基于NF-κB/NLRP3/Caspase-1信号通路探究大黄素对草酸钙肾结石模型大鼠的影响  

Effects of emodin on calcium oxalate kidney stone model rats through NF-κB/NLRP3/Caspase-1 signaling pathway and mechanism study

作　　者：李瑞杰 项瑞君 郑树眺 刘柏良 周浩 杨旭凯 LI Ruijie;XIANG Ruijun;ZHENG Shutiao;LIU Baiiang;ZHOU hao;YANG Xukai(Department of Comprehensive Surgery,940th Hospital of Joint Logistic Support Force of Chinese People’s Liberation Army,Lanzhou 730050,Gansu Province,China;First School of Clinical Medical,Gansu University of Chinese Medicince,Lanzhou 730000,Gansu Province,China;Gansu Provincial Key Laboratory of Stem Cell and Gene Drug,Lanzhou 730050,Gansu Province,China)

机构地区：[1]联勤保障部队第940医院综合外科,甘肃兰州730050 [2]甘肃中医药大学第一临床医学院,甘肃兰州730000 [3]甘肃省干细胞与基因药物重点实验室,甘肃兰州730050

出　　处：《解放军医学院学报》2024年第11期1159-1166,共8页Academic Journal of Chinese PLA Medical School

基　　金：甘肃省自然科学基金(21JR11RA008);兰州市人才创新创业项目(2021-RC-106)。

摘　　要：背景肾结石作为泌尿系统常见疾病,发病率和复发率逐年升高,目前早期治疗肾盐结晶的有效药物依旧较少。目的基于NF-κB/NLRP3/Caspase-1信号通路初步探究大黄素对草酸钙(calcium oxalate,CaOx)肾结石大鼠模型结石形成和肾功能的影响,以及相关作用机制。方法将40只SPF级Wistar雄性大鼠随机分为4组,即对照组、模型组、干预组和阳性对照组,每组10只。除对照组外,其余各组饮用0.75%乙二醇溶液,隔日2%氯化铵溶液灌胃诱导CaOx肾结石大鼠模型,同时干预组和阳性对照组分别灌胃大黄素和肾石通颗粒,连续给药4周后,检测尿液中草酸盐、枸橼酸、Ca^(2+)、Mg^(2+)水平以及血清肾损伤标志物;HE染色和Vonkossa染色分别观察大鼠肾组织病理变化和钙盐沉积;酶联免疫吸附法(ELISA)测定肾组织中氧化应激损伤标志物;采用实时荧光定量聚合酶链式反应(Real-time PCR)法检测肾组织中NLRP3、IL-1β、IL-18 mRNA水平,蛋白免疫印迹法(Western blot)检测肾组织NLRP3、NF-κB p56、Caspase-1、IL-1β、IL-18蛋白水平。结果与对照组相比,模型组大鼠体质量、尿Mg^(2+)、枸橼酸下降(P<0.01),肾指数、尿Ca2+、尿草酸盐、血清肌酐、尿素氮、尿酸升高(P<0.01);肾组织可见明显病理损伤和钙盐沉积;肾组织中丙二醛(malondialdehyde,MDA)、活性氧(reactive oxygen species,ROS)含量升高,NLRP3、IL-1β、IL-18 mRNA水平以及NLRP3、IL-1β、IL-18、NF-κB p56、Caspase-1蛋白水平升高,超氧化物歧化酶(superoxide dismutase,SOD)含量降低(P<0.01)。与模型组相比,大黄素组和肾石通颗粒组大鼠体质量、尿Mg^(2+)、枸橼酸升高(P<0.01),肾指数、尿Ca^(2+)、尿草酸盐、血清肌酐、尿素氮、尿酸降低(P<0.01);肾组织病理损伤和钙盐沉积明显减轻;肾组织中MDA、ROS含量降低,NLRP3、IL-1β、IL-18 mRNA水平以及NLRP3、IL-1β、IL-18、NF-κB p56、Caspase-1蛋白水平降低,SOD水平升高(P<0.01)。结论大黄素Background As a common disease of the urinary system,the incidence and recurrence of kidney stones are increasing year by year,and there are still few effective drugs for the early treatment of renal salt crystals.Objective To preliminarily investigate the effects and possible mechanisms of action of rhodopsin on renal function in calcium oxalate(CaOx)kidney stone model rats based on the NF-κB/NLRP3/Caspase-1 signaling pathway.Methods Forty SPF-grade Wistar male rats were randomly divided into control group,model group,intervention group,positive control group,with 10 rats in each group.The CaOx kidney stone rat model was induced by drinking 0.75%ethylene glycol solution and gavage with 2%ammonium chloride solution every other day in all groups except the control group,while the intervention group and the positive control group were gavaged with emodin and shenshitong granules,respectively.After 4 weeks of continuous drug administration,urine levels of oxalate,citrate,Ca^(2+),and Mg^(2+)were measured;serum markers of renal impairment and serum markers of renal injury were measured;HE staining and Vonkossa staining were used to observe the histopathological changes and calcium salt deposition in rat kidney,respectively;Enzyme-linked immunosorbent assay(ELISA)for the determination of markers of oxidative stress injury in renal tissues;NLRP3,IL-1β,and IL-18 mRNA levels in kidney tissues were measured by real-time fluorescence quantitative polymerase chain reaction(real-time PCR).Protein immunoblotting(Western-blotting)was used to detect kidney tissue NLRP3,NF-κB p56,Caspase-1,IL-1β,and IL-18 protein levels.Results Compared with the control group,body mass,urinary Mg^(2+),and citrate decreased(P<0.01),and renal index,urinary Ca^(2+),and urinary oxalate with serum creatinine,urea nitrogen,and uric acid increased(P<0.01)in rats of the model group;Significant pathological damage and calcium salt deposits could be seen in the renal tissue;MDA and ROS content and NLRP3,IL-1β,and IL-18 mRNA levels and NLRP3,IL-1β,IL-

关 键 词：草酸钙肾结石 大黄素 NF-κB/NLRP3/Caspase-1信号通路 作用机制 治疗 

分 类 号：R692.4[医药卫生—泌尿科学]