梗阻性黄疸SD大鼠肝热缺血再灌注损伤模型的建立

Rat model of hepatic warm ischemia reperfusion injury with obstructive jaundice

目的构建阻塞性黄疸合并肝热缺血再灌注损伤的SD大鼠模型。方法选择SD大鼠40只随机分为4组:假手术组(Sham组)、梗阻7天组(7 d组)、梗阻14天组(14 d组)及梗阻21天组(21 d组),每组10只。通过双重结扎并切断胆总管建立SD大鼠阻塞性黄疸模型,探索最佳梗阻时间。梗阻7 d后另择SD大鼠62只,再随机分为4组:未缺血组(0 min组,10只)、缺血15 min组(15 min组,12只)、缺血30 min组(30 min组,16只)、缺血45 min组(45 min组,24只),分别行肝门部血管阻断0、15、30、45 min后再灌注,建立后续肝热缺血再灌注损伤模型,观察生化指标、生存率及病理学改变。结果梗阻7 d时适合下一步建模。建立阻塞性黄疸模型后,随着肝门部血管阻断时间的延长,肝功能进行性下降,大鼠死亡率逐渐升高,各组间的差异具有统计学意义(P<0.05);随阻断时间的延长,肝脏损害的病理学表现更为严重。其中肝门部血管阻断30 min时24 h死亡率50%,再灌注2 h后病理学上出现严重的肝细胞变性、坏死改变。结论对于梗阻性黄疸的SD大鼠,在梗阻7 d后行肝门部血管阻断30 min再恢复血流,可有效建立梗阻性黄疸大鼠的肝缺血再灌注损伤模型。

Objective To establish the model of hepatic warm ischemia reperfusion injury with obstructive jaundice on sprague-dawley rats. Methods One hundred and two mature SPF sprague-dawley rats were used in the establishment. Among them 40 rats were randomly divided into 4 groups： sham group, 7 days obstructive jaundice group, 14 days obstructive jaundice group and 21 days obstructive jaundice group, with 10 rats in each group. Double ligation and cutting the common bile ducts in the 7, 14, and 21 d groups were used to investigate the optimal time of obstruction. The other 62 rats were randomly divided into 4 groups after 7 days of obstruction：non-ischemia group （0 min group, n=10）, 15 min ischemia group （n=12）, 30 min ischemia group （n=16） and 45 min ischemia group （n=24）. Pringle method of blocking hepatic portal vessels was used for 0, 15, 30 and 45 min correspondingly. Liver function, survival rate and pathological changes were observed respectively. Results The model of hepatic warm ischemia reperfusion injury was better established after obstruction for 7 days on sprague-dawley rat. After 7 days of obstructive jaundice, along with the prolongation of Pringle method of blocking hepatic portal vessels, the liver function decreased progressively and the mortality rate increased gradually. There were statistical differences between the 4 groups （P〈0.05）. The hepatic pathological changes such as the degeneration and necrosis of hepatic cells were more severe gradually with the ischemia time going. The＆nbsp;mortality rate was 50%after 30 min hepatic portal vessels blocking on 7 days obstructive jaundice rats, and there was serious degeneration and necrosis of hepatic cells after 2 hours of reperfusion. Conclusion Obstruction for 7 days on sprague-dawley rats and then blocking hepatic portal vessels for thirty minutes can effectively establish an appropriate model of obstructive jaundice and hepatic warm ischemia reperfusion injury rats.