OBJECTIVE Hepatitis B virus X protein (HBx) is a pivotal factor for HBV-induced hepatitis. Herein, we sought to investigate HBx-mediated NLR pyrin domain containing 3 (NLRP3) inflammasome activation and pyroptosis under oxidative stress. METHODS The effect of HBx on the NLRP3 inflammasome was analyzed by enzyme-linked immunosorbent assays, quantitative reverse transcription-polymerase chain reaction, western blotting, and immunofluorescence in hepatic HL7702 cells. Pyroptosis was evaluated by western blotting, lactate dehydrogenase release, propidium iodide staining, and transmission electron microscopy. NLRP3 expression in the inflammasome from liver tissues was assessed by immunohistochemistry. RESULTS In hydrogen peroxide (H2O2)-stimulated HL7702 cells, HBx triggered the release of pro-inflammatory mediators apoptosis-associated speck-like protein containing a CARD (ASC), interleukin (IL)-1β, IL-18, and high-mobility group box 1 (HMGB1); activated NLRP3; and initiated pro-inflammatory cell death (pyroptosis). HBx localized to the mitochondria, where it induced mitochondrial damage and production of mitochondrial reactive oxygen species (mitoROS). Treatment of HL7702 cells with a mitoROS scavenger attenuated HBx-induced NLRP3 activation and pyroptosis. Expression levels of NLRP3, ASC, and IL-1β in liver tissues from patients were positively correlated with HBV DNA concentration. CONCLUSIONS The NLRP3 inflammasome was activated by elevated mitoROS levels and mediated HBx-induced liver inflammation and hepatocellular pyroptosis under H2O2-stress conditions.

中文翻译：

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乙型肝炎病毒X蛋白通过NLRP3炎症小体激活，在氧化应激下促进肝细胞凋亡。

目的乙型肝炎病毒X蛋白（HBx）是HBV引起的肝炎的关键因素。在本文中，我们试图研究在氧化应激下含有3（NLRP3）炎性小体活化和焦磷酸化的HBx介导的NLR吡啶结构域。方法通过酶联免疫吸附试验，定量逆转录-聚合酶链反应，蛋白质印迹和肝HL7702细胞免疫荧光分析HBx对NLRP3炎性小体的影响。通过蛋白质印迹，乳酸脱氢酶释放，碘化丙啶染色和透射电子显微镜评估细胞凋亡。通过免疫组织化学评估了来自肝脏组织的炎性小体中的NLRP3表达。结果在过氧化氢（H2O2）刺激的HL7702细胞中，HBx触发了促炎介质与凋亡相关的斑点样蛋白的释放，该蛋白包含CARD（ASC），白介素（IL）-1β，IL-18和高迁移率族框1（HMGB1）；激活的NLRP3; 并引发促炎性细胞死亡（化脓）。HBx定位于线粒体，它在其中诱导线粒体损伤并产生线粒体活性氧（mitoROS）。用mitoROS清除剂处理HL7702细胞会减弱HBx诱导的NLRP3激活和凋亡。患者肝组织中NLRP3，ASC和IL-1β的表达水平与HBV DNA浓度呈正相关。结论在H2O2胁迫条件下，高水平的mitoROS水平激活了NLRP3炎性体，并介导了HBx诱导的肝脏炎症和肝细胞凋亡。白介素（IL）-1β，IL-18和高迁移率分组盒1（HMGB1）；激活的NLRP3; 并引发促炎性细胞死亡（化脓）。HBx定位于线粒体，它在其中诱导线粒体损伤并产生线粒体活性氧（mitoROS）。用mitoROS清除剂处理HL7702细胞会减弱HBx诱导的NLRP3激活和凋亡。患者肝组织中NLRP3，ASC和IL-1β的表达水平与HBV DNA浓度呈正相关。结论NLRP3炎性体在高H2O2胁迫条件下被高水平的mitoROS激活并介导HBx诱导的肝脏炎症和肝细胞凋亡。白介素（IL）-1β，IL-18和高迁移率分组盒1（HMGB1）；激活的NLRP3; 并引发促炎性细胞死亡（化脓）。HBx定位于线粒体，它在其中诱导线粒体损伤并产生线粒体活性氧（mitoROS）。用mitoROS清除剂处理HL7702细胞会减弱HBx诱导的NLRP3激活和凋亡。患者肝组织中NLRP3，ASC和IL-1β的表达水平与HBV DNA浓度呈正相关。结论在H2O2胁迫条件下，高水平的mitoROS水平激活了NLRP3炎性体，并介导了HBx诱导的肝脏炎症和肝细胞凋亡。它会诱导线粒体损伤并产生线粒体活性氧（mitoROS）。用mitoROS清除剂处理HL7702细胞会减弱HBx诱导的NLRP3激活和凋亡。患者肝组织中NLRP3，ASC和IL-1β的表达水平与HBV DNA浓度呈正相关。结论在H2O2胁迫条件下，高水平的mitoROS水平激活了NLRP3炎性体，并介导了HBx诱导的肝脏炎症和肝细胞凋亡。它诱导线粒体损伤并产生线粒体活性氧（mitoROS）。用mitoROS清除剂处理HL7702细胞会减弱HBx诱导的NLRP3激活和凋亡。患者肝组织中NLRP3，ASC和IL-1β的表达水平与HBV DNA浓度呈正相关。结论在H2O2胁迫条件下，高水平的mitoROS水平激活了NLRP3炎性体，并介导了HBx诱导的肝脏炎症和肝细胞凋亡。患者肝组织中IL-1β和IL-1β与HBV DNA浓度呈正相关。结论在H2O2胁迫条件下，高水平的mitoROS水平激活了NLRP3炎性体，并介导了HBx诱导的肝脏炎症和肝细胞凋亡。患者肝组织中IL-1β和IL-1β与HBV DNA浓度呈正相关。结论在H2O2胁迫条件下，高水平的mitoROS水平激活了NLRP3炎性体，并介导了HBx诱导的肝脏炎症和肝细胞凋亡。