TAT-HO-1）融合蛋白对大鼠供肝冷保存期炎性反应及肝功能的影响。方法成年健康雄性SD大鼠48只，体重250-300g，开腹游离腹主动脉并结扎，于左右髂总动脉分叉处向心端穿刺腹主动脉，切断肝上下腔静脉，随机灌注4℃ HTK液（C组，n=24）或4℃含TAT-HO—1融合蛋白50μg/ml的HTK液（P组，n=24），至流出灌洗液清亮后停止灌注，取肝脏，置于相应保存液中冷保存。分别于冷保存即刻、6、12和18h时取保存液并切取肝组织。采用全自动生化分析仪测定保存液谷草转氨酶（AST）、谷丙转氨酶（ALT）及乳酸脱氢酶（LDH）活性，免疫组化法检测肿瘤坏死因子α（TNF-α）的表达，光镜下观察肝组织形态学。结果随冷保存时间延长，两组保存液AST、ALT及LDH活性升高，TNF-α表达上调（P〈0．05）；与C组比较，冷保存6、12、18h时P组保存液AST、AST及LDH活性降低，TNF-α表达下调（P〈0．05）。结论TAT-HO-1融合蛋白可抑制大鼠供肝冷保存期的炎性反应，对肝功能具有一定保护作用。

Objective To investigate the effect of TAT-heine oxygenase-1 （TAT-HO-1） fusion protein on the inflammatory response and liver function during cold preservation of liver in rats. Methods Forty-eight male SD rats weighing 250-300 g were randomly divided into 2 groups according to the type of preservation solution （n =24 each）： control group （group C） and TAT-HO-1 fusion protein group （group P）. The livers were harvested according to the method described by Kamada. In control group the livers were flushed and preserved with 4℃ HTK solution while in group P the livers were flushed and preserved with 4℃ HTK solution containing TAT-HO-1 fusion protein 50μg/ml. Liver specimens and preservation solution samples were collected at 0, 6, 12, and 18 h of cold storage. The activities of aspartate amino-transferase （AST）, alanine amino-transferase （ALT） and lactate dehydrogenase （LDH） in the preservation solution were analyzed. TNF-α expression in the liver was determined. The liver histomorphology was observed by light microscopy. Results The AST, ALT and LDH activities in the preservation solution and TNF-α expression in the liver were significantly increased time- dependently during cold storage in both groups. The AST, ALT and LDH activities and TNF-α expression were significantly lower at 6, 12 and 18 h of cold storage in group P than in group C. Conclusion TAT-HO-1 fusion protein can reduce the inflammatory response of the liver during cold storage in rats, and has protective effect on liver function to some extent.