TRAF6抑制剂C25-140对敌草快中毒致小鼠急性肾损伤的保护作用  

Protective effect of tumor necrosis factor receptor-associated factor 6 inhibitor C25-140 on acute kidney injury induced by diquat poisoning in mice

作　　者：黄婷婷 饶国胜 赵志杰 徐娜娜 周满红 欧仁阳 Huang Tingting;Rao Guosheng;Zhao Zhijie;Xu Nana;Zhou Manhong;Ou Renyang(Department of Emergency,Affiliated Hospital of Zunyi Medical University,Zunyi 563003,Guizhou,China;Department of Emergency,Kweichow Moutai Hospital,Renhuai 564500,Guizhou,China)

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

出　　处：《中华危重病急救医学》2024年第12期1273-1278,共6页Chinese Critical Care Medicine

基　　金：国家自然科学基金(82060346);贵州省研究生教育创新计划项目(YJSCXJH2018-092)。

摘　　要：目的探讨肿瘤坏死因子受体相关因子6(TRAF6)抑制剂C25-140对急性敌草快(DQ)中毒致小鼠急性肾损伤(AKI)的保护作用及机制。方法将80只SPF级健康雄性C57BL/6小鼠随机分为正常对照组、DQ模型组、C25-140干预组和C25-140对照组,每组20只。采用一次性腹膜腔注射40 mg/kg DQ溶液1 mL的方法建立DQ中毒小鼠模型;正常对照组和C25-140对照组腹膜腔注射等量纯水。制模4 h后,C25-140干预组和C25-140对照组腹膜腔注射C25-1405 mg/kg;DQ模型组和正常对照组腹膜腔注射等量纯水;每日1次,持续7 d。7 d后麻醉小鼠取眼球血,处死后收集肾组织。光镜下观察肾组织病理变化;透射电镜下观察肾组织超微结构及线粒体改变;部分血清送检血肌酐(SCr)及血尿素氮(BUN)水平;酶联免疫吸附试验(ELISA)测定血清白细胞介素(IL-6、IL-1β)和肿瘤坏死因子-α(TNF-α)水平;蛋白质免疫印迹试验(Western blotting)检测肾组织TRAF6、髓样分化因子88(MyD88)、核转录因子-κB(NF-κB)蛋白表达水平;化学法测定血清丙二醛(MDA)和超氧化物歧化酶(SOD)的含量。结果正常对照组小鼠观察期间行为无异常;DQ模型组小鼠在染毒后逐渐出现精神倦怠、皮毛蓬松、活动行为减少、进食少等情况,严重者死亡;C25-140干预组小鼠上述表现较DQ模型组减轻。光镜下显示,DQ模型组HE染色可见炎症细胞浸润,肾小球硬化及近端小管扩张、空泡化;而C25-140干预组炎症反应较DQ模型组减轻。透射电镜下显示,DQ模型组线粒体损伤程度相对较高,严重肿胀,体积增大,基质溶解,嵴断裂缺失;C25-140干预组线粒体损伤程度较DQ模型组减轻。与正常对照组比较,DQ模型组血清SCr、BUN、IL-6、IL-1β、TNF-α、MDA水平均明显升高,血清SOD水平明显降低;与DQ模型组比较,C25-140干预组血清SCr、BUN、IL-6、IL-1β、TNF-α、MDA水平均明显降低〔SCr(μmol/L):59.07±13.11比83.61±20.13,BUN(mmol/L):25.83±9.95比40.78±Objective To investigate the protective effect and mechanism of tumor necrosis factor receptor-associated factor 6(TRAF6)inhibitor C25-140 on acute kidney injury(AKI)induced by acute diquat(DQ)poisoning in mice.Methods A total of 80 SPF grade healthy male C57BL/6 mice were randomly divided into the normal control group,DQ model group,C25-140 intervention group,and C25-140 control group,with 20 mice in each group.The DQ poisoning mouse model was established by using one-time intraperitoneal injection of 1 mL of 40 mg/kg DQ solution.The normal control group and C25-140 control group were injected with an equal amount of pure water into the peritoneal cavity.After 4 hours of model establishment,the C25-140 intervention group and C25-140 control group were given intraperitoneal injection of C25-1405 mg/kg.The normal control group and DQ model group were given equal amounts of pure water,once a day for 7 consecutive days.After 7 days,the mice were anesthetized,eye blood was collected,and renal tissue was collected after sacrifice.The pathological changes of renal tissue were observed under a light microscope and renal tissue structure and mitochondrial changes were observed under transmission electron microscopy.The levels of serum creatinine(SCr)and blood urea nitrogen(BUN)were measured.Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of serum interleukins(IL-6,IL-1β)and tumor necrosis factor-α(TNF-α).Western blotting was used to detect the protein expression levels of TRAF6,myeloid differentiation factor 88(MyD88),and nuclear factor-κB(NF-κB)in renal tissue.Chemical method was used to determine the content of serum malondialdehyde(MDA)and superoxide dismutase(SOD).Results During the observation period,there were no abnormal behaviors in the normal control group mice.The DQ model group mice gradually showed symptoms such as mental fatigue,fluffy fur,reduced activity,and low food intake after being exposed to the toxin,and severe cases resulted in death.The above symptoms were alleviated in

关 键 词：敌草快 肿瘤坏死因子受体相关因子6 急性肾损伤 氧化应激 

分 类 号：R595.4[医药卫生—内科学] R692[医药卫生—临床医学]