Abstract
Alcohol consumption exacerbates the pathogenesis of hepatitis C virus (HCV) infection and aggravates disease consequences in alcohol‐abusing patients. Although the exact reasons by which alcohol consumption affects several cellular pathways in liver cells are not clear, they might be partially attributed to the ability of alcohol to further suppress the innate immunity, modulation of autophagy and also its relationship with reactive oxygen species (ROS) generation. To evaluate these issues, Huh7 cells harboring HCV replicon and Cytochrome p450 (CYP2E1) plasmid were exposed to ethanol and mRNA expression of Beclin-1, interferon-stimulated gene15 (ISG15) genes and HCV NS5B for two different times were relatively quantitated. ROS was determined by flow cytometry. The results showed that alcohol treatment in a short time caused an increase in HCV NS5B and Beclin-1 mRNA and decreased ISG 15 mRNA. Long-lasting alcohol treatment increased ROS production in Huh-7 cells and HCV replication was reduced. In conclusion, acute alcohol treatment might contribute to increase HCV replication by interference in innate immunity and induction of autophagy. Chronic alcohol treatment caused oxidative stress, which disrupts autophagy and thereby increased the rate of Huh7 cell injury.
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References
Albert M, Becares M, Falqui M, Fernandez-Lozano C, Guerra S (2018) ISG15, a small molecule with huge implications: regulation of mitochondrial homeostasis. Viruses. https://doi.org/10.3390/v10110629
Broering R, Zhang X, Kottilil S, Trippler M, Jiang M, Lu M, Gerken G, Schlaak JF (2010) The interferon stimulated gene 15 functions as a proviral factor for the hepatitis C virus and as a regulator of the IFN response. Gut 59:1111–1119. https://doi.org/10.1136/gut.2009.195545
Brown CR, Maclachlan JH, Cowie BC (2017) Addressing the increasing global burden of viral hepatitis. Hepatobiliary Surg Nutr 6:274–276. https://doi.org/10.21037/hbsn.2017.05.02
Bukong TN, Hou W, Kodys K, Szabo G (2013) Ethanol facilitates hepatitis C virus replication via up-regulation of GW182 and heat shock protein 90 in human hepatoma cells. Hepatology 57:70–80. https://doi.org/10.1002/hep.26010
Chen L, Sun J, Meng L, Heathcote J, Edwards AM, Mcgilvray ID (2010) ISG15, a ubiquitin-like interferon-stimulated gene, promotes hepatitis C virus production in vitro: implications for chronic infection and response to treatment. J Gen Virol 91:382–388. https://doi.org/10.1038/cmi.2014.127
Choi J, Lee KJ, Zheng Y, Yamaga AK, Lai MM, Ou JH (2004) Reactive oxygen species suppress hepatitis C virus RNA replication in human hepatoma cells. Hepatology 39:81–89. https://doi.org/10.1002/hep.20001
Chua PK, Mccown MF, Rajyaguru S, Kular S, Varma R, Symons J, Chiu SS, Cammack N, Najera I (2009) Modulation of alpha interferon anti-hepatitis C virus activity by ISG15. J Gen Virol 90:2929–2939. https://doi.org/10.1099/vir.0.013128-0
Ciccarone F, Castelli S, Ciriolo MR (2019) Oxidative stress-driven autophagy acROSs onset and therapeutic outcome in hepatocellular carcinoma. Oxid Med Cell Longev 2019:6050123. https://doi.org/10.1155/2019/6050123
Ding WX, Li M, Chen X, Ni HM, Lin CW, Gao W, Lu B, Stolz DB, Clemens DL, Yin XM (2010) Autophagy reduces acute ethanol-induced hepatotoxicity and steatosis in mice. Gastroenterology 139:1740–1752. https://doi.org/10.1053/j.gastro.2010.07.041
Dolganiuc A, Thomes PG, Ding WX, Lemasters JJ, Donohue TM Jr (2012) Autophagy in alcohol-induced liver diseases. Alcohol Clin Exp Res 36:1301–1308. https://doi.org/10.1111/j.1530-0277.2012.01742.x
Domingues P, Bamford CG, Boutell C, Mclauchlan J (2015) Inhibition of hepatitis C virus RNA replication by ISG15 does not require its conjugation to protein substrates by the HERC5 E3 ligase. J Gen Virol 96:3236. https://doi.org/10.1099/jgv.0.000283
Dreux M, Chisari FV (2010) Viruses and the autophagy machinery. Cell Cycle 9:1295–1307. https://doi.org/10.4161/cc.9.7.11109
Dreux M, Gastaminza P, Wieland SF, Chisari FV (2009) The autophagy machinery is required to initiate hepatitis C virus replication. Proc Natl Acad Sci 106:14046–14051. https://doi.org/10.4161/auto.5.8.10219
