Hepatitis B Virus X Protein Promotes Liver Cell Pyroptosis Under Oxidative Stress Through NLRP3 Inflammasome Activation

Overview

Journal Inflamm Res

Specialties Allergy & Immunology
Pathology

Date 2020 Apr 30

PMID 32347316

Citations 54

Authors

Wen-Hui Xie

Jian Ding

Xiao-Xia Xie

Xiao-Huang Yang

Xiao-Fan Wu

Zhi-Xin Chen

Qi-Lan Guo

Wen-Yu Gao

Xiao-Zhong Wang

Dan Li

Affiliations

Soon will be listed here.

Abstract

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 (HO)-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 HO-stress conditions.

Citing Articles

Hepatic Iron Overload and Hepatocellular Carcinoma: New Insights into Pathophysiological Mechanisms and Therapeutic Approaches.

Chatzikalil E, Arvanitakis K, Kalopitas G, Florentin M, Germanidis G, Koufakis T Cancers (Basel). 2025; 17(3).

PMID: 39941760 PMC: 11815926. DOI: 10.3390/cancers17030392.

Dual role of pyroptosis in liver diseases: mechanisms, implications, and therapeutic perspectives.

Yang S, Zou Y, Zhong C, Zhou Z, Peng X, Tang C Front Cell Dev Biol. 2025; 13:1522206.

PMID: 39917567 PMC: 11798966. DOI: 10.3389/fcell.2025.1522206.

Endothelial Dysfunction and Liver Cirrhosis: Unraveling of a Complex Relationship.

Nesci A, Ruggieri V, Manilla V, Spinelli I, Santoro L, Di Giorgio A Int J Mol Sci. 2024; 25(23).

PMID: 39684569 PMC: 11640898. DOI: 10.3390/ijms252312859.

Emerging Roles of High-mobility Group Box-1 in Liver Disease.

Wang L, Dong Z, Zhang Y, Peng L J Clin Transl Hepatol. 2024; 12(12):1043-1056.

PMID: 39649031 PMC: 11622203. DOI: 10.14218/JCTH.2024.00317.

Unveiling the nexus: pyroptosis and its crucial implications in liver diseases.

Miao Z, Zhang X, Xu Y, Liu Y, Yang Q Mol Cell Biochem. 2024; .

PMID: 39477911 DOI: 10.1007/s11010-024-05147-1.

References

Wree A, Eguchi A, McGeough M, Pena C, Johnson C, Canbay A . NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice. Hepatology. 2013; 59(3):898-910. PMC: 4008151. DOI: 10.1002/hep.26592. View

Chi W, Chen H, Li F, Zhu Y, Yin W, Zhuo Y . HMGB1 promotes the activation of NLRP3 and caspase-8 inflammasomes via NF-κB pathway in acute glaucoma. J Neuroinflammation. 2015; 12:137. PMC: 4518626. DOI: 10.1186/s12974-015-0360-2. View

Jin C, Flavell R . Molecular mechanism of NLRP3 inflammasome activation. J Clin Immunol. 2010; 30(5):628-31. DOI: 10.1007/s10875-010-9440-3. View

Liu X, Zhang Z, Ruan J, Pan Y, Magupalli V, Wu H . Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. Nature. 2016; 535(7610):153-8. PMC: 5539988. DOI: 10.1038/nature18629. View

Perz J, Armstrong G, Farrington L, Hutin Y, Bell B . The contributions of hepatitis B virus and hepatitis C virus infections to cirrhosis and primary liver cancer worldwide. J Hepatol. 2006; 45(4):529-38. DOI: 10.1016/j.jhep.2006.05.013. View

Lamkanfi M, Sarkar A, Vande Walle L, Vitari A, Amer A, Wewers M . Inflammasome-dependent release of the alarmin HMGB1 in endotoxemia. J Immunol. 2010; 185(7):4385-92. PMC: 3428148. DOI: 10.4049/jimmunol.1000803. View

Lamkanfi M, Dixit V . Mechanisms and functions of inflammasomes. Cell. 2014; 157(5):1013-22. DOI: 10.1016/j.cell.2014.04.007. View

