乙肝病毒截短型表面抗原中蛋白与X蛋白对肾小管上皮细胞核转录因子的影响  

Role of NF-κB activiation by MHBs^(t167)/HBx in human renal tubular cells

作　　者：洪柳[1,2] 张静[1] 李青[1] 闵婕[1] 陆建荣[1] 李烦繁[1] 李航[1] 郭双平[1] 

机构地区：[1]第四军医大学西京医院病理科,西安710032 [2]海军总医院病理科,北京100048

出　　处：《肾脏病与透析肾移植杂志》2010年第2期135-141,共7页Chinese Journal of Nephrology,Dialysis & Transplantation

基　　金：国家自然科学基金(30400213)

摘　　要：目的:目的研究167例乙肝病毒截短型表面抗原中蛋白(C-terminally truncated middle size surface proteins,MHBst167)和X蛋白(hepatitis B virus X protein,HBx)对肾小管上皮细胞核转录因子κB(nuclear factor-kappaB,NF-κB)活化的影响及其相关机制探讨。方法:肾小管上皮细胞系(HK-2)转染mhbst167或(和)hbx后,蛋白印迹法检测NF-κB核易位及其抑制蛋白(inhibitor of nuclear factor-kappa B,IκBα)磷酸化水平的变化,凝胶电泳迁移率和双萤光素酶报告基因分析进一步检测NF-κB活性;通过对蛋白激酶C(protein kinase C,PKC)活性和细胞外调节激酶(extracellular regulated protein kinases,ERK)磷酸化水平检测探讨NF-κB活化的机制。结果:HK-2细胞转染mhbst167或(和)hbx基因后,NF-κB核易位、磷酸化IκBα、κB结合活性及κB基因转录均增加(P<0.05);且PKC激酶活性和磷酸化ERK水平也增加(P<0.05),而Raf蛋白均无表达。结论:在肾小管上皮细胞中,MHBst167/HBx可能通过PKC/ERK通路(非Raf依赖性)活化NF-κB。Objective：To investigate the effect of MHBs^t167/HBx on NF-κB activation in human renal tubular cells and its possible mechanism. Methodology ： After renal tubular cells （ HK-2 ） being transfected with the genes encoding MHBst167 and/or HBx, nuclear translocation of nuclear factor-kappa B （NF-κB） and inhibitor of nuclear factor-kappa B （IκBα） phosphorylation were assessed by Western blot. Electrophoretic mobility shift assays （EMSA） and dual luciferase reporter assays （DLR） were used to further examine NF-κB activation following transfection. The possible mechanisms for NF-κB activation were analyzed by protein kinase C （PKC） activity assay and Western blot for phosphor-ERK. Results： Cells transfected with mhbs^t167 and/or hbx increased NF-κB nuclear translocation, phosphor-IκBα, KB-DNA binding activity and κB-dependent transcription in compared with the control （ P 〈 0. 05 ）. Moreover, the PKC activity and phosphor-ERK were also increased （P 〈 0. 05 ） during the NF-κB activation process, but the Raf-1 protein in all groups was negative. Condusion：MHBs^t167/HBx maybe induced NF-κB activation via the PKC/ERK pathway （ independent of Raf pathway） in human renal tubular cells.

关 键 词：肾小管上皮细胞截短型表面抗原中蛋白X蛋白 核转录因子-κB蛋白激酶C/c—Raf/细胞外调节激酶 

分 类 号：R692.3[医药卫生—泌尿科学]