辅酶Q10抑制IL-17/NF-κB蛋白表达对敌草快中毒大鼠肾损伤保护作用  

Protective effect of coenzyme Q10 on renal injury in diquat poisoned rats by inhibiting the expression of IL-17/NF-κB protein

作　　者：杨登会 吴瑾 胡杰 詹江珊 陆安静 赖福平 贾映茂 陆元兰 Yang Denghui;Wu Jin;Hu Jie;Zhan Jiangshan;Lu Anjing;Lai Fuping;Jia Yingmao;Lu Yuanlan(Emergency Department of Affiliated Hospital of Zunyi Medical University,Zunyi 563003,China;Outpatient Department of Affiliated Hospital of Zunyi Medical University,Zunyi 563003,China)

机构地区：[1]遵义医科大学附属医院急诊科,遵义563003 [2]遵义医科大学附属医院门诊部,遵义563003

出　　处：《中华急诊医学杂志》2024年第9期1249-1256,共8页Chinese Journal of Emergency Medicine

基　　金：国家自然科学基金(82060245);贵州省科技计划项目(黔科合基础-ZK[2022]一般655);贵州省卫健委科技基金(gzwkj2022-284);遵义市科技计划项目(遵市科合HZ字(2023)217号。

摘　　要：目的探究抗氧化剂辅酶Q10(coenzyme Q10,CoQ10)是否通过抗氧化应激作用抑制白介素(interleukin,IL)-17及核转录因子-κB(nuclear factor kappa-B,NF-κB)信号通路参与调控敌草快中毒(diquat,DQ)所致大鼠的肾损伤,该机制是否与减轻线粒体功能障碍相关。方法体内及体外模型中检测核因子κB抑制蛋白α、磷酸化核转录因子κB(nuclear factor kappa B,P-NF-κB)、JNK相关亮氨酸拉链蛋白(JNK-related leucine zipper protein,JLP)、神经保护蛋白PTEN诱导的假定激酶1(neuroprotective protein PTEN-induced putative kinase 1,PINK1)通路蛋白的表达;生化检测肾损伤标志物及炎症细胞因子:血清尿素氮(serum urea nitrogen,BUN)、血肌酐(Creatinine,Cr)、血清胱抑素-C(Cystatin C,CysC)、肾损伤分子1、丙二醛、超氧化物歧化酶(supemxide dismutase,SOD)、中性粒细胞明胶酶相关脂质运载蛋白(neutrophilgelatinase-Associated lipocalin,NGAL)等;肾脏病理HE染色观察光镜下肾损伤程度及病理评分;免疫荧光检测活性氧表达情况;CCK-8实验观察给药后细胞增殖水平。结果体内实验DQ组染毒72 h后血浆及肾脏标本中肾功能损伤指标(Cr、BUN、CysC、NGAL、KIM-1)明显增高,存在明显的病理组织学改变、病理评分升高;离体实验HK-2细胞DQ染毒48 h后,细胞活力下降一半。DQ染毒后氧化应激指标血清SOD下降、丙二醛上升,肾组织活性氧免疫荧光值上升,予以CoQ10干预后能减轻DQ诱导的大鼠病理损害及增强HK-2细胞活力,减轻肾损伤,减轻氧化应激水平。此外,体内实验DQ组染毒后促炎细胞因子(IL-6、TNF-α和IL-17)水平表达增加,体内DQ组和体外HK-2细胞DQ组检测P-NF-κBp65蛋白表达量明显升高,线粒体功能障碍指标PINK1蛋白表达量明显升高,JLP蛋白、IκB-α蛋白表达量明显下降;予以CoQ10干预后,能减低P-NF-κBp65蛋白、PINK1的表达,增加IκB-α蛋白的表达及减轻JLP降解,证明CoQ10可改善DQ中毒后线粒体功能障碍�Objective To explore whether antioxidant coenzyme Q10(CoQ10)is involved in the regulation of renal injury induced by diquat poisoning(DQ)in rats through anti-oxidative stress and inhibition of interleukin(IL)-17 and nuclear factor kappa-B(NF-κB)signaling pathway,and whether this mechanism is related to alleviating mitochondrial dysfunction.Methods The expressions of NFκB inhibitory proteinα(IKB-α),phosphorylated nuclear factorκB(P-NF-κB),JNK-related leucine zipper protein(JLP)and neuroprotective protein PTEN-induced putative kinase 1(PINK1)pathway proteins were detected in vivo and in vitro.Biochemical detection of renal injury markers and inflammatory cytokines:serum urea nitrogen(BUN),serum creatinine(Cr),Cystatin C(CysC),renal injury molecule 1,Malondialdehyde,Supemxidedismutase(SOD),neutrophil gelatinase-associated lipocalin(NGAL),etc.Renal pathology HE staining was used to observe the degree of renal injury and pathological score under light microscope.The expression of reactive oxygen species(ROS)was detected by immunofluorescence.CCK-8 experiment was used to observe the level of cell proliferation after administration.Results In vivo experiment,the indexes of renal function injury(Cr,BUN,CysC,NAGL,KIM-1)in plasma and kidney samples were significantly increased after 72 h of exposure in DQ group,and there were significant histopathological changes and pathological scores increased.In vitro experiment HK-2 cells were exposed to DQ for 48 h,and the cell viability decreased by half.After exposure to DQ,serum SOD decreased,MDA increased,and the immunofluorescence value of ROS in renal tissue increased.Intervention with CoQ10 can alleviate the pathological damage induced by DQ in rats,enhance the vitality of HK-2 cells,alleviate renal injury and reduce the level of oxidative stress.In addition,the expression of proinflammatory cytokines(IL-6,TNF-αand IL-17)increased in DQ group in vivo,the expression of P-NFκBp65 protein in DQ group in vivo and HK-2 cell DQ group in vitro increased significantly,the expr

关 键 词：敌草快中毒 肾损害 氧化应激 辅酶Q10 NF-ΚB 

分 类 号：R595[医药卫生—内科学]