Basic Study
Copyright ©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Hepatol. Jan 27, 2024; 16(1): 75-90
Published online Jan 27, 2024. doi: 10.4254/wjh.v16.i1.75
Rifaximin on epigenetics and autophagy in animal model of hepatocellular carcinoma secondary to metabolic-dysfunction associated steatotic liver disease
Matheus Truccolo Michalczuk, Larisse Longo, Melina Belén Keingeski, Bruno de Souza Basso, Gabriel Tayguara Silveira Guerreiro, Jessica T Ferrari, José Eduardo Vargas, Cláudia P Oliveira, Carolina Uribe-Cruz, Carlos Thadeu Schmidt Cerski, Eduardo Filippi-Chiela, Mário Reis Álvares-da-Silva
Matheus Truccolo Michalczuk, Mário Reis Álvares-da-Silva, Division of Gastroenterology, Hospital de Clínicas de Porto Alegre, Porto Alegre 90035-903, Rio Grande do Sul, Brazil
Matheus Truccolo Michalczuk, Larisse Longo, Melina Belén Keingeski, Gabriel Tayguara Silveira Guerreiro, Jessica T Ferrari, Carolina Uribe-Cruz, Carlos Thadeu Schmidt Cerski, Eduardo Filippi-Chiela, Mário Reis Álvares-da-Silva, Graduate Program in Gastroenterology and Hepatology, Universidade Federal do Rio Grande do Sul, Porto Alegre 90035-003, Rio Grande do Sul, Brazil
Larisse Longo, Melina Belén Keingeski, Bruno de Souza Basso, Gabriel Tayguara Silveira Guerreiro, Carolina Uribe-Cruz, Mário Reis Álvares-da-Silva, Experimental Laboratory of Hepatology and Gastroenterology, Hospital de Clínicas de Porto Alegre, Porto Alegre 90035-903, Rio Grande do Sul, Brazil
José Eduardo Vargas, Laboratory of Inflammatory and Neoplastic Cells, Universidade Federal do Paraná, Paraná 81530900, Brazil
Cláudia P Oliveira, Department of Gastroenterology (LIM07), Faculdade de Medicina da Universidade de São Paulo, São Paulo 01246903, Brazil
Carolina Uribe-Cruz, Facultad de Ciencias de la Salud, Universidad Católica de las Misiones, Posadas, Misiones 3300, Argentina
Carlos Thadeu Schmidt Cerski, Unit of Surgical Pathology, Hospital de Clinicas de Porto Alegre, Porto Alegre 90035-903, Rio Grande do Sul, Brazil
Eduardo Filippi-Chiela, Center of Biotechnology, Universidade Federal do Rio Grande do Sul, Porto Alegre 90035-003, Rio Grande do Sul, Brazil
Eduardo Filippi-Chiela, Department of Morphological Sciences, Universidade Federal do Rio Grande do Sul, Porto Alegre 90.050-170, Rio Grande do Sul, Brazil
Mário Reis Álvares-da-Silva, Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Researcher, Brasília 71.605-001, Brazil
Co-first authors: Matheus Truccolo Michalczuk and Larisse Longo.
Author contributions: Michalczuk MT and Longo L performed the conceptualization, methodology, formal analysis, investigation, data curation, writing of the original draft, writing-review, and editing; Keingeski MB, Guerreiro GTS, Ferrari JT, Filippi-Chiela E, Uribe-Cruz C and Cerski CTS performed the methodology and formal analysis, writing review and editing; Basso BS performed the methodology; Vargas JE performed the analysis; Oliveira CP performed the methodology and writing review; Álvares-da-Silva MR performed the conceptualization, methodology, formal analysis, investigation, data curation, writing of the original draft, writing-review, editing and research fundraising.
Supported by the following Brazilian funding agencies: Financiamento e Incentivo à Pesquisa from Hospital de Clínicas de Porto Alegre (FIPE/HCPA), No. 2021-0105 (to Álvares-da-Silva MR); Coordination for the Improvement of Higher Education Personnel, CAPES/PNPD; and this study was financed in part by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (to Álvares-da-Silva MR).
Institutional review board statement: Institutional review board approval was obtained for this study from the Grupo de Pesquisa em Pós-Graduação – Comissão de Ética em Uso Animal do Hospital de Clinicas de Porto Alegre.
Institutional animal care and use committee statement: All experimental procedures were approved by the Ethics Committee for the Use of Animals protocol No. 2021-0105, in accordance with international guidelines for animal welfare and measures were taken to minimize animal pain and discomfort.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: Technical appendix, statistical code, and dataset available from the corresponding author at marioreis@live.com.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Mário Reis Álvares-da-Silva, MD, PhD, Professor, Division of Gastroenterology, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, n° 2350/ sala 2033, 2° andar Santa Cecília, Porto Alegre 90035-903, Rio Grande do Sul, Brazil. mrsilva@hcpa.edu.br
Received: November 3, 2023
Peer-review started: November 3, 2023
First decision: December 1, 2023
Revised: December 11, 2023
Accepted: January 2, 2024
Article in press: January 2, 2024
Published online: January 27, 2024
Abstract
BACKGROUND

