乙型肝炎病毒X蛋白对小鼠足细胞补体调节蛋白CD59和Crry表达的影响  

Hepatitis B virus X protein upregulates the expression of CD59 and Crry in mouse podocytes

作　　者：尹晓玲[1] 周建华[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院儿,科武汉430030

出　　处：《中华儿科杂志》2010年第12期934-938,共5页Chinese Journal of Pediatrics

基　　金：基金项目：国家自然科学基金面上项目（30772360）

摘　　要：目的 乙型肝炎病毒相关性肾炎（HBV-GN）所引起的膜性肾病与原发膜性肾病有所区别,其肾组织中攻膜复合物（C5b-9）阳性率显著低于原发膜性肾病,说明在乙型肝炎病毒相关性膜性肾病（HBV-MN）中补体的活动受到某种抑制,产生这种差异的机制尚不清楚.本研究旨在探讨乙型肝炎病毒X蛋白（HBx）是否影响足细胞补体调节蛋白CD59和Crry表达,从而抑制补体活化.方法 培养的小鼠足细胞分为腺病毒载体乙型肝炎病毒X基因（Ad-HBx）转染组（Ad-X组）、空白足细胞组（B组）和腺病毒空载体转染组（Ad组）,P38、PI-3K、ERK1/2通路抑制剂（SB203580,LY294002,U0126）及二甲基亚砜（DMSO）干预组分别标记为SB组、LY组、U0126组和DMSO组.RT-PCR和流式细胞术检测Ad-HBx转染对CD59和Crry基因和蛋白的表达,MTT法检测其对补体激活的影响 观察P38、PI-3K、ERK1/2通路抑制剂及DMSO对足细胞CD59和Crry表达的影响.结果 B组、Ad组和Ad-X组CD59表达率分别为（17.71±3.81）%、（18.29±3.36）%和（45.7±9.01）% B组、Ad组和Ad-X组Crry蛋白表达率分别为（18.00±2.31）%、（21.78±2.01）%和（47.45±9.95）%,Ad-X组CD59和Crry蛋白表达率显著高于B组和Ad组（P值均＜0.005） Ad-X组CD59和Crry mRNA光密度明显强于B组和Ad组 SB组、LY组、U0126组和DMSO组CD59表达率分别为（17.35±1.24）%,（46.19±9.77）%,（43.03±6.83）%和（40.04±6.39）% Crry蛋白表达率分别为（18.14±3.56）%,（31.95±1.68）%,（31.95±1.69）%和（37.14±3.92）%,与Ad-X组比较,SB组CD59和Crry蛋白表达率明显降低（P值均＜0.05）,MTT法提示在各个血清稀释点上,Ad-X组细胞裂解抑制率均明显高于B组和Ad组（P值均＜0.05）.结论 HBx可能通过活化P38MAPK通路上调小鼠足细胞CD59和Crry蛋白和基因的表达,抑制补体介导的足细胞裂解,这可能导致HBV在足细胞潜伏感染,不易被清除,从而在HBV-GN发生中发挥作用.Objective Different from primary membranous nephropathy, hepatitis B virus associated membranous nephropathy （HBV-MN） shows lower deposits of membrane attack complex （C5b-9）in glomerular subepithelium. The causes of relatively low complement activation in this disease remain unclear. The aim of this study was to investigate the influence of hepatitis B x protein （HBx） on the expression of CD59 and Crry in mouse podocytes. Method Cultured mouse podocytes were divided into adenovirus vector hepatitis B virus X gene （Ad-HBx） transfected group （Ad-X group）, blank podocytes group （B group） and adenovirus vector transfected group （Ad group）. CD59 and Crry mRNA expression were assayed by semiquantitative RT-PCR. CD59 and Crry expression were tested by flow cytometry. The effect of HBx on complement activation was evaluated with MTT method. And then, the effects of P38MAPK, PI-3K and ERK1/2 pathway inhibitors （ SB203580, LY294002, U0126） and DMSO on CD59 and Crry expression were respectively detected by flow cytometry. Result Proteins CD59 and Crry expression rates （ % ） in group B, Ad group and Ad-X group were 17. 71 ± 3. 81, 18. 29 ± 3.36 and 45.7 ±9. 01 18 ±2. 31,21.78 ±2. 01 and 47.45 ±9.95, respectively. Compared with group B, CD59 and Crry expression in group Ad was not significantly different （ P values for both 〉 0. 05 ）, but CD59 and Crry protein expression in Ad-X group was significantly higher than that in groups B and Ad （ P values for both 〈0. 005 ） CD59 and Crry gene expression in group Ad was not significantly different from that in group B （P values for both 〉 0. 05 ）. However, CD59 and Crry gene expression of Ad-X group was significantly higher than that in groups B and Ad （P values for both 〈 0. 05 ）. Flow cytometry detected CD59 protein expression rates （ % ） were 17. 35 ± 1.24, 46. 19 ± 9. 77, 43. 03 ± 6. 83 and 40. 04 ± 6. 39 and Crry protein expression rates （ % ） were 18. 14 ± 3. 56, 31.95 ± 1.68, 31.95 ± 1.69 and 37

关 键 词：反式激活因子类 抗原 CD59 受体 补体 

分 类 号：R512.62[医药卫生—内科学]