{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T20:19:49Z","timestamp":1788553189991,"version":"build-2803163510"},"reference-count":42,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,7,2]],"date-time":"2021-07-02T00:00:00Z","timestamp":1625184000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Med."],"abstract":"<jats:p>Changes of lipidic storage, oxidative stress and mitochondrial dysfunction may be involved in the pathogenesis of non-alcoholic fatty liver disease (NAFLD). Although the knowledge of intracellular pathways has vastly expanded in recent years, the role and mechanisms of circulating triggering factor(s) are debated. Thus, we tested the hypothesis that factors circulating in the blood of NAFLD patients may influence processes underlying the disease. Huh7.5 cells\/primary human hepatocytes were exposed to plasma from 12 NAFLD patients and 12 healthy subjects and specific assays were performed to examine viability, H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> and mitochondrial reactive oxygen species (ROS) release, mitochondrial membrane potential and triglycerides content. The involvement of NLRP3 inflammasome and of signaling related to peroxisome-proliferator-activating-ligand-receptor-\u03b3 (PPAR\u03b3), sterol-regulatory-element-binding-protein-1c (SREBP-1c), nuclear-factor-kappa-light-chain-enhancer of activated B cells (NF-kB), and NADPH oxidase 2 (NOX2) was evaluated by repeating the experiments in the presence of NLRP3 inflammasome blocker, MCC950, and through Western blot. The results obtained shown that plasma of NAFLD patients was able to reduce cell viability and mitochondrial membrane potential by about 48 and 24% (<jats:italic>p<\/jats:italic> &amp;lt; 0.05), and to increase H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub>, mitochondrial ROS, and triglycerides content by about 42, 19, and 16% (<jats:italic>p<\/jats:italic> &amp;lt; 0.05), respectively. An increased expression of SREBP-1c, PPAR\u03b3, NF-kB and NOX2 of about 51, 121, 63, and 46%, respectively, was observed (<jats:italic>p<\/jats:italic> &amp;lt; 0.05), as well. Those effects were reduced by the use of MCC950. Thus, in hepatocytes, exposure to plasma from NAFLD patients induces a NAFLD-like phenotype by interference with NLRP3-inflammasome pathways and the activation of intracellular signaling related to SREBP-1c, PPAR\u03b3, NF-kB and NOX2.<\/jats:p>","DOI":"10.3389\/fmed.2021.693997","type":"journal-article","created":{"date-parts":[[2021,7,2]],"date-time":"2021-07-02T05:01:26Z","timestamp":1625202086000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":18,"title":["Exposure to Plasma From Non-alcoholic Fatty Liver Disease Patients Affects Hepatocyte Viability, Generates Mitochondrial Dysfunction, and Modulates Pathways Involved in Fat Accumulation and Inflammation"],"prefix":"10.3389","volume":"8","author":[{"given":"Elena","family":"Grossini","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Divya Praveen","family":"Garhwal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppe","family":"Calamita","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raffaele","family":"Romito","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristina","family":"Rigamonti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosalba","family":"Minisini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlo","family":"Smirne","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela","family":"Surico","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mattia","family":"Bellan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mario","family":"Pirisi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,7,2]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"3313","DOI":"10.1007\/s00018-018-2860-6","article-title":"Molecular mechanisms of hepatic lipid accumulation in non-alcoholic fatty liver disease","volume":"75","author":"Ipsen","year":"2018","journal-title":"Cell Mol Life Sci."},{"key":"B2","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1002\/hep.28431","article-title":"Global epidemiology of non-alcoholic fatty liver disease-meta-analytic assessment of prevalence, incidence, and outcomes","volume":"64","author":"Younossi","year":"2016","journal-title":"Hepatology."},{"key":"B3","doi-asserted-by":"publisher","first-page":"272","DOI":"10.1111\/j.1439-0396.2007.00752.x","article-title":"Liver lipid metabolism","volume":"92","author":"Nguyen","year":"2008","journal-title":"J Anim Physiol Anim Nutr."},{"key":"B4","doi-asserted-by":"publisher","first-page":"952","DOI":"10.1016\/j.jhep.2011.08.025","article-title":"The interaction of hepatic lipid and glucose metabolism in liver diseases","volume":"56","author":"Bechmann","year":"2012","journal-title":"J Hepatol."},{"key":"B5","doi-asserted-by":"publisher","first-page":"9547613","DOI":"10.1155\/2018\/9547613","article-title":"Role of oxidative stress in pathophysiology of non-alcoholic fatty liver disease","volume":"11","author":"Masarone","year":"2018","journal-title":"Oxid Med Cell Longev"},{"key":"B6","doi-asserted-by":"publisher","first-page":"205","DOI":"10.1007\/s10545-008-0841-x","article-title":"Valproic acid metabolism and its effects on mitochondrial fatty acid oxidation: a review","volume":"31","author":"Silva","year":"2008","journal-title":"J Inherit Metab Dis."},{"key":"B7","doi-asserted-by":"publisher","first-page":"610","DOI":"10.1016\/j.clinbiochem.2012.03.024","article-title":"Role of mitochondria in non-alcoholic fatty liver disease-from origin to propagation","volume":"45","author":"Grattagliano","year":"2012","journal-title":"Clin Biochem"},{"key":"B8","doi-asserted-by":"publisher","first-page":"E484","DOI":"10.1152\/ajpendo.00492.2015","article-title":"Lipotoxicity in steatohepatitis occurs despite an increase in tricarboxylic acid cycle activity","volume":"310","author":"Patterson","year":"2016","journal-title":"Am J Physiol Endocrinol Metab."},{"key":"B9","doi-asserted-by":"publisher","first-page":"1","DOI":"10.4254\/wjh.v10.i1.1","article-title":"Role of inflammatory response in liver diseases: therapeutic strategies","volume":"10","author":"Del","year":"2018","journal-title":"World J. Hepatol."},{"key":"B10","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1186\/1471-230X-14-81","article-title":"NOX2-generated oxidative stress is associated with severity of ultrasound liver steatosis in patients with non-alcoholic fatty liver disease","volume":"14","author":"Del","year":"2014","journal-title":"BMC Gastroenterol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"8742","DOI":"10.1038\/s41598-019-45009-1","article-title":"Environmental microcystin targets the microbiome and increases the risk of intestinal inflammatory pathology via NOX2 in underlying murine model of non-alcoholic fatty liver disease","volume":"9","author":"Sarkar","year":"2019","journal-title":"Sci Rep."},{"key":"B12","doi-asserted-by":"publisher","first-page":"1196","DOI":"10.1016\/j.jhep.2006.03.005","article-title":"'Endocrine NAFLD' a hormonocentric perspective of non-alcoholic fatty liver disease pathogenesis","volume":"44","author":"Lonardo","year":"2006","journal-title":"J Hepatol"},{"key":"B13","doi-asserted-by":"publisher","first-page":"115","DOI":"