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目的 探讨乙型肝炎病毒e抗原(hepatitis B e antigen,HBeAg)对Bewo细胞维甲酸诱导基因-I(retinoic acidinducible-Ⅰ,RIG-Ⅰ)和线粒体抗病毒信号蛋白(mitochondrial antiviral signaling protein,MAVS)表达的作用.方法 首先以终浓度2 μg/mL的RIG-Ⅰ配体poly(dAT∶dAT)处理Bewo细胞24 h,观察RIG-Ⅰ、MAVS mRNA的表达.然后将5μgHBeAg重组质粒pcDNA3.1 (+)-HBe转染Bewo细胞,转染48 h后,以2μg/mL的poly(dAT∶ dAT)刺激24 h.采用实时荧光定量反转录PCR和酶联免疫吸附测定分别检测细胞RIG-Ⅰ、MAVS mRNA表达及上清干扰素-β(interferon-β,IFN-β)水平.并采用免疫荧光染色法观察HBeAg对细胞核因子-κB (nuclear factor-κB,NF-κB)p65入核的影响.结果 poly(dAT∶ dAT)可诱导Bewo细胞RIG-Ⅰ、MAVS mRNA表达(27.86±9.49,21.75±8.04),高于对照组(14.51±6.27,11.59±4.73)(P＜0.01).转染5μg HBeAg重组质粒组poly(dAT∶ dAT)可诱导Bewo细胞RIG-Ⅰ、MAVS mRNA表达(13.33±4.23,10.48±3.17)及IFN-β产生[(14.20±5.01)pg/mL],低于对照组[27.86±9.49,21.75 ±8.04;(47.69± 14.3)pg/mL](P＜0.01),但与pcDNA3.1-HBe-5组[11.82±3.58,9.39±3.22;(10.82± 3.75)pg/mL]比较,差异无统计学意义(P＞0.05).且HBeAg可干扰poly(dAT∶dAT)诱导Bewo细胞NF-κB p65入核,抑制NF-κB的激活.结论 HBeAg可下调Bewo细胞RIG-Ⅰ、MAVS mRNA的表达,并抑制NF-κB入核及IFN-β产生,这可能是乙型肝炎病毒(hepatitis B virus,HBV)逃避宿主免疫应答的一种重要策略.“,”Objective To explore the effect of hepatitis B e antigen(HBeAg) on the expression of retinoic acidinducible-Ⅰ(RIG-Ⅰ) and mitochondrial antiviral signaling protein(MAVS) in Bewo cells.Methods Firstly,Bewo ceils were exposed to RIG-Ⅰ ligand poly(dAT∶ dAT) with final concentration of 2 μg/mL for 24 h to observe the expression of RIG-Ⅰ,MAVS mRNA.Secondly,5 μg HBeAg recombinant vector pcDNA3.1(+)-HBe was transfected into Bewo cells,after 48 h,the cells were treated with 2 μg/mL poly(dAT∶ dAT) for 24 h.To assay RIG-Ⅰ,MAVS mRNA expression in Bewo cells and interferon-β(IFN-β) level in supernatant by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay,respectively.And to observe the effect of HBeAg on poly(dAT∶ dAT)-induced NF-κB p65 nuclear translocaton by immunofluorescent staining.Results Poly(dAT:dAT) could induce RIG-Ⅰ,MAVS mRNA expression(27.86± 9.49,21.75 ± 8.04) in Bewo cells,significantly higher than the control group (14.51 ±6.27,11.59±4.73)(P ＜ 0.01).Poly(dAT∶ dAT)-induced RIG-Ⅰ,MAVS mRNA expression (13.33±4.23,10.48±3.17) and IFN-β level [(14.20±5.01)pg/mL] in transfecting 5 μg HBeAg recombinant vector group were strikingly lower the control group [27.86±9.49,21.75±8.04,(47.69± 14.3)pg/mL] (P ＜ 0.01),but no significant difference with the pcDNA3.1-HBe-5 group [11.82±3.58,9.39±3.22,(10.82±3.75)pg/mL](P ＞ 0.05).And HBeAg could disturb NF-κB p65 nuclear translocaton induced by poly(dAT∶dAT),the activity of NF-κB was inhibited significantly.Conclusions HBeAg could downregulate the expression of RIG-Ⅰ,MAVS mRNA,and inhibit NF-κB p65 nuclear translocaton and IFN-β levels in Bewo cells,which was an important strategy for HBV avoiding host immune responses.