泼尼松对阿霉素肾病大鼠肾组织FAK、RANK、RANKL、p38MAPK、NF-κB表达的影响  

The effects of prednisone on the expressions of FAK,RANK,RANKL,p38MAPK and NF-κB in renal tissues of rats with adriamycin-induced nephropathy

作　　者：陈小英[1] 董晓登 陈虹[1] 赖晶晶 严焱 郑京[1] CHEN Xiaoying

机构地区：[1]福建中医药大学附属人民医院,福建福州350004 [2]福建中医药大学,福建福州350122

出　　处：《中医临床研究》2022年第5期6-10,共5页Clinical Journal Of Chinese Medicine

基　　金：福建省中青年教师教育科研项目(JAT200180);福建省卫健委创新课题(2017CX41)。

摘　　要：目的:观察泼尼松对阿霉素肾病大鼠粘着斑激酶(Focal Adhesion Kinase,FAK)、核因子-κB受体活化因子(Receptor Activator of Nuclear Factor Kappa B,RANK)、核因子-κB受体活化因子配体(Receptor Activator of Nuclear Factor Kappa B Ligand,RANKL)、p38丝裂原活化蛋白激酶(p38 Mitogen-activated Protein Kinase,p38MAPK)、核转录因子-κB(Nuclear Factor-κB,NF-κB)表达的影响。方法:利用随机数字表法把SD大鼠分为正常组(尾静脉一次性注射等量生理盐水)、模型组(注射盐酸阿霉素6.5 mg/kg造模)、治疗组(成功造模后以泼尼松每天10 mg/kg灌胃),在实验不同的时间点测定24 h尿蛋白定量,观察肾组织电镜结果,用电泳迁移率变动分析(Electrophoretic Mobility Shift Assay,EMSA)检测NF-κB含量,用实时荧光定量聚合酶链式反应(Quantitative Real-time Fluorescence Polymerase Chain Reaction,qRT-PCR)检测FAK、p38MAPK、RANK、RANKL、Nephrin表达水平的变化,用Western blot检测肾组织FAK、RANK、RANKL、Nephrin蛋白以及p38MAPK蛋白和磷酸化蛋白。结果:①与正常组相比,模型组尿蛋白水平升高(P<0.01);用药后的治疗组尿蛋白水平下降(P<0.01);②模型组电镜下可见足细胞足突广泛融合、肿胀或增宽,治疗组明显减轻;③模型组PCR结果示FAK、p38MAPK、RANKL、RANK mRNA表达较正常组升高(P<0.01),治疗组FAK、p38MAPK、RANKL、RANK mRNA表达较模型组降低(P<0.01);④与正常组相比,模型组RANK、RANKL蛋白以及FAK、p38MAPK总蛋白和磷酸化表达升高(P<0.01),Nephrin蛋白表达减少(P<0.01);与模型组相比,治疗组RANK、RANKL蛋白以及FAK、p38MAPK总蛋白和磷酸化表达降低(P<0.05),Nephrin蛋白表达升高(P<0.05);⑤EMSA检测结果显示,与正常组相比,模型组NF-κB表达增多(P<0.01);与模型组相比,治疗组NF-κB表达水平降低(P<0.01)。结论:阿霉素肾病肾组织中有RANK/RANKL的表达,经激素治疗,FAK-p38MAPK/RANKL-RANK-p38MAPK的通路被抑制,使NF-κB入核减少,炎症反Objective:To observe the effects of prednisone on the expressions of FAK,RANK,RANKL,p38MAPK,NF-κB in rats with adriamycin-induced nephropathy.Methods:Using the random number table method,SD rats were divided into the normal group(took equal amount of saline injection through the tail vein),the model group(took doxorubicin hydrochloride injection of 6.5 mg/kg for modeling),and the treatment group(received intragastric prednisone of 10 mg/kg daily after successful modeling).24 h urine protein quantification was measured at different time points in the experiment.The renal tissue electron microscopy results were observed.The NF-κB content was detected by EMSA.The FAK,p38MAPK,RANK,RANKL and Nephrin levels were detected by quantitative real-time fluorescent PCR.Western blot was used to detect renal tissue FAK,RANK,RANKL,Nephrin protein,p38MAPK protein and phosphorylated protein.Results:①Compared with the normal group,the urine protein level in the model group was increased(P<0.01).The urine protein level in the treatment group was decreased after medication(P<0.01).②Under the electron microscope,it can be seen that the foot processes of the podocytes in the model group were extensively fused,swollen or widened,and the foot processes of the podocytes in the treatment group were significantly reduced.③The qRT-PCR results in the model group showed that the expressions of FAK,p38MAPK,RANKL,RANK mRNA were higher than the normal group(P<0.01),and the expressions of FAK,p38MAPK,RANKL,RANK mRNA in the treatment group was lower than the model group(P<0.01).④Compared with the normal group,the expressions of RANK,RANKL protein,FAK,p38MAPK total protein and phosphorylation in the model group were increased(P<0.01),and the expression of Nephrin protein was decreased(P<0.01).Compared with the model group,the expressions of RANK,RANKL protein,FAK,and p38MAPK total protein and phosphorylation in the treatment group were decreased(P<0.05),and the expression of Nephrin protein in the treatment group was increased(P<0.05).⑤T

关 键 词：阿霉素肾病 核转录因子-κB 核因子-κB受体活化因子/核因子-κB受体活化因子配体 泼尼松 P38丝裂原活化蛋白激酶 

分 类 号：R692[医药卫生—泌尿科学]