RIP3在大鼠肝脏缺血后处理中的表达与作用

摘要

Objective:To investigate the expression and function of RIP3 in the liver of rats following ischemic postconditioning.Methods:The model of 70％ hepatic ischemia and reperfusion was established,then a total of forty healthy adult male Sprague-Dawley(SD) rats were divided randomly into four groups,ten rats in each group:a sham-operation group (Sham group);an ischemia reperfusion injury group(IR group);an ischemic postconditioning group(IPO group);an ischemic postconditioning and necrostatin-1 group (Nec-1 group).The blood samples and liver tissues were collected.The serum levels of ALT and AST were detected,and the liver histological examination was performed.Western-bolt was used to detect the TNF-α and RIP3 levels.Results:Compared with the IR group,ALT and AST in serum were significantly declined in the IPO group (P＜0.05);The liver damage after ischemia and reperfusion was improved obviously in the IPO group compared to which in the IR group;The Suzuki's scores was increased in the IR group compared to which in the IPO group (P＜0.05);There was a low grade of TNF-α and RIP3 in the Sham group,whereas the level of TNF-α and RIP3 significantly increased in the IR and IPO and Nec-1 group(P＜0.05);Compared with the IR group,the level of RIP3 was further decreased in the IPO group (P＜0.05);Compared with the IPO group,the level of RIP3 was further decreased in the Nec-1 group (P＜0.05).Conclusion:RIP3-mediated necroptosis was involved during hepatic ischemia postconditioning,and the protective effect of ischemia postconditioning may act as reducing necroptosis by cutting down the levels of RIP3.%目的:探讨受体相互作用蛋白3(RIP3)在大鼠肝脏缺血后处理中的表达与作用.方法:采用大鼠70％肝脏热缺血再灌注损伤模型,40只健康成年雄性SD大鼠随机分为4组:假手术组(Sham组)、缺血再灌注损伤组(IR组)、缺血后处理组(IPO组)和缺血后处理+程序性坏死特异性抑制剂-1组(Nec-1组).取下腔静脉血检测ALT和AST的水平;HE染色观察组织病理学改变并进行Suzuki's评分;蛋白免疫印迹法(Western blot)检测肝组织中肿瘤坏死因子-α(TNF-α)和RIP3的蛋白表达水平.结果:与IR组相比,IPO组肝组织病理学表现明显改善,Suzuki's评分明显降低(P＜0.05),血清ALT、AST水平明显下降(P＜0.05);与Sham组相比,IR组、IPO组及Nec-1组TNF-α和RIP3表达水平均明显升高(P＜0.05);与IR组相比,IPO组RIP3表达水平显著降低(P＜0 05);与IPO组相比,Nec-1组RIP3表达水平明显降低(P＜0.05).结论:肝脏缺血后处理中有RIP3介导的程序性坏死的参与,缺血后处理的保护机制可能与降低RIP3的表达从而减少程序性坏死的发生有关.