Table 1.

Studies included in our systematic review that focused on the effects of acute ethanol exposure on cell models.

First Author, Year	Models and Methods	Summary of Effects
Lu et al., 2021 [31]	AML-12 cells treated with 200 mM ethanol over 24 h.	AMPK enhances mitophagy in hepatocytes, and the AMPK–NFE2L2–UQCRC2 axis regulates liver mitophagy.
Chen et al., 2021 [32]	Huh7 cells and murine primary hepatocytes treated with 50–100 mM ethanol over 24 h.	Acute ethanol exposure induces NOX4 and CYP2E1 overexpression and significantly increases autophagy. Antioxidants efficiently block CYP2E1- and NOX4-mediated autophagy induction.
Zhao et al., 2021 [33]	HepG2 cells treated with 0, 100, 200, 300, 400, and 500 mM alcohol over 12 h.	Alcohol induces reversible ferroptosis, which is significantly reduced by ferrostatin-1. Inhibiting autophagy protects HepG2 cells against alcohol-induced ferroptosis by activating the p62−Keap1−Nrf2 pathway.
Liu et al., 2019 [34]	HepG2 cells treated with 200 mmol/L ethanol for 12 h.	Autophagy is inhibited in ethanol-treated HepG2 cells. ALR-expressing HepG2 cells have increased survival rates, improved mitochondrial membrane potential, and increased ATP levels after ethanol treatment. This protection is associated with the upregulation of autophagy markers and downregulation of p62 and mTOR phosphorylation.
You et al., 2018 [35]	Mouse hepatocytes treated with 80 mM ethanol for 6 h.	SNX10 deficiency upregulates LAMP2A expression and CMA activation via Nrf2 and AMPK signaling in vitro, significantly ameliorating ethanol-induced liver damage and hepatic steatosis.
Mahli et al., 2015 [36]	Primary human hepatocytes and HepG2 cells treated with 50 mM alcohol over 16–24 h.	Alcohol and steatosis increase CYP2E1 levels and activity, lipid peroxidation, oxidative stress, pro-inflammatory gene expression, and autophagy via the CYP2E1 and JNK pathways. Autophagy improves the effects of alcohol on lipid accumulation and inflammatory gene expression in liver cells.
Thomes et al., 2013 [37]	Hep G2 cells treated with 50 mM ethanol over 24 h.	Ethanol treatment increases LC3-II expression and decreases its degradation in a dose-dependent manner depending on ADH and CYP2E1 expression. Blocking ethanol oxidation and ROS production prevents the enhancement of LC3-II expression. Direct exposure to acetaldehyde enhances LC3-II content.
Ding et al., 2010 [38]	Murine hepatocytes and HepG2 cells treated with 40, 80, and 160 mM ethanol for 24 h.	Ethanol-induced autophagy requires ethanol metabolism, ROS generation, and mTOR signaling inhibition in vitro. It is selective for cells with damaged mitochondria and accumulated lipid droplets (but not long-lived proteins) and protects cells from ethanol’s toxic effects. Increasing autophagy reduces acute ethanol hepatotoxicity and steatosis.

AMPK, adenosine monophosphate-activated protein kinase; NFE2L2, nuclear factor erythroid 2-related factor 2; UQCRC2, ubiquinol–cytochrome C reductase core protein 2; Huh7, human hepatoma-derived 7 cell line; NOX4, nicotinamide adenine dinucleotide phosphate oxidase 4; HepG2, human epidermoid carcinoma strain 2; CYP2E1, cytochrome P450 2E1; Keap1, Kelch-like associ-ated a protein 1; Nrf2, nuclear factor erythroid 2-related factor 2; ALR, liver regeneration-associated protein; LAMP2A, lysosomal-associated membrane protein 2A; CMA, chaperone-mediated autophagy; JNK, c-Jun N-terminal kinase; LC3-II, microtubule-associated protein light chain 3 II; mTOR, mammalian target of rapamycin complex 1.