PMID- 32330379 OWN - NLM STAT- MEDLINE DCOM- 20201111 LR - 20211204 IS - 1421-9778 (Electronic) IS - 1015-8987 (Linking) VI - 54 IP - 3 DP - 2020 Apr 25 TI - Genistein and 17beta-Estradiol Protect Hepatocytes from Fatty Degeneration by Mechanisms Involving Mitochondria, Inflammasome and Kinases Activation. PG - 401-416 LID - 10.33594/000000227 [doi] AB - BACKGROUND/AIMS: Oxidative stress and mitochondria dysfunction could be involved in the onset of non-alcoholic fatty liver disease (NAFLD) and in its progression to non-alcoholic steatohepatitis (NASH). Estrogens/phytoestrogens could counteract liver fat deposition with beneficial effects against NAFLD by unclear mechanisms. We aimed to analyze the protective effects elicited by genistein/estradiol in hepatocytes cultured in NAFLD-like medium on cell viability, triglycerides accumulation, mitochondrial function and oxidative stress and the role of NLRP3 inflammasome, toll like receptors 4 (TLR4), Akt and 5' AMP-activated protein kinase (AMPK)alpha1/2. METHODS: Human primary hepatocytes/hepatoma cell line (Huh7.5 cells) were incubated with a 2 mM mixture of oleate/palmitate in presence/absence of genistein/17beta-estradiol. In some experiments, Huh7.5 cells were exposed to various inhibitors of the above pathways and estrogenic receptors (ERs) and G protein-coupled estrogen receptor (GPER) blockers, before genistein/17beta-estradiol. Cell viability, mitochondrial membrane potential, reactive oxygen species and triglycerides content were examined by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT), 5,51,6,61-tetrachloro-1,11,3,31 tetraethylbenzimidazolyl carbocyanine iodide (JC-1), 2,7-dichlorodihydrofluorescein diacetate (H2DCFDA) and the Triglyceride Colorimetric Assay. The expression/activation of kinases was analyzed by means of Western blot. RESULTS: Genistein/17beta-estradiol protected hepatocytes against NAFLD-like medium, by preventing the loss of cell viability and mitochondrial function, triglycerides accumulation and peroxidation. The blocking of kinases, ERs and GPER was able to reduce the above effects, which were potentiated by NLRP3 inflammasome. CONCLUSION: Our findings suggest novel mechanisms underlying the protective effects elicited by phytoestrogens/estrogens against NAFLD/NASH and open novel therapeutic perspectives in the management of NAFLD in postmenopausal women. CI - (c) Copyright by the Author(s). Published by Cell Physiol Biochem Press. FAU - Farruggio, Serena AU - Farruggio S AD - Laboratory of Physiology and Experimental Surgery, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. AD - AGING Project, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. FAU - Cocomazzi, Grazia AU - Cocomazzi G AD - Laboratory of Physiology and Experimental Surgery, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. AD - AGING Project, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. FAU - Marotta, Patrizia AU - Marotta P AD - Laboratory of Physiology and Experimental Surgery, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. AD - AGING Project, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. FAU - Romito, Raffaele AU - Romito R AD - General Surgery Unit, Maggiore della Carita University Hospital, Novara, Italy. FAU - Surico, Daniela AU - Surico D AD - AGING Project, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. AD - Gynecology Division, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. FAU - Calamita, Giuseppe AU - Calamita G AD - Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari "Aldo Moro", Bari, Italy. FAU - Bellan, Mattia AU - Bellan M AD - AGING Project, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. AD - Internal Medicine Division, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. FAU - Pirisi, Mario AU - Pirisi M AD - AGING Project, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. AD - Internal Medicine Division, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. FAU - Grossini, Elena AU - Grossini E AD - Laboratory of Physiology and Experimental Surgery, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy, elena.grossini@med.uniupo.it. AD - AGING Project, Department of Translational Medicine, Universita del Piemonte Orientale, Novara, Italy. LA - eng PT - Journal Article PL - Germany TA - Cell Physiol Biochem JT - Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology JID - 9113221 RN - 0 (GPER1 protein, human) RN - 0 (Inflammasomes) RN - 0 (Phytoestrogens) RN - 0 (Reactive Oxygen Species) RN - 0 (Receptors, Estrogen) RN - 0 (Receptors, G-Protein-Coupled) RN - 0 (TLR4 protein, human) RN - 0 (Toll-Like Receptor 4) RN - 0 (Triglycerides) RN - 4TI98Z838E (Estradiol) RN - DH2M523P0H (Genistein) RN - EC 2.7.- (Protein Kinases) RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) RN - EC 2.7.11.3 (AMP-Activated Protein Kinase Kinases) SB - IM MH - AMP-Activated Protein Kinase Kinases MH - Cell Line MH - Cell Survival/*drug effects MH - Estradiol/*pharmacology MH - Genistein/*pharmacology MH - Hepatocytes/*drug effects/metabolism MH - Humans MH - Inflammasomes/*metabolism MH - Membrane Potential, Mitochondrial/drug effects MH - Mitochondria/*metabolism MH - Non-alcoholic Fatty Liver Disease/*metabolism MH - Oxidative Stress/drug effects MH - Phytoestrogens/pharmacology MH - Protein Kinases/metabolism MH - Proto-Oncogene Proteins c-akt/metabolism MH - Reactive Oxygen Species/metabolism MH - Receptors, Estrogen/antagonists & inhibitors MH - Receptors, G-Protein-Coupled/antagonists & inhibitors MH - Toll-Like Receptor 4/metabolism MH - Triglycerides/metabolism OTO - NOTNLM OT - Cell viability; Mitochondria; NAFLD; Phytoestrogens; Triglycerides COIS- The authors have no conflicts of interest to declare. EDAT- 2020/04/25 06:00 MHDA- 2020/11/12 06:00 CRDT- 2020/04/25 06:00 PHST- 2020/04/11 00:00 [accepted] PHST- 2020/04/25 06:00 [entrez] PHST- 2020/04/25 06:00 [pubmed] PHST- 2020/11/12 06:00 [medline] AID - 10.33594/000000227 [doi] PST - ppublish SO - Cell Physiol Biochem. 2020 Apr 25;54(3):401-416. doi: 10.33594/000000227.