四烯甲萘醌(MK-4)对CCl4诱导的急性肝损伤小鼠模型的保护作用分析  

Menaquinone-4 exerts a protective effect against carbon tetrachloride-induced acute liver injury in mice by alleviating ferroptosis

作　　者：叶露 赵凡 黄倩倩 张佳怡 王建青[1,2,3] YE Lu;ZHAO Fan;HUANG Qianqian;ZHANG Jiayi;WANG Jianqing(Department of Pharmacy,Anhui Medical University,Hefei 230032,China;Department of Pharmacy,The First Affiliated Hospital of Anhui Medical University,Hefei 230012,China;Anhui Public Health Clinical Center,Hefei 230012,China)

机构地区：[1]安徽医科大学药学院,合肥230032 [2]安徽医科大学第一附属医院药学部,合肥230012 [3]安徽省公共卫生临床中心,合肥230012

出　　处：《临床肝胆病杂志》2024年第1期121-128,共7页Journal of Clinical Hepatology

基　　金：国家自然科学基金(82073566);安徽省高校优秀青年人才支持计划资助项目(gxyq2019014);临床药学与药理学共建项目(2020);安徽省公共卫生临床中心安徽医科大学第一附属医院北区科研培育基金资助项目(2023YKJ14,2023YKJ06,2023YKJ11)。

摘　　要：目的 探究四烯甲萘醌(MK-4)通过减轻铁死亡对四氯化碳(CCl_(4))诱导的急性肝损伤(ALI)小鼠的保护作用及其机制。方法 选用8周龄成年ICR雄鼠,适应性饲养后,分为Control组、MK-4组、CCl_(4)模型组(6、12、24 h)、MK-4+CCl_(4)组(6、12、24 h),每组6只。Control组腹腔注射同等剂量玉米油;MK-4组腹腔注射40 mg/kg的MK-4溶液,1 h后腹腔注射同等剂量玉米油;MK-4+CCl_(4)组(6、12、24 h)先腹腔注射40 mg/kg的MK-4溶液,1 h后和CCl_(4)模型组(6、12、24 h)同时腹腔注射0.3 mL/kg CCl_(4)溶液,分别在6、12、24 h进行取材。通过HE染色观察小鼠肝脏的病理变化;普鲁士蓝染色检测肝组织中铁聚集;生化仪检测小鼠血清中AST和ALT水平;试剂盒检测肝匀浆中组织铁含量、氧化应激指标丙二醛(MDA)、谷胱甘肽(GSH)水平;RT-PCR检测铁死亡标志基因长链脂酰辅酶A合成酶4(ACSL4)、前列腺素内过氧化物酶2(PTGS2)和谷胱甘肽过氧化物酶4(GPX4)以及铁代谢相关基因铁调素调节蛋白(HJV)、转铁蛋白受体1(TFR1)和膜转铁蛋白(FPN)的表达,Western Blot检测GPX4的蛋白表达水平。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 在时效研究中,与Control组相比,CCl_(4)模型组(12、24 h)的肝质量系数和血清ALT、AST水平明显上升(P值均<0.05),且通过HE染色结果也反映出,肝损伤程度随着时间延长逐渐加重。同时,MK-4+CCl_(4)(6、12、24 h)组和CCl_(4)模型组(6、12、24 h)相比时,MK-4+CCl_(4)(12 h)组的肝质量系数和血清ALT、AST水平下降(P值均<0.05),且肝组织的坏死面积减少,因此在后续研究中采用12 h的小鼠各组织样本进行检测。与Control组相比,CCl_(4)组MDA上升,GSH下降,差异均有统计学意义(P值均<0.05);与CCl_(4)组相比,MK-4+CCl_(4)组MDA下降,GSH上升,差异均有统计学意义(P值均<0.05)。铁死亡关键指标ASCL4、PTGS2,与Control组相比,CCl_(4)组显著上升,GPX4则显著下�Objective To investigate whether menaquinone-4(MK-4)can exert a protective effect against carbon tetrachloride(CCl_(4))-induced acute liver injury(ALI)in mice by alleviating ferroptosis.Methods After adaptive feeding,adult male ICR mice,aged 8 weeks,were divided into Control group,MK-4 group,CCl_(4) model group(6-hour,12-hour,and 24-hour),and MK-4+CCl_(4) group(6-hour,12-hour,and 24-hour),with 6 mice in each group.The mice in the Control group were given intraperitoneal injection of an equal dose of corn oil;the mice in the MK-4 group were given intraperitoneal injection of 40 mg/kg MK-4 solution,followed by an equal dose of corn oil after 1 hour;the mice in the MK-4+CCl_(4) group(6-hour,12-hour,and 24-hour)were given intraperitoneal injection of 40 mg/kg MK-4 solution,and after 1 hour,the mice in this group and the CCl_(4) model group(6-hour,12-hour,and 24-hour)were given intraperitoneal injection of 0.3 mL/kg CCl_(4) solution,with samples collected at 6,12,and 24 hours.HE staining was used to observe the pathological changes of mouse liver;Prussian blue staining was used to observe iron accumulation in liver tissue;a biochemical analyzer was used to measure the serum levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT);related kits were used to measure the levels of tissue iron content and the oxidative stress indices malondialdehyde(MDA)and glutathione(GSH)in liver homogenate;RT-PCR was used to measure the expression levels of ferroptosis marker genes(acyl-CoA synthetase long-chain family member 4[ACSL4],prostaglandin-endoperoxide synthase 2[PTGS2],and glutathione peroxidase 4[GPX4])and iron metabolism-related genes(hemojuvelin[HJV],transferrin receptor 1[TFR1],and ferroportin[FPN]),and Western blot was used to measure the protein expression level of GPX4.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results In the aging study,compared with the

关 键 词：肝功能衰竭 急性 铁死亡 甲萘醌4 小鼠 近交ICR 

分 类 号：R575[医药卫生—消化系统] R-332[医药卫生—内科学]