右美托咪定预处理通过调控p62/Keap1/Nrf2途径减轻脂多糖诱导的大鼠肝脏氧化应激、炎症和铁死亡  

Dexmedetomidine pretreatment alleviates lipopolysaccharide(LPS)-induced hepatic oxidative stress,inflammation and ferroptosis in rats via the p62/Keap1/Nrf2 signaling pathway

作　　者：沈耘[1] 吴佳艺 韩艳艳 钱淑雯[1] 郭旋[1] SHEN Yun;WU Jiayi;HAN Yanyan;QIAN Shuwen;GUO Xuan(Department of Anesthesiology,Yangpu Central Hospital,Shanghai 200000,China)

机构地区：[1]上海市杨浦区中心医院麻醉科,上海200000

出　　处：《安徽医药》2025年第3期472-481,I0001,共11页Anhui Medical and Pharmaceutical Journal

基　　金：上海市卫生系统优秀青年医学人才培养项目(PWRq2022-27)。

摘　　要：目的研究右美托咪定(Dex)对脂多糖(LPS)诱导的肝脏氧化应激、炎症及铁死亡的影响及其可能的机制。方法2022年8月至2023年6月,腹腔注射LPS制备大鼠肝损伤模型。50只雄性Wistar大鼠按随机数字表法分为五组:对照组、Dex组、LPS组、LPS+Dex组和LPS+Dex+p62活化抑制剂(XRK3F2)组,每组各10只。Kaplan‒Meier法进行生存分析;酶联免疫吸附测定(ELISA)大鼠血清中天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)、肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-1β和IL-6含量;苏木素-伊红(HE)染色观察大鼠肝组织的形态学变化;流式细胞术检测肝脏组织中活性氧簇含量;丙二醛、谷胱甘肽和总谷胱甘肽(T-GSH)含量,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性及谷胱甘肽/氧化型谷胱甘肽(GSH/GSSG)比值采用试剂盒进行检测;实时荧光定量PCR(RT-qPCR)检测炎症因子、铁死亡及信号通路相关分子mRNA的表达;大鼠肝组织中Fe^(2+)含量用组织铁测定试剂盒检测;透射电镜观察大鼠肝组织细胞中线粒体形态;蛋白质印迹法检测铁死亡及信号通路相关分子蛋白的表达。结果与对照组相比,LPS组大鼠生存率和体质量明显降低,肝组织湿/干比及AST和ALT酶活力显著增加,肝组织形态发生损坏、炎症和坏死细胞增加;相较于LPS组,LPS+Dex组大鼠生存率和体质量明显增加,肝水肿、肝功能受损及肝组织病理学损伤明显减轻。与对照组相比,LPS可导致大鼠肝组织中活性氧簇和丙二醛含量增加,SOD、CAT和GSH-Px酶活性降低,谷胱甘肽和T-GSH含量和GSH/GSSG比值减少;Dex预处理则减弱了LPS诱导的氧化应激参数的改变。与对照组相比,LPS组大鼠血清中TNF-α、IL-1β和IL-6含量和肝组织中TNF-α、IL-1β和IL-6 mRNA表达升高;Dex则降低了LPS诱导的炎症因子的表达和释放。与对照组相比,LPS组大鼠肝组织细胞线粒体皱缩、嵴减少、膜密度增加,Objective To explore the effects of dexmedetomidine(Dex)on lipopolysaccharide(LPS)-induced liver oxidative stress,inflammation,and ferroptosis and its underlying mechanisms.Methods The study was conducted from August 2022 to June 2023,and the rat model of liver injury was established by intraperitoneal injection of LPS.A total of 50 male Wistar rats were divided into 5 groups:control group,Dex group,LPS group,LPS+Dex group,and LPS+Dex+p62 activation inhibitor(XRK3F2)group,each group consisting of 10 rats.Kaplan‒Meier was performed to analyze survival rate.Enzyme-linked immunosorbent assays(ELISAs)were conducted to measure aspartate transaminase(AST),alanine aminotransferase(ALT),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)content in sera of the rats.Hematoxylin-eosin(HE)staining was used to observe the morphological changes of liver tissue.Reactive oxygen species(ROS)production in the liver was detected by flow cytometry.Malondialdehyde,glutathione and total glutathione(T-GSH)content,superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GSH-Px)activities,as well as glutathione/reduced glutathione(GSH/GSSG)ratio were measured by commercial kits.The mRNA expressions of inflammatory mediators,ferroptosis-related molecules,and signaling pathway-associated molecules were assessed by RT-qPCR analyses.Fe^(2+)content in the liver was measured by tissue iron assay kit.Transmission electron microscopy was used to observe the mitochondrial morphology.Western blot was conducted to detect the protein levels of ferroptosis-and signal pathway-related molecular proteins in the liver tissues.Results Compared with control group,LPS significantly decreased survival rate and body weight but increased the liver wet/dry ratio and activities of AST and ALT.LPS also led to severe liver damage as shown by increased inflammatory and necrotic cells.Compared with LPS group,Dex preconditioning raised survival rate and body weight,alleviated liver edema,liver function impairment and liver histo

关 键 词：肝损伤 右美托咪定 氧化应激 炎症 铁死亡 p62蛋白/Kelch样ECH2相关蛋白1/核因子E2相关因子2通路 

分 类 号：R614[医药卫生—麻醉学]