阿托品联合新斯的明减轻肝脏缺血再灌注损伤作用及其机制研究

Effect of Atropine Combined with Neostigmine on Hepatic Ischemia-reperfusion Injury and Its Mechanism

目的:研究阿托品联用新斯的明减轻肝脏缺血再灌注损伤作用及机制,为临床抗缺血再灌注损伤提供可能方案。方法:将C57BL/6J小鼠分为对照组、模型组和阿托品及新斯的明预处理组(0.1 mg·kg^-1+0.2 mg·kg^-1、0.2 mg·kg^-1+0.4mg·kg^-1)。除对照组外,其余各组小鼠建立肝脏缺血1h再灌注6h损伤模型。治疗组于缺血前30 min腹腔注射阿托品及新斯的明,对照组和模型组给予等量生理盐水。于再灌注6h后取缺血肝组织进行病理学检查,采用试剂盒检测缺血肝叶的caspase-3、caspase-8、caspase-9、超氧化物歧化酶(SOD)活性、丙二醛(MDA)、活性氧簇(ROS)及过氧化氢(H2O2)含量变化。结果:病理学检查表明再灌注6 h后小鼠肝脏损伤严重,凋亡肝细胞数量增多,阿托品及新斯的明预处理后能明显减轻缺血再灌注所致的肝损伤;与对照组比较,模型组caspase-3、caspase-8、caspase-9、SOD活性、MDA、ROS及H2O2含量显著升高(P<0.05或P<0.01);与模型组比较,阿托品及新斯的明预处理组(0.2 mg·kg^-1+0.4 mg·kg^-1)caspase-3、caspase-8、caspase-9、MDA、ROS及H2O2含量显著降低,SOD活性显著升高(P<0.05或P<0.01)。结论:阿托品联合新斯的明预处理可减轻肝脏缺血再灌注损伤。

Objective:To study the effect and mechanism of atropine combined with neostigmine in alleviating hepatic ischemia-reperfusion injury to provide a possible scheme for clinical anti-ischemia-reperfusion injury.Methods:C57BL/6J mice were divided into the control group,the model group,atropine and neostigmine pretreatment groups(0.1 mg·kg^-1+0.2 mg·kg^-1,0.2 mg·kg^-1+0.4 mg·kg^-1).Except for the control group,the other two groups were induced by hepatic ischemia for 1 h and reperfusion for 6 h.The treatment group was intraperitoneally injected with atropine and neostigmine 30 minutes before ischemia,while the control group and the model group were given the same amount of saline.After the 6-h reperfusion,ischemic liver tissue was withdrawn for pathological examination,and the changes of caspase-3,caspase-8,caspase-9,superoxide dismutase(SOD),malondialdehyde(MDA),reactive oxygen species(ROS)and hydrogen peroxide(H2O2)in ischemic liver were detected by kits.Results:The pathological examination showed that the hepatic injury was serious and the number of apoptotic hepatocytes increased after the 6-h reperfusion,and atropine and neostigmine pretreatment could significantly alleviate the hepatic injury induced by ischemia-reperfusion.Compared with those in the control group,caspase-3,caspase-8,caspase-9,SOD activity,MDA,ROS and H2O2 contents in the model group increased significantly(P<0.05 or P<0.01).Compared with those in the model group,the contents of caspase-3,caspase-8,caspase-9,MDA,ROS and H2O2 in atropine and neostigmine pretreatment group(0.2 mg·kg^-1+0.4 mg·kg^-1)decreased significantly,while the activities of SOD increased significantly(P<0.05 or P<0.01).Conclusion:Atropine combined with neostigmine pretreatment can alleviate hepatic ischemia-reperfusion injury.