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目的为了明确血红素加氧酶-1的高表达对大鼠减体积肝移植生存的影响,进而探索出一条有效地防止小体积移植肝缺血再灌注损伤的方法。方法利用分子生物学方法构建携带有HO-1基因的重组腺病毒Ad5-HO-1,对供体进行预处理,并观察Ad5-HO-1对大鼠小体积移植肝缺血再灌注损伤的保护作用。结果Ad5-HO-1组与对照组相比较ALT水平有显著降低(P＜0．05)；术后2 h内门静脉血流流量有明显增加(P＜0．05)；HO-1酶活性测定显示Ad5-HO-1组酶活性为(5．34±0．41)×10~(-3)mmol·mg~(-1)·mm~(-1),较对照组明显为高(P＜0．05)；RT-PCR和免疫组化检测HO-1、TNF-α、Bcl-2、Bax表达,证实Ad5-HO-1组中HO-1、Bcl-2的表达是增高的,而Bax、TNF-α表达是降低的(P＜0．05)；Ad5-HO-1组在1、7、21d的生存率分别为100%、85．7%、71．4%,而对照生理盐水组在1、7、21 d的生存率分别为66．7%、16．7%、16．7%(P＜0．05)。结论Ad5-HO-1的应用能够显著地增加术后2 h内门静脉血流,有利于受体术后肝功能的恢复,从而改善大鼠移植术后的生存结果。实验结果提示HO-1的高表达在防止和减轻小肝移植物缺血再灌注损伤的过程中扮演着重要的角色,进而为临床上防止小肝移植物缺血再灌注损伤提供了一条新的思路和方法。
Objective To find out the effect of high expression of heme oxygenase-1 on the survival of rat with reduced-size liver transplantation, and to find out an effective way to prevent liver ischemia-reperfusion injury in small volume transplantation. Methods The recombinant adenovirus Ad5-HO-1 carrying the HO-1 gene was constructed by molecular biology method. The donor was pretreated with Ad5-HO-1 and the effect of Ad5-HO-1 on the hepatic ischemia-reperfusion injury Protective effects. Results The level of ALT in Ad5-HO-1 group was significantly lower than that in control group (P <0.05), and the portal vein blood flow increased significantly within 2 hours after operation (P <0.05). The activity of HO-1 The enzyme activity of Ad5-HO-1 group was (5.34 ± 0.41) × 10 -3 mmol · mg -1 (-1) mm -1, which was significantly higher than that of the control group P <0.05). The expression of HO-1, TNF-α, Bcl-2 and Bax were detected by RT-PCR and immunohistochemistry. The expression of HO-1 and Bcl-2 in Ad5-HO- , While the expression of Bax and TNF-α was decreased (P <0.05). The survival rates of Ad5-HO-1 group were 100%, 85.7% and 71.4% The survival rates of control saline group at 1, 7, 21 days were 66.7%, 16.7% and 16.7%, respectively (P <0.05). Conclusion The application of Ad5-HO-1 can significantly increase the portal vein blood flow within 2 h after operation, which is beneficial to the recovery of postoperative liver function and thus improve the survival outcome after transplantation. The experimental results suggest that the high expression of HO-1 plays an important role in preventing and reducing the ischemia-reperfusion injury of small liver graft, and thus provides a new Ideas and methods.