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

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250～300 g,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,U=24)或4℃含TAT-HO-1融合蛋白50 μg/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18 h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、18 h时P组保存液AST、ALT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.

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

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 rots. 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 50 μg/ml. Liver specimens and preservation solution samples were collected at 0, 6, 12, and 18 h of cold storage. The activities of aspartato amino-transferase (AST), alanine aminu-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-dependendy 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.