姜黄素可通过减轻线粒体氧化应激改善NF-κB/NLRP3介导的ARDS肾损伤  

Curcumin alleviates nuclear factor-κB/NOD-like receptor protein 3 mediated renal injury caused by acute respiratory distress syndrome through reducing mitochondrial oxidative stress

作　　者：杨茂宪 田海涛[2] 沈鹏 徐龙生 刘鹤[4] 朱建刚 王倩倩 施云超 Yang Maoxian;Tian Haitao;Shen Peng;Xu Longsheng;Liu He;Zhu Jian'gang;Wang Qianqian;Shi Yunchao(Department of Critical Care Medicine,First Hospital of Jiaxing(Affiliated Hospital of Jiaxing College),Jiaxing 314001,Zhejiang,China;Department of Anesthesiology,First People's Hospital of Jining,Jining 266021,Shandong,China;Department of Central Laboratory,First Hospital of Jiaxing(Affiliated Hospital of Jiaxing College),Jiaxing 314001,Zhejiang,China;Department of Anesthesiology,Huzhou Central Hospital,Huzhou 313003,Zhejiang,China)

机构地区：[1]嘉兴市第一医院(嘉兴学院附属医院)重症医学科,浙江嘉兴314001 [2]济宁市第一人民医院麻醉科,山东济宁266021 [3]嘉兴市第一医院(嘉兴学院附属医院)中心实验室,浙江嘉兴314001 [4]湖州市中心医院麻醉科,浙江湖州313003

出　　处：《中华危重病急救医学》2023年第4期393-397,共5页Chinese Critical Care Medicine

基　　金：国家自然科学基金(82171227);浙江省嘉兴市医学重点学科建设计划项目(2019-ZC-12);浙江省嘉兴市科技计划项目(2018AY32017);浙江省嘉兴市医学领军人才培养人选(2019-lj-03)。

摘　　要：目的评价姜黄素对急性呼吸窘迫综合征(ARDS)大鼠肾组织线粒体氧化应激、核转录因子-κB/NOD样受体蛋白3(NF-κB/NLRP3)炎症小体信号通路及组织细胞损伤的影响。方法将24只SPF级健康雄性SD大鼠按照随机数字表法分为对照组、ARDS模型组及姜黄素小剂量和大剂量组,每组6只。气管内给予脂多糖(LPS)4 mg/kg雾化吸入制备ARDS大鼠模型;对照组给予生理盐水2 mL/kg。姜黄素小剂量和大剂量组于制模后24 h分别灌胃姜黄素100 mg/kg或200 mg/kg,每日1次;对照组和ARDS模型组给予等量生理盐水。7 d后处死大鼠取下腔静脉血,采用酶联免疫吸附试验(ELISA)检测血清中性粒细胞明胶酶相关脂质运载蛋白(NGAL)水平。取肾组织,采用ELISA法检测活性氧(ROS)水平;采用黄嘌呤氧化酶法检测超氧化物歧化酶(SOD)活性;采用比色法检测丙二醛(MDA)水平;采用蛋白质免疫印迹试验(Western blotting)检测低氧诱导因子-1α(HIF-1α)、天冬氨酸特异性半胱氨酸蛋白酶3(caspase-3)、NF-κB p65、Toll样受体4(TLR4)的蛋白表达;采用反转录-聚合酶链反应(RT-PCR)检测HIF-1α、NLRP3、白细胞介素-1β(IL-1β)的mRNA表达;采用原位末端缺刻标记法(TUNEL)检测肾组织细胞凋亡;透射电镜下观察肾小管上皮细胞及线粒体形态变化。结果与对照组比较,ARDS模型组大鼠表现为肾脏氧化应激和炎症反应,肾损伤标志物NGAL血清含量明显升高,NF-κB/NLRP3炎症小体信号通路被激活,肾组织细胞凋亡率明显升高,且透射电镜下可见肾小管上皮细胞损伤,线粒体完整性破坏,提示模型成功诱发肾损伤。给予姜黄素干预后,大鼠肾小管上皮细胞及线粒体损伤均明显减轻,同时氧化应激明显减轻,NF-κB/NLRP3炎症小体信号通路被抑制,肾组织细胞凋亡率明显降低,并均呈一定剂量依赖性;与ARDS模型组比较,姜黄素大剂量组血清NGAL和肾组织MDA、ROS水平均明显降低〔NGAL(μg/L):13.8±1.7�Objective To evaluate the effect of curcumin on renal mitochondrial oxidative stress,nuclear factor-κB/NOD-like receptor protein 3(NF-κB/NLRP3)inflammatory body signaling pathway and tissue cell injury in rats with acute respiratory distress syndrome(ARDS).Methods A total of 24 specific pathogen free(SPF)-grade healthy male Sprague-Dawley(SD)rats were randomly divided into control group,ARDS model group,and low-dose and high-dose curcumin groups,with 6 rats in each group.The ARDS rat model was reproduced by intratracheal administration of lipopolysaccharide(LPS)at 4 mg/kg via aerosol inhalation.The control group was given 2 mL/kg of normal saline.The low-dose and high-dose curcumin groups were administered 100 mg/kg or 200 mg/kg curcumin by gavage 24 hours after model reproduction,once a day.The control group and ARDS model group were given an equivalent amount of normal saline.After 7 days,blood samples were collected from the inferior vena cava,and the levels of neutrophil gelatinase-associated lipocalin(NGAL)in serum were determined by enzyme-linked immunosorbent assay(ELISA).The rats were sacrificed,and kidney tissues were collected.Reactive oxygen species(ROS)levels were determined by ELISA,superoxide dismutase(SOD)activity was detected using the xanthine oxidase method,and malondialdehyde(MDA)levels were determined by colorimetric method.The protein expressions of hypoxia-inducible factor-1α(HIF-1α),caspase-3,NF-κB p65,and Toll-like receptor 4(TLR4)were detected by Western blotting.The mRNA expressions of HIF-1α,NLRP3,and interleukin-1β(IL-1β)were detected by reverse transcription-polymerase chain reaction(RT-PCR).Renal cell apoptosis was detected by TdT-mediated dUTP nick end labeling(TUNEL).The morphological changes in renal tubular epithelial cells and mitochondria were observed under a transmission electron microscope.Results Compared with the control group,the ARDS model group exhibited kidney oxidative stress and inflammatory response,significantly elevated serum levels of kidney injury biomar

关 键 词：急性呼吸窘迫综合征 肾脏 线粒体 NLRP3炎症小体 氧化应激 

分 类 号：R285.5[医药卫生—中药学]