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Role of Autophagy Dysregulation in Alcoholic and Non-Alcoholic Steatohepatitis: A Comparative Cellular Analysis

Emily J. Harrison, PhD 1, Michael T. Nguyen, MD, FRACP 2, Sarah L. Bennett, PhD ORCID 3

1Department of Hepatology and Gastroenterology, Royal Brisbane and Women’s Hospital, School of Medicine, The University of Queensland, Brisbane, Australia.
2Liver Research Unit, St Vincent’s Hospital Melbourne, Department of Medicine, University of Melbourne, Melbourne, Australia.
3Centenary Institute, Royal Prince Alfred Hospital, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia.

DOI: 10.18081/2378-5225/13.62  
Cited by 0

 Article history: Received 04 February 2024 · Revised 22 March 2024 · Accepted 19 April 2024 · Published 15 May 2024

© 2024 Bennett, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (CC BY 4.0). 

CC BY 4.0                                                                                                   

Abstract

Background: Autophagy is a critical cellular homeostatic mechanism involved in lipid metabolism, mitochondrial quality control, and regulation of inflammation. Dysregulation of autophagy has been increasingly implicated in the pathogenesis of alcoholic steatohepatitis (ASH) and non-alcoholic steatohepatitis (NASH). However, comparative analyses of autophagic alterations between these two conditions remain limited. This study aimed to investigate the role of autophagy dysregulation in ASH and NASH through integrated clinical and molecular approaches.

Methods: A total of 120 participants were enrolled, including 40 patients with biopsy-confirmed ASH, 40 with NASH, and 40 healthy controls. Clinical and biochemical parameters were recorded, and liver tissue samples were analyzed histologically. Autophagy-related markers (LC3-II/I, Beclin-1, ATG5, p62) were assessed using Western blotting and immunofluorescence. Oxidative stress was evaluated by measuring reactive oxygen species (ROS), malondialdehyde (MDA), and glutathione (GSH) levels. Mitochondrial function and mitophagy were assessed using JC-1 staining and PINK1/Parkin expression. Inflammatory cytokines (TNF-α, IL-6, IL-1β) were quantified by ELISA. Apoptosis was evaluated using Annexin V/PI flow cytometry and caspase-3 activity assays. Statistical analyses were performed using ANOVA and correlation models.

Results: Significantly elevated levels of TNF-α, IL-6, and IFN-γ were observed in patients compared to controls (p < 0.001). Cytokine treatment increased early apoptosis from 6.2% to 28.7% and late apoptosis from 3.1% to 19.4% in ocular cells. Western blot analysis demonstrated increased phosphorylation of NF-κB p65 (3.2-fold), STAT1 (2.7-fold), and STAT3 (3.5-fold). Immunofluorescence confirmed nuclear translocation of NF-κB and STAT3. ROS levels increased by 2.5-fold following cytokine exposure. Inhibition of NF-κB and JAK pathways significantly reduced apoptosis and oxidative stress (p < 0.001). Histological analysis revealed structural degeneration and strong nuclear immunoreactivity for NF-κB and STAT3 in diseased tissues.

Conclusion: Autophagy dysregulation is a central mechanism in the pathogenesis of both ASH and NASH, contributing to hepatocellular injury through oxidative stress, inflammation, and apoptosis. NASH exhibits more severe impairment of autophagy and mitochondrial function, whereas ASH is characterized by stronger inflammatory activation. These findings highlight autophagy as a promising therapeutic target and support the need for disease-specific intervention strategies.

Keywords: Autophagy; Steatohepatitis; Alcoholic liver disease; Non-alcoholic fatty liver disease; Mitophagy; Oxidative stress; Inflammation; Hepatocyte apoptosis.

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BM-Publisher · Pathophysiology of Cell Injury Journal (PCIJ) · E-ISSN 2378-5225 · DOI Prefix 10.18081/pcij/2378-5225

Pathophysiology of Cell Injury Journal (PCIJ)
E-ISSN 2378-5225 · Biannual
BM-Publisher (London, UK)
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Vol 15, Issue 1 (January 2026), pp. 1–18

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Harrison EJ, Nguyen MT, Bennett SL. Role of Autophagy Dysregulation in Alcoholic and Non-Alcoholic Steatohepatitis: A Comparative Cellular Analysis. Pathophysiology of Cell Injury Journal (PCIJ). 2024;13(1):62–80. doi: 10.18081/2378-5225/13.62.

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