泛素调控蛋白A20对慢性乙肝患者单核细胞活性的影响  

Influence of ubiquitin-editing protein A20 to monocytes activity in chronic hepatitis B patients

作　　者：王佳斌[1] 李英兰[1] 张希功 马燕春 WANG Jiabin;LI Yinglan;ZHANG Xigong;MA Yanchun(Department of General Medicine,the People's Hospital of Qinghai Province,Xining 810007,China)

机构地区：[1]青海省人民医院全科医学科,西宁810007

出　　处：《中国免疫学杂志》2022年第20期2528-2533,共6页Chinese Journal of Immunology

基　　金：青海省基础研究计划项目(2020-ZJ-931)资助。

摘　　要：目的:观察泛素调控蛋白A20在慢性乙肝患者外周血中的变化及其对CD14^(+)单核细胞功能的影响。方法:选取慢性乙肝患者47例和健康对照者26例,纯化外周血CD14^(+)单核细胞和CD4^(+)T细胞,流式细胞术检测CD14^(+)单核细胞中A20水平,实时定量PCR检测CD14^(+)单核细胞中A20 mRNA相对表达量。A20 siRNA或对照siRNA转染慢性乙肝患者纯化的CD14^(+)单核细胞,ELISA检测上清中炎症细胞因子水平,实时定量PCR检测FasL和TRAIL mRNA相对表达量。CD14^(+)单核细胞与HepG2.2.15细胞共培养,检测CD14^(+)单核细胞诱导靶细胞死亡的比例。CD14^(+)单核细胞与CD4^(+)T细胞共培养,流式细胞术检测CD4^(+)T细胞分泌IFN-γ和IL-17的比例。结果:慢性乙肝患者外周血CD14^(+)单核细胞中A20^(+)细胞比例高于健康对照者(P=0.0004),慢性乙肝患者外周血CD14^(+)单核细胞中A20平均荧光强度和A20 mRNA相对表达量亦高于健康对照者(P<0.001)。A20 siRNA转染慢性乙型肝炎患者纯化的CD14^(+)单核细胞后,分泌炎症细胞因子水平高于未转染细胞和对照siRNA转染细胞(P<0.05),但FasL和TRAIL mRNA未见明显变化(P>0.05)。A20 siRNA转染的CD14^(+)单核细胞诱导靶细胞死亡的比例高于未转染细胞和对照siRNA转染细胞(P<0.05),诱导CD4^(+)T细胞分泌IFN-γ和IL-17的比例亦高于未转染细胞和对照siRNA转染细胞(P<0.05)。结论:慢性乙肝患者CD14^(+)单核细胞中高表达的A20有抑制细胞毒性及CD4^(+)T细胞活化的作用,可能与导致乙型肝炎病毒感染慢性化相关。Objective:To investigate the change of ubiquitin-editing protein A20 and its influence on CD14^(+)monocytes function in chronic hepatitis B patients.Methods:Forty-seven chronic hepatitis B patients and twenty-six healthy controls were enrolled.Peripheral CD14^(+)monocytes and CD4^(+)T cells were purified.A20 level in CD14^(+)monocytes was assessed by flow cytometry,while A20 mRNA relative level was semi-quantified by real-time PCR.Purified CD14^(+)monocytes from chronic hepatitis B patients were transfected by A20 siRNA or control siRNA.Levels of proinflammatory cytokines in cultured supernatants were measured by ELLISA.FasL and TRAIL mRNA relative levels were semi-quantified by real-time PCR.CD14^(+)monocytes were co-cultured with HepG2.2.15cells,and percentage of target cell death was investigated.CD14^(+)monocytes were also co-cultured with CD4^(+)T cells,and percentage of IFN-γand IL-17-secreting CD4^(+)T cells were assessed by flow cytometry.Results:Percentage of A20-positive cells in CD14^(+)monocytes was elevated in chronic hepatitis B patients when compared with healthy controls(P=0.0004).Mean fluorescence intensity and mRNA relative level of A20 in CD14^(+)monocytes was also increased in chronic hepatitis B patients when compared with healthy controls(P<0.001).Proinflammatory cytokine levels were increasingly secreted by chronic hepatitis B patients derived CD14^(+)monocytes with A20 siRNA transfection when compared with untransfection and control siRNA transfection(P<0.05),however,there were no significant differences of FasL or TRAIL mRNA relative level(P>0.05).Percentage of target cell death induced by CD14^(+)monocytes was elevated in A20 siRNA transfected cells when compared with untransfected and control siRNA transfected cells(P<0.05),percentage of IFN-γ-secreting and IL-17-secreting CD4^(+)T cells,which induced by CD14^(+)monocytes were also increased when compared with untransfected and control siRNA transfected cells(P<0.05).Conclusion:Increased level of A20 in CD14^(+)monocytes from chronic

关 键 词：病毒性肝炎 乙型 慢性 泛素调控蛋白A20 单核细胞 

分 类 号：R392.12[医药卫生—免疫学]