核因子E2相关因子2蛋白通过调控铁死亡介导脓毒症相关性肝损伤  

Nuclear factor E2-related factor 2 protein mediates sepsis-associated liver injury by regulating ferroptosis

作　　者：钟朕 欧阳训彦 郭起财 熊力[1] 刘先发[1] Zhong Zhen;Ouyang Xunyan;Guo Qicai;Xiong Li;Liu Xianfa(Department of Emergency,First Affiliated Hospital of Gannan Medical University,Ganzhou 341000,Jiangxi,China;Department of Orthopedics,First Affiliated Hospital of Gannan Medical University,Ganzhou 341000,Jiangxi,China)

机构地区：[1]赣南医学院第一附属医院急诊科,江西赣州341000 [2]赣南医学院第一附属医院骨科,江西赣州341000

出　　处：《中华危重病急救医学》2024年第5期491-495,共5页Chinese Critical Care Medicine

基　　金：江西省卫生健康委科技计划项目(202310802)。

摘　　要：目的探讨核因子E2相关因子2(Nrf2)蛋白对小鼠脓毒症相关性肝损伤(SALI)铁死亡的作用。方法按随机数字表法将雄性SD小鼠分成6组,每组6只。采用盲肠结扎穿孔术(CLP)构建小鼠SALI模型,假手术(Sham)组仅开腹处理。CLP+Fer-1组、CLP+Erastin组、CLP+ML385组、CLP+Curcumin组分别于术后经腹腔注射铁死亡抑制剂Ferrostatin-1(Fer-1)10 mg·kg^(-1)·d^(-1)、铁死亡激活剂Erastin 20 mg·kg^(-1)·d^(-1)、Nrf2抑制剂ML38530 mg·kg^(-1)·d^(-1)、Nrf2激活剂Curcumin 100 mg·kg^(-1)·d^(-1)进行干预;Sham组、CLP组予生理盐水10 mg·kg^(-1)·d^(-1),均连续给药10 d。术后10 d收集小鼠血清和肝组织,检测血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST),以及肝组织匀浆液中丙二醛(MDA)、谷胱甘肽(GSH)和Fe2+水平;苏木素-伊红(HE)染色后光镜下观察肝组织病理学改变;透射电镜下观察肝细胞线粒体形状和长度;蛋白质免疫印迹试验(Western blotting)检测肝组织Nrf2、谷胱甘肽过氧化物酶4(GPX4)、前列腺素-内过氧化物合酶2(PTGS2)的蛋白表达。结果与Sham组比较,CLP组小鼠血清ALT、AST水平明显升高;镜下可见肝索排列紊乱,细胞肿胀、坏死,线粒体长度明显变短;肝组织中MDA、Fe2+水平明显上升,GSH含量明显下降,Nrf2、GPX4蛋白表达下降,PTGS2蛋白表达明显升高。与CLP组比较,CLP+Fer-1组和CLP+Curcumin组血清ALT、AST水平明显降低〔ALT(U/L):80.65±19.44、103.45±20.52比283.50±37.12,AST(U/L):103.33±11.90、127.33±15.79比288.67±36.82,均P<0.05〕;镜下可见肝索欠规整,细胞轻微肿胀,线粒体长度明显增加(μm:1.42±0.09、1.43±0.21比1.07±0.25,均P<0.05);肝组织中MDA、Fe2+水平明显下降,GSH含量明显升高〔MDA(mol/g):0.87±0.23、1.85±0.43比4.47±0.95,Fe2+(μg/g):63.80±7.15、67.48±6.28比134.52±14.32,GSH(mol/g):1.95±0.29、1.95±0.45比0.55±0.29,均P<0.05〕;肝组织Nrf2、GPX4蛋白表达明显升高,PTGS2蛋白表达明显下降(Nrf2/GAPDH:1.80±Objective To investigate the effect of nuclear factor E2-related factor 2(Nrf2)protein on ferroptosis in mice with sepsis-associated liver injury(SALI).Methods The male Sprague-Dawley(SD)mice were divided into 6 groups according to the random number table method,with 6 mice in each group.The SALI model of mice was established by cecal ligation and puncture(CLP),and the Sham group was only treated with laparotomy.CLP+Fer-1 group,CLP+Erastin group,CLP+ML385 group and CLP+Curcumin group were intraperitoneally injected with iron death inhibitor Ferrostatin-1(Fer-1)10 mg·kg^(-1)·d^(-1),iron death activator Erastin 20 mg·kg^(-1)·d^(-1),Nrf2 inhibitor ML38530 mg·kg^(-1)·d^(-1)and Nrf2 activator Curcumin 100 mg·kg^(-1)·d^(-1)after CLP,respectively;Sham group and CLP group were given normal saline 10 mg·kg^(-1)·d^(-1),each group was administered continuously for 10 days.Ten days after operation,the serum and liver tissues of mice were collected to detect the levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in serum,and the levels of malondialdehyde(MDA),glutathione(GSH)and Fe2+in liver homogenate.The pathological changes of liver tissue were observed under light microscope after hematoxylin-eosin(HE)staining.The shape and length of mitochondria in liver cells were observed under transmission electron microscope.The protein expressions of Nrf2,glutathione peroxidase 4(GPX4)and prostaglandin-endoperoxide synthase 2(PTGS2)in liver tissue were detected by Western blotting.Results Compared with Sham group,the serum levels of ALT and AST in the CLP group were significantly increased;histologically,the hepatic cord was disordered,the cells were swollen and necrotic,and the length of mitochondria was significantly shortened;the levels of MDA and Fe2+in liver tissue increased significantly,and the content of GSH decreased significantly;the protein expressions of Nrf2 and GPX4 in liver tissue decreased,and the protein expression of PTGS2 increased significantly.Compared with CLP group,the serum le

关 键 词：脓毒症相关性肝损伤 铁死亡 核因子E2相关因子2 

分 类 号：R459.7[医药卫生—急诊医学] R575[医药卫生—治疗学]