Evans MJ, Rice CM, Goff SP (2004) Genetic interactions between hepatitis C virus replicons. J Virol 78:12085–12089. https://doi.org/10.1128/JVI.78.21.12085-12089.2004
Ganesan M, Zhang J, Bronich T, Poluektova LI, Donohue TM Jr, Tuma DJ, Kharbanda KK, Osna NA (2015) Acetaldehyde accelerates HCV-induced impairment of innate immunity by suppressing methylation reactions in liver cells. Am J Physiol Gastrointest Liver Physiol 309:G566–G577. https://doi.org/10.1111/acer.13226
Ganesan M, Poluektova LY, Tuma DJ, Kharbanda KK, Osna NA (2016) Acetaldehyde disrupts interferon alpha signaling in hepatitis C virus-infected liver cells by up-regulating USP 18. Alcohol Clin Exp Res 40:2329–2338. https://doi.org/10.1152/ajpgi.00183.2015
Geddawy A, Ibrahim YF, Elbahie NM, Ibrahim MA (2017) Direct acting anti-hepatitis C virus drugs: clinical pharmacology and future direction. J Transl Int Med 5:8–17. https://doi.org/10.1515/jtim-2017-0007
Ivanov AV, Smirnova OA, Petrushanko IY, Ivanova ON, Karpenko IL, Alekseeva E, Sominskaya I, Makarov AA, Bartosch B, Kochetkov SN, Isaguliants MG (2015) HCV core protein uses multiple mechanisms to induce oxidative stress in human hepatoma Huh7 cells. Viruses 7:2745–2770. https://doi.org/10.18632/oncotarget.13904
Ivanov AV, Valuev-Elliston VT, Tyurina DA, Ivanova ON, Kochetkov SN, Bartosch B, Isaguliants MG (2017) Oxidative stress, a trigger of hepatitis C and B virus-induced liver carcinogenesis. Oncotarget 8:3895. https://doi.org/10.18632/oncotarget.13904
Kato T, Date T, Murayama A, Morikawa K, Akazawa D (2006) Cell culture and infection system for hepatitis C virus. Nat Protoc 1:2334. https://doi.org/10.1038/nprot.2006.395
Kharbanda KK, Mcvicker DL, Zetterman RK, Donohue TM Jr (1996) Ethanol consumption alters trafficking of lysosomal enzymes and affects the processing of procathepsin L in rat liver. Biochim Biophys Acta Gen Sub 1291:45–52. https://doi.org/10.1016/0304-4165(96)00043-8
Kim MJ, Yoo JY (2010) Inhibition of hepatitis C virus replication by IFN-mediated ISGylation of HCV-NS5A. J Immunol 185:4311–4318. https://doi.org/10.4049/jimmunol.1000098
Lin C-W, Zhang H, Li M, Xiong X, Chen X, Chen X, Dong XC (2013) Pharmacological promotion of autophagy alleviates steatosis and injury in alcoholic and non-alcoholic fatty liver conditions in mice. J Hepatol 58:993–999. https://doi.org/10.1016/j.jhep.2013.01.011
Marascio N, Torti C, Liberto M (2014) Update on different aspects of HCV variability: focus on NS5B polymerase. BMC Infect Dis 14(Suppl 5):S1. https://doi.org/10.1186/1471-2334-14-S5-S1
Matsushita H, Takaki A (2019) Alcohol and hepatocellular carcinoma. BMJ Open Gastroenterol 6:e000260. https://doi.org/10.1136/bmjgast-2018-000260
Mccartney EM, Semendri L, Helbig KJ, Hinze S, Jones B, Weinman SA, Beard MR (2008) Alcohol metabolism increases the replication of hepatitis C virus and attenuates the antiviral action of interferon. J Infect Dis 198:1766–1775. https://doi.org/10.1086/593216
McCartney EM, Beard MR (2010) Impact of alcohol on hepatitis C virus replication and interferon signaling. World J Gastroenterol WJG 16:1337. https://doi.org/10.3748/wjg.v16.i11.1337
Menk M, Graw JA, Poyraz D, Mobius N, Spies CD (2018) Chronic alcohol consumption inhibits autophagy and promotes apoptosis in the liver. Int J Med Sci 15:682–688. https://doi.org/10.7150/ijms.25393
Moriya K, Nakagawa K, Santa T, Shintani Y, Fujie H, Miyoshi H, Tsutsumi T, Miyazawa T, Ishibashi K (2001) Oxidative stress in the absence of inflammation in a mouse model for hepatitis C virus-associated hepatocarcinogenesis. Can Res 61:4365–4370
Muga R, Sanvisens A, Jarrin I, Fuster D, Bolao F, Tor J (2018) Hepatitis C infection substantially reduces survival of alcohol-dependent patients. Clin Epidemiol 10:897–905. https://doi.org/10.2147/clep.S162308
Ohashi K, Pimienta M (2018) Alcoholic liver disease: A current molecular and clinical perspective. Liver Res 2:161–172. https://doi.org/10.1016/j.livres.2018.11.002
Osna N, Ganesan M (2015) Hepatitis C, innate immunity and alcohol: friends or foes? Biomolecules 5:76–94. https://doi.org/10.3390/biom5010076
Osna NA, Thomes PG (2011) Involvement of autophagy in alcoholic liver injury and hepatitis C pathogenesis. World J Gastroenterol WJG 17:2507. https://doi.org/10.3748/wjg.v17.i20.2507
