清胰汤对肝缺血再灌注损伤小鼠的肝脏保护作用及铁死亡机制研究  

Study on the Protective Effect of Qingyi Decoction on Hepatic Ischemia-reperfusion Injury in Mice and the Mechanism of Ferroptosis

作　　者：周桢 凌琪华[1] 王倩[1] 方荣[1] Zhou Zhen;Ling Qihua;Wang Qian;Fang Rong(Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院,上海201203

出　　处：《中国中医急症》2024年第1期27-31,共5页Journal of Emergency in Traditional Chinese Medicine

基　　金：上海市科学技术委员会科技创新行动计划(21Y11920500)。

摘　　要：目的 观察清胰汤对肝缺血再灌注损伤小鼠肝脏的保护作用并探讨其机制。方法 30只雄性SPF级C57BL/6小鼠随机分为5组:假手术组、模型组及清胰汤低、中、高剂量组,每组6只。各清胰汤组小鼠于造模前7 d连续灌胃相应剂量药物,假手术组及模型组小鼠灌胃生理盐水。以夹闭小鼠肝左叶及肝中叶动静脉血流的方法构建70%肝脏缺血再灌注损伤模型,缺血60 min,再灌注12 h。以ELISA法检测血清肝酶指标;HE染色观察肝组织缺血情况;DHE荧光探针观察肝细胞活性氧(ROS)水平;酶标仪检测肝组织亚铁离子含量及脂质过氧化水平(MDA);Western blotting法及免疫组化染色检测肝组织GPX4表达情况。结果 清胰汤中、高剂量均可以明显改善小鼠肝缺血再灌注损伤,有效降低小鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)水平(P<0.05),减少肝组织缺血坏死情况,降低肝细胞ROS水平,并且能够有效降低肝组织亚铁离子含量及脂质过氧化水平(P<0.05),恢复GPX4活性(P<0.05)。结论 清胰汤中、高剂量均可减轻模型小鼠的肝脏氧化应激,减少肝缺血坏死,改善肝功能,对于肝缺血再灌注损伤的肝脏具有保护作用,其潜在机制可能与干预肝铁死亡的发生有关,且其作用呈剂量依赖性。Objective:To observe the protective effect and mechanism of Qingyi Decoction(QYD)on hepatic ischemia-reperfusion injury in mice.Methods:A total of 30 male SPF C57BL/6 mice were randomly divided into 5 groups:sham,model,low-dose QYD,mid-dose QYD and high-dose QYD,with 6 mice each.Mice in QYD groups were given corresponding doses of drugs continuously 7 days before modeling,while mice in sham and model groups were given same saline.The model of 70%hepatic ischemia-reperfusion injury was established by clamping the arteriovenous blood flow of left lobe and middle lobe of liver with 60 min ischemia and 12 h reperfusion.The following items were detected:liver enzymes by ELISA,hepatic ischemia performance by HE staining,hepatocytes′reactive oxygen species(ROS)by DHE fluorescent probe,level of ferrous ion and lipid peroxidation by enzyme-labeling measuring instrument,GPX4 expression by Western blotting and immunohistochemical staining.Results:Mid-and high-dose QYD could significantly improve hepatic ischemia-reperfusion injury in mice,including effectively reducing the level of serum ALT and AST(P<0.05),the liver ischemia performance,ROS in hepatocytes,and effectively decreasing the content of ferrous ions and lipid peroxidation in liver(P<0.05).It also restored the activity of GPX4(P<0.05)as well.Conclusion:Both mid-and high-dose QYD could reduce liver oxidative stress level,hepatic ischemia performance and improve liver function.It proves that QYD has the protective effect on hepatic ischemia-reperfusion injury and its potential mechanism might be related to the reduction of liver ferroptosis with a dose-dependent manner.

关 键 词：脓毒症相关肝损伤 肝缺血再灌注损伤 清胰汤 铁死亡 小鼠 

分 类 号：R285.5[医药卫生—中药学]