Watanabe A, Adnan Sohail M, Gomes D, Hashmi A, Nagata J, Sutterwala F . Inflammasome-mediated regulation of hepatic stellate cells. Am J Physiol Gastrointest Liver Physiol. 2009; 296(6):G1248-57. PMC: 2697939. DOI: 10.1152/ajpgi.90223.2008. View

Shao W, Yeretssian G, Doiron K, Hussain S, Saleh M . The caspase-1 digestome identifies the glycolysis pathway as a target during infection and septic shock. J Biol Chem. 2007; 282(50):36321-9. DOI: 10.1074/jbc.M708182200. View

10.

Wang X, Li D, Tao Q, Lin N, Chen Z . A novel hepatitis B virus X-interactive protein: cytochrome C oxidase III. J Gastroenterol Hepatol. 2006; 21(4):711-5. DOI: 10.1111/j.1440-1746.2006.04139.x. View

11.

Paszkowski A, Rau B, Mayer J, Moller P, Beger H . Therapeutic application of caspase 1/interleukin-1beta-converting enzyme inhibitor decreases the death rate in severe acute experimental pancreatitis. Ann Surg. 2001; 235(1):68-76. PMC: 1422397. DOI: 10.1097/00000658-200201000-00009. View

12.

Tang H, Oishi N, Kaneko S, Murakami S . Molecular functions and biological roles of hepatitis B virus x protein. Cancer Sci. 2006; 97(10):977-83. PMC: 11159107. DOI: 10.1111/j.1349-7006.2006.00299.x. View

13.

Duygu F, Karsen H, Aksoy N, Taskin A . Relationship of oxidative stress in hepatitis B infection activity with HBV DNA and fibrosis. Ann Lab Med. 2012; 32(2):113-8. PMC: 3289775. DOI: 10.3343/alm.2012.32.2.113. View

14.

Ivanov A, Valuev-Elliston V, Tyurina D, Ivanova O, Kochetkov S, Bartosch B . Oxidative stress, a trigger of hepatitis C and B virus-induced liver carcinogenesis. Oncotarget. 2016; 8(3):3895-3932. PMC: 5354803. DOI: 10.18632/oncotarget.13904. View

15.

Rahmani Z, Huh K, Lasher R, Siddiqui A . Hepatitis B virus X protein colocalizes to mitochondria with a human voltage-dependent anion channel, HVDAC3, and alters its transmembrane potential. J Virol. 2000; 74(6):2840-6. PMC: 111774. DOI: 10.1128/jvi.74.6.2840-2846.2000. View

16.

Hu L, Chen L, Yang G, Li L, Sun H, Chang Y . HBx sensitizes cells to oxidative stress-induced apoptosis by accelerating the loss of Mcl-1 protein via caspase-3 cascade. Mol Cancer. 2011; 10:43. PMC: 3096594. DOI: 10.1186/1476-4598-10-43. View

17.

Wree A, Marra F . The inflammasome in liver disease. J Hepatol. 2016; 65(5):1055-1056. DOI: 10.1016/j.jhep.2016.07.002. View

18.

Lavanchy D . Hepatitis B virus epidemiology, disease burden, treatment, and current and emerging prevention and control measures. J Viral Hepat. 2004; 11(2):97-107. DOI: 10.1046/j.1365-2893.2003.00487.x. View

19.

Waris G, Huh K, Siddiqui A . Mitochondrially associated hepatitis B virus X protein constitutively activates transcription factors STAT-3 and NF-kappa B via oxidative stress. Mol Cell Biol. 2001; 21(22):7721-30. PMC: 99943. DOI: 10.1128/MCB.21.22.7721-7730.2001. View

20.

Chen X, He W, Hu L, Li J, Fang Y, Wang X . Pyroptosis is driven by non-selective gasdermin-D pore and its morphology is different from MLKL channel-mediated necroptosis. Cell Res. 2016; 26(9):1007-20. PMC: 5034106. DOI: 10.1038/cr.2016.100. View