急性肝衰竭小鼠necroptosis的发生及Nec-1的干预作用研究

Study on the occurrence of programmed necrosis and the intervention of inhibitors in acute liver failure

目的分析急性肝衰竭小鼠中程序性坏死(necroptosis)发生情况及特异性抑制剂-1(Nec-1)的干预作用。方法 2017年6月—2018年3月在解放军总医院第五医学中心进行动物实验。60只SPF级雄性C57/BL6小鼠随机数字表法分为对照组、模型组(D-半乳糖胺和脂多糖造模)、干预组(造模前给予Nec-1)每组20只,检测小鼠造模后肝功能指标(ALT)、炎性因子(TNF-α、IL-6、IL-10)、受体相互作用蛋白(RIP),并观察肝脏病理变化。结果造模后1 h,急性肝衰竭模型组ALT、TNF-α、IL-6、IL-10、RIP1、RIP3水平均明显高于对照组(t/P=6.821/<0.001,11.386/<0.001,10.609/<0.001,8.541/<0.001,5.867/<0.001;4.388/0.002),干预组的ALT、TNF-α、IL-6、IL-10水平均低于模型组(t/P=2.753/0.023,3.558/0.012,5.796/<0.001,3.332/0.011);3 h时干预组的RIP1水平低于模型组(t=2.416,P=0.04),其余时间点以及RIP3水平比较2组差异无统计学意义(P>0.05)。结论程序性坏死在急性肝衰竭的发生中发挥了作用,Nec-1可在一定程度上抑制其作用。

Objective To analyze the occurrence of programmed necrosis in acute liver failure and the intervention effect of programmed necrosis specific inhibitor 1(Nec 1).Methods Animal experiments were conducted in the Fifth Medical Center of PLA General Hospital from June 2017 to March 2018.Sixty C57/BL6 mice were randomly divided into normal control group,model group(D galactosamine and lipopolysaccharide modeling),intervention group(Nec 1 before modeling),20 mice in each group.Liver function index(ALT),inflammatory factors(TNF-α,IL-6,IL-10),receptor interaction protein(RIP)were detected after modeling,and liver pathological changes were observed.Results At 1 h after model establishment,the levels of ALT,TNF-α,IL-6,IL-10,RIP1 and RIP3 in the acute liver failure model group were significantly higher than those in the normal control group(t=6.821,P<0.001;t=11.386,P<0.001;t=10.609,P<0.001;t=8.541,P<0.001;t=5.867,P<0.001;t=4.388,P=0.002),the ALT,TNF-α,IL-6 and IL-10 levels in the intervention group were lower than Model group(t=2.753,P=0.023;t=3.558,P=0.012;t=5.796,P<0.001;t=3.332,P=0.011);RIP1 level of Nec 1 intervention group was lower than model group at 3 h(t=2.416,P=0.04),there was no significant difference between the other time points and the RIP3 level(P>0.05).Conclusion Programmed necrosis plays a role in the development of acute liver failure,and Nec-1 can inhibit its effects to some extent.