Prevalence of hepatocellular carcinoma (HCC) is increasing, especially in patients with metabolic dysfunction-associated steatotic liver disease (MASLD).

AIM

To investigate rifaximin (RIF) effects on epigenetic/autophagy markers in animals.

METHODS

Adult Sprague-Dawley rats were randomly assigned (n = 8, each) and treated from 5-16 wk: Control [standard diet, water plus gavage with vehicle (Veh)], HCC [high-fat choline deficient diet (HFCD), diethylnitrosamine (DEN) in drinking water and Veh gavage], and RIF [HFCD, DEN and RIF (50 mg/kg/d) gavage]. Gene expression of epigenetic/autophagy markers and circulating miRNAs were obtained.

RESULTS

All HCC and RIF animals developed metabolic-dysfunction associated steatohepatitis fibrosis, and cirrhosis, but three RIF-group did not develop HCC. Comparing animals who developed HCC with those who did not, miR-122, miR-34a, tubulin alpha-1c (Tuba-1c), metalloproteinases-2 (Mmp2), and metalloproteinases-9 (Mmp9) were significantly higher in the HCC-group. The opposite occurred with Becn1, coactivator associated arginine methyltransferase-1 (Carm1), enhancer of zeste homolog-2 (Ezh2), autophagy-related factor LC3A/B (Map1 Lc3b), and p62/sequestosome-1 (p62/SQSTM1)-protein. Comparing with controls, Map1 Lc3b, Becn1 and Ezh2 were lower in HCC and RIF-groups (P < 0.05). Carm1 was lower in HCC compared to RIF (P < 0.05). Hepatic expression of Mmp9 was higher in HCC in relation to the control; the opposite was observed for p62/Sqstm1 (P < 0.05). Expression of p62/SQSTM1 protein was lower in the RIF-group compared to the control (P = 0.024). There was no difference among groups for Tuba-1c, Aldolase-B, alpha-fetoprotein, and Mmp2 (P > 0.05). miR-122 was higher in HCC, and miR-34a in RIF compared to controls (P < 0.05). miR-26b was lower in HCC compared to RIF, and the inverse was observed for miR-224 (P < 0.05). There was no difference among groups regarding miR-33a, miR-143, miR-155, miR-375 and miR-21 (P > 0.05).

CONCLUSION

RIF might have a possible beneficial effect on preventing/delaying liver carcinogenesis through epigenetic modulation in a rat model of MASLD-HCC.

Keywords: Animal model, Autophagy, Epigenetic, Hepatocellular carcinoma, Metabolic dysfunction-associated steatotic liver disease, Rifaximin

Core Tip: Managing metabolic dysfunction-associated steatotic liver disease (MASLD)-hepatocellular carcinoma (HCC) is a clinical challenge, with many unanswered questions, as autophagy and epigenetics appear to contribute to drug resistance. Additionally, the broad-spectrum oral antibiotic Rifaximin influences inflammation, energy metabolism, and fat storage. Utilizing animal models for MASLD-HCC is crucial in understanding pathophysiological mechanisms and potential therapeutic targets. Furthermore, Rifaximin may have a beneficial effect in rats by possibly preventing or delaying hepatic carcinogenesis through epigenetic modulation.