摘要
目的探讨HIV-1反式激活蛋白（transactiviting protein，TAT）与血红素氧合酶-1（heme oxygenase，HO-1）融合蛋白（TAT-HO-1）对大鼠供肝冷保存期肝细胞凋亡的影响。方法选择雄性SD大鼠48只，体重250g-300g。根据所用保存液的不同随机分为两组（n=24）：C组，肝脏应用4℃HTK液进行灌注和冷保存；P组，肝脏应用4℃含TAT-HO-150mg／L的HTK液进行灌注和冷保存。参照Kamada等的方法切取肝脏，分别于冷保存0、6、12、18h后，收集保存液，切取肝脏组织。免疫组化染色法分析TAT-HO—1转导入肝脏的情况，全自动生化分析仪测定保存液中谷丙转氨酶（alanine aminotransferase，ALT）水平，放射免疫分析法检测肝组织中透明质酸（hyaluronic acid，HA）水平，TUNEL法检测肝实质细胞和肝窦内皮细胞（sinusoidal endothelial cell，SECs）凋亡指数，免疫组化染色法检测肝组织Caspase-3表达情况。结果免疫组化染色结果显示P组肝组织中HO-1含量在冷保存6、12、18h时均高于C组（P〈0．05）。两组保存液中ALT水平，肝组织中HA含量，肝实质细胞和SECs凋亡率以及肝组织Caspase-3表达水平均随保存时间延长而升高（P〈0．05）。冷保存6、12、18h后保存液中ALT水平、肝组织HA含量、肝实质细胞、SECs凋亡指数以及Caspase-3阳性表达水平P组均低于C组（P〈0．05）。结论TAT-HO-1可以在冷保存期间转导进入大鼠肝脏并减轻肝实质细胞和SECs凋亡，对肝脏功能具有一定保护作用。蛋白转导技术将成为减轻肝移植冷保存期损伤的新措施。
Objective To study the effects of TAT-HO-1 fusion protein, HIV-1 transactiviting protein （TAT） and heine oxygenase-1 （HO-1）, on hepatic cell apoptosis of rat donors in cold storage stage. Methods Forty-eight male SD rats were randomly divided into two groups. Rat livers were flushed and preserved with 4℃ HTK solution containing（group P） or uncontaining（group C） 50 mg/L of TAT-HO-1. The preserved solution and hepatic tissue were collected at 0, 6, 12, 18 h of cold storage stage. TAT-HO-1 transducing into liver, alanine aminotransferase（ALT） level in preserved solution, hyaluronic acid（HA） level and the expression of caspase-3 in hepatic tissue, and the apoptotic index （AI） of hepatocytes and sinusoidal endothelial cells （SECs） were measured or detected. Results ALT level in preserved solution, HA level and the expression of caspase-3 in hepatic tissue, and the AI of hepatocytes and SECs increased time-dependently in cold storage stage in both groups （P〈0.05）, with lower increasing extent in group P than that in group C （P〈0.05） at 6h, 12h and 18h of cold storage stage. A stronger accumulation of HO-1 staining was also detected at the same time-points in group P than that in group C（P〈0.05）. Conclusion TAT-HO-1 may transduce efficiently into rat livers, exerting protective effects on both hepatocytes and SECs during cold storage stage. Protein transduction technology may be a novel therapeutic means to reduce donor liver injury in preservation period for transplantion.
出处
《国际麻醉学与复苏杂志》
CAS
2009年第4期328-331,共4页
International Journal of Anesthesiology and Resuscitation
基金
国家自然科学基金资助项目（30672024）
