莱菔硫烷通过激活Keap1/Nrf2信号通路减轻敌草快诱导的小鼠急性肝损伤  

Sulforaphane alleviates acute liver injury induced by diquat in mice by activating Keap1/Nrf2 signaling pathway

作　　者：王建红 彭亮[1] 吴潦章 黄山 何国丽 沈沛 梁婧 黄婷婷 黄家明 钟红 周满红 Wang Jianhong;Peng Liang;Wu Liaozhang;Huang Shan;He Guoli;Shen Pei;Liang Jing;Huang Tingting;Huang Jiaming;Zhong Hong;Zhou Manhong(Department of Emergency,Guizhou Aerospace Hospital,Zunyi 563003,Guizhou,China;Department of Emergency,Affiliated Hospital of Zunyi Medical University,Zunyi 563003,Guizhou,China;Department of Emergency,Kweichow Moutai Hospital,Zunyi 564500,Guizhou,China)

机构地区：[1]贵州航天医院急诊科,贵州遵义563003 [2]遵义医科大学附属医院急诊科,贵州遵义563003 [3]贵州茅台医院急诊科,贵州遵义564500

出　　处：《中华危重病急救医学》2024年第11期1183-1189,共7页Chinese Critical Care Medicine

基　　金：国家自然科学基金(82060346);贵州省卫生健康委科技基金(gzwkj2021-007);贵州省遵义市科技计划项目(2018-131)。

摘　　要：目的探讨莱菔硫烷(SFN)对敌草快(DQ)中毒致小鼠急性肝损伤的保护作用及其可能机制。方法将48只雄性C57BL/6小鼠按随机数字表法分成对照组(Control组)、DQ模型组(DQ组)、SFN干预组(DQ+SFN组)、SFN对照组(SFN组),每组12只。通过一次性腹膜腔注射40 mg/kg DQ溶液1 mL建立急性肝损伤小鼠模型;SFN组注射1 mL的ddH2O。制模后4 h,DQ+SFN组和SFN组腹膜腔注射5 mg/kg SFN溶液0.5 mL,每日1次,连续7 d;DQ组和Control组注射等量ddH2O。干预7 d后处死小鼠收集肝脏组织、血液样本,检测血浆中生物标志物丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平,以及肝组织中氧化应激指标超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)、丙二醛(MDA)水平;透射电镜下观察肝组织结构改变;荧光显微镜下观察肝组织中细胞凋亡情况和活性氧(ROS)水平;蛋白质免疫印迹试验(Western blotting)检测肝组织中核转录因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)、Kelch样ECH相关蛋白1(Keap1)、活化的天冬氨酸特异性半胱氨酸蛋白酶9(cleaved caspase-9)的蛋白表达。结果与Control组比较,DQ组肝脏线粒体严重肿胀,基质部分溶解,大量嵴断裂缺失;血浆AST和ALT水平显著升高;肝脏MDA含量升高,SOD、GSH活性下降,ROS水平显著升高,凋亡细胞明显增多,Nrf2、HO-1蛋白表达明显下降,Keap1、cleaved caspase-9蛋白表达明显升高。与DQ组比较,DQ+SFN组线粒体损伤减轻,血浆AST和ALT水平明显下降〔ALT(U/L):58.22±4.39比79.94±3.32,AST(U/L):177.64±8.40比219.62±11.60,均P<0.01〕,肝脏MDA含量明显下降,SOD、GSH活性明显升高〔MDA(μmol/g):5.63±0.18比5.96±0.29,SOD(kU/g):102.05±4.01比84.34±5.34,GSH(mmol/g):16.32±1.40比13.12±1.84,均P<0.05〕,肝组织中ROS产生明显减少〔ROS(荧光强度):115.90±10.89比190.70±10.16,P<0.05〕,凋亡细胞明显减少(细胞凋亡指数:4.39±1.00比10.71±0.56,P<0.01),Nrf2、HO-1蛋白表达明显升高,Keap1、cleaved caspasObjective To investigate the protective effect and possible mechanism of sulforaphane(SFN)on acute liver injury in mice induced by diquat(DQ)poisoning.Methods Forty-eight male C57BL/6 mice were divided into Control group,DQ model group(DQ group),SFN intervention group(DQ+SFN group),and SFN control group(SFN group)using a random number table method,with 12 mice in each group.Acute liver injury mice model was established by one-time intraperitoneal injection of 1 mL of 40 mg/kg DQ solution at once.SFN group was injected with 1 mL of ddH 2O.After 4 hours of molding,0.5 mL of 5 mg/kg SFN solution was injected into the peritoneal cavity of the DQ+SFN group and SFN group,once daily for 7 consecutive days.DQ group and Control group were injected with an equal amount of ddH 2O.Then,the mice were euthanized to collect liver tissue and blood samples,and the levels of plasma biomarkers alanine aminotransferase(ALT)and aspartate aminotransferase(AST),as well as oxidative stress indicators such as superoxide dismutase(SOD),glutathione(GSH),and malondialdehyde(MDA)in liver tissue were measured.The changes of liver structure were observed under transmission electron microscopy.The apoptosis and reactive oxygen species(ROS)level in liver tissue were observed under fluorescence microscope.Western blotting was used to detect the protein expressions of nuclear factor E2-related factor 2(Nrf2),hemeoxygenase-1(HO-1),Kelch-like ECH-associated protein 1(Keap1),and cleaved caspase-9 in liver tissue.Results Compared with the Control group,the liver mitochondria in the DQ group showed severe swelling,partial dissolution of the matrix,and cristae rupture and loss;the levels of plasma AST and ALT significantly increased,the MDA content in the liver increased,the activities of SOD and GSH decreased,the level of ROS significantly increased,the number of apoptotic cells in the liver significantly increased,the protein expressions of Nrf2 and HO-1 significantly decreased,and the protein expressions of Keap1 and cleaved caspase-9 significantly i

关 键 词：莱菔硫烷 敌草快 核转录因子E2相关因子2 抗氧化反应元件 活性氧 

分 类 号：R595.4[医药卫生—内科学]