Table 4.

Studies included in our systematic review that focused on the effects of chronic ethanol intake and exposure on animal models.

First Author, Year	Models and Methods	Summary of Effects
Lu et al., 2021 [31]	NIAAA model with male C57BL/6J mice.	Alcohol induces low UQCRC2 expression, which is alleviated by AMPK. The AMPK–NFE2L2–UQCRC2 axis regulates liver mitophagy.
Guo et al., 2021 [50]	Lieber–DeCarli model with male C57BL/6J mice. Liver tissues from patients with alcoholic hepatitis were examined.	Palmitic acid in alcohol-fed mice induces ER stress and mTORC1-dependent LAMP2 suppression. mTORC1 signaling induction and CHOP were detected in patient livers.
Guo et al., 2021 [51]	Lieber–DeCarli model with male C57BL/6J mice. Liver tissues from patients with alcoholic hepatitis were examined.	The pathogenesis of ALD is mediated by hepatic free fatty acid accumulation, which suppresses the LAMP2–autophagy flux pathway through ER stress signaling.
Babuta et al., 2019 [52]	Lieber–DeCarli and NIAAA models with female C57BL/6 mice. Liver tissues from patients with alcoholic hepatitis were examined.	Chronic alcohol intake impairs autophagy in livers, decreasing mTOR and Rheb and increasing Beclin-l and Atg7 expression, disrupting autophagy at the lysosomal level by decreasing LAMP1 and LAMP2 expression. Alcohol increases miR-155 targeting of mTOR, Rheb, LAMP1, and LAMP2. miR-155-deficient mice are protected from alcohol-induced autophagy disruption and have attenuated exosome production. LAMP1/2 downregulation increases exosome release in hepatocytes in the presence and absence of alcohol.
Zhou et al., 2019 [53]	Female C57BL mice fed liquid diet with 4% (vol/vol) alcohol for 16 weeks.	Chronic alcohol consumption increases DNA-PKcs in the liver, leading to liver damage and mitochondrial dysfunction through p53 activation and defective mitophagy.
Yan et al., 2019 [43]	NIAAA model with male/female C57BL/6 mice.	Mice lacking the Atg7 gene had more liver damage from alcohol and were more susceptible to chronic plus binge drinking. Long-term autophagy deficiency worsened the liver’s response to alcohol.
Menk et al., 2018 [54]	Lieber–DeCarli model with male Wistar rats for 12 weeks.	Chronic alcohol consumption causes stress in the liver, impairs autophagy-related gene expression, disrupts autophagic flux, and increases apoptosis in the liver.
You et al., 2018 [35]	Lieber–DeCarli model with male FVB and C57BL/6J mice for 4 weeks.	SNX10 deficiency increases LAMP2A expression and CMA activation, improving liver damage and fat accumulation caused by alcohol through the activation of Nrf2 and AMPK signaling.
Chao et al., 2018 [55]	NIAAA model with male C57BL/6N mice. Liver tissues from patients with alcoholic hepatitis were examined.	Alcohol-fed mice had lower levels of TFEB, decreased lysosome and autophagy activity, and increased mTOR activation. Activating the TFEB pathway reversed these effects. Mice lacking TFEB or both TFEB and TFE3 had more severe liver damage from alcohol. Patient liver tissues had lower levels of nuclear TFEB than control tissues.
Kong et al., 2017 [56]	Lieber–DeCarli model with male C57BL6 mice for 16 days with intraperitoneal LPS injection (10 mg/kg) on the final day.	Alcohol-fed mice experienced fat accumulation, liver damage, and increased inflammation. LPS worsened alcohol-induced oxidative stress and reduced autophagy activity.
Williams et al., 2015 [44]	NIAAA model with male C57BL/6J mice.	Parkin prevented liver damage in chronic alcohol-fed mice. Mice lacking Parkin had severe mitochondrial damage, reduced mitophagy and mitochondrial function, and an impaired ability to adapt to alcohol treatment.
Guo et al., 2015 [39]	Female FVB mice fed a 4% (vol/vol) alcohol liquid diet for 6 weeks.	Chronic alcohol intake causes liver damage, disturbed fat metabolism, increased inflammation and oxidative stress, and decreased autophagy. Expressing the ALDH2 gene reduced these effects. Lysosomal inhibitors had the same effects as alcohol on p62 accumulation.
Lu and Cederbaum, 2015 [57]	Lieber–DeCarli model with male SV/129 mice for 4 weeks.	Inhibiting autophagy increased liver damage and fat accumulation in mice with normal or increased CYP2E1 levels but not in mice lacking CYP2E1. Autophagy did not affect CYP2E1 activity or induction by alcohol. Mice with normal or increased CYP2E1 levels had decreased autophagy-related gene expression and increased p62 levels.
King et al., 2014 [58]	Lieber–DeCarli model with male C57BL/6J mice.	Alcohol-treated mice experienced fat accumulation, increased autophagy, decreased mitochondrial function, and increased CypD levels. Their mitochondria were more sensitive to damage than those of mice lacking CypD. CypD deficiency impaired autophagy but did not prevent fat accumulation caused by alcohol.
Tan et al., 2013 [59]	Male C57/BL6 mice fed 5–20% (vol/vol) ethanol and a high-fat diet for 8 weeks.	Mice lacking the HFE gene had liver damage, fibrosis, and increased cell death. Iron overload in these mice caused stress responses and impaired autophagy-related gene expression and activity.
Lin et al., 2013 [47]	Lieber–DeCarli model with C57BL/6 mice for 4 weeks.	Macroautophagy was activated during chronic alcohol consumption. Inhibiting autophagy worsened liver damage and fat accumulation, while activating autophagy improved these conditions.
Wu et al., 2012 [40]	Male SV129 mice gavaged with a total of 3 g/kg body weight ethanol over 4 days.	Alcohol treatment caused liver damage, increased CYP2E1 levels, and oxidative stress in mice with normal or increased CYP2E1 levels but not in mice lacking CYP2E1. Alcohol impaired autophagy in mice with increased CYP2E1 levels. Inhibiting autophagy worsened alcohol-induced liver damage, fat accumulation, and oxidative stress in these mice.
Thomes et al., 2012 [49]	Lieber–DeCarli model with GFP-LC3 tg mice for 4–6 weeks.	Chronic alcohol-fed mice had reduced proteasome activity and increased autophagy markers in liver cells. Inhibiting the proteasome further increased autophagy markers.

UQCRC2, ubiquinol–cytochrome C reductase core protein 2; AMPK, adenosine monophosphate-activated protein kinase; NFE2L2, nuclear factor erythroid 2-related factor 2; mTORC1, mammalian target of rapamycin complex 1; LAMP1/2/2A, lysosomal-associated membrane protein 1/2/2A; CHOP, C/EBP Homologous Protein; Rheb, Ras homolog enriched in brain; Atg7, antithymocyte globulin 7; miR, micro-RNA; DNA-PKCs, DNA-dependent protein kinase catalytic subunit; SNX10, sorting nexin-10; CMA, chaperone-mediated autophagy; NRF2, nuclear factor erythroid 2-related factor 2; TFEB, transcription factor EB; TFE3, transcription factor binding to IGHM enhancer 3; LPS, lipopolysaccharide; ALDH2, aldehyde dehydrogenase 2; CYP2E1, cytochrome P450 2E1; CypD, cyclophilin D; HFE, hemochromatosis; GFP, green fluorescent protein; LC3, microtubule-associated protein light chain 3.