Patra T, Ray RB (2019) Strategies to circumvent host innate immune response by hepatitis C virus. Cells. https://doi.org/10.3390/cells8030274
Plumlee CR, Lazaro CA, Fausto N, Polyak SJ (2005) Effect of ethanol on innate antiviral pathways and HCV replication in human liver cells. Virol J 2:89. https://doi.org/10.1186/1743-422X-2-89
Ran M, Chen H, Liang B, Liao W, Jiang J, Huang J, Ning C, Zang N, Zhou B (2018) Alcohol-induced autophagy via upregulation of PIASy promotes HCV replication in human hepatoma cells. Cell Death Dis 9:898. https://doi.org/10.1038/s41419-018-0845-x
Robinson M, Yang H, Sun SC, Peng B, Tian Y, Pagratis N, Greenstein AE, Delaney WET (2010) Novel hepatitis C virus reporter replicon cell lines enable efficient antiviral screening against genotype 1a. Antimicrob Agents Chemother 54:3099–3106. https://doi.org/10.1128/aac.00289-10
Sir D, Kuo C-F, Tian Y, Liu HM, Huang EJ, Jung JU, Machida K, Ou J-HJ (2012) Replication of hepatitis C virus RNA on autophagosomal membranes. J Biol Chem 287:18036–18043. https://doi.org/10.1074/jbc.M111.320085
Siu L, Foont J, Wands JR (2009) Hepatitis C virus and alcohol. Seminars in liver disease. Thieme Medical Publishers 188–199. Doi 10.1055/s-0029-1214374
Smirnova OA, Ivanova ON, Bartosch B, Valuev-Elliston VT, Mukhtarov F, Kochetkov SN, Ivanov AV (2016) Hepatitis C virus NS5A protein triggers oxidative stress by inducing NADPH oxidases 1 and 4 and cytochrome P450 2E1. Oxid Med Cell Longev 2016:8341937. https://doi.org/10.1155/2016/8341937
Szabo G, Aloman C, Polyak SJ, Weinman SA, Wands J, Zakhari S (2006) Hepatitis C infection and alcohol use: a dangerous mix for the liver and antiviral immunity. Alcohol Clin Exp Res 30:709–719. https://doi.org/10.1111/j.1530-0277.2006.00083.x
Szabo G, Wands JR, Eken A, Osna NA, Weinman SA, Machida K (2010) Alcohol and hepatitis C virus–interactions in immune dysfunctions and liver damage. Alcohol Clin Exp Res 34:1675–1686. https://doi.org/10.1111/j.1530-0277.2006.00083.x
Thomes PG, Trambly CS, Fox HS, Tuma DJ (2015) Acute and chronic ethanol administration differentially modulate hepatic autophagy and transcription factor EB. Alcohol Clin Exp Res 39:2354–2363. https://doi.org/10.1111/acer.12904
Tsutsumi T, Suzuki T, Moriya K, Shintani Y, Fujie H, Miyoshi H, Matsuura Y, Koike K (2003) Hepatitis C virus core protein activates ERK and p38 MAPK in cooperation with ethanol in transgenic mice. Hepatology 38:820–828. https://doi.org/10.1053/jhep.2003.50399
Wang L, Tian Y, Ou J-HJ (2015) HCV induces the expression of Rubicon and UVRAG to temporally regulate the maturation of autophagosomes and viral replication. PLoS Pathog 11:e1004764. https://doi.org/10.1371/journal.ppat.1004764
WHO (2017) Global hepatitis report 2017. World Health Organization, Geneva
Wu D, Wang X, Zhou R (2010) CYP2E1 enhances ethanol-induced lipid accumulation but impairs autophaghy in HepG2 E47 cells. Biochem Biophys Res Commun 402:116–122. https://doi.org/10.1016/j.bbrc.2010.09.127
Zhang T, Li Y, Lai J-P, Douglas SD, Metzger DS, O'Brien CP (2003) Alcohol potentiates hepatitis C virus replicon expression. Hepatology 38:57–65. https://doi.org/10.1053/jhep.2003.50295
Acknowledgements
We gratefully acknowledge Prof. Charles M. Rice for generously providing sub-genomic HCV replicon. We thank Dr. R. Shokohi at the Asthma and Allergy Institute of Imam Khomeini Hospital for statistical analysis.
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This study was funded by the Tarbiat Modares University (Grant Number Med—54395) for Ph.D. thesis.
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FS: project development, data collection, manuscript writing, data analysis. TB: supervising, manuscript editing. FR: advisor.
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Communicated by Erko Stackebrandt.
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Sobhanimonfared, F., Bamdad, T. & Roohvand, F. Cross talk between alcohol-induced oxidative stress and HCV replication. Arch Microbiol 202, 1889–1898 (2020). https://doi.org/10.1007/s00203-020-01909-9
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DOI: https://doi.org/10.1007/s00203-020-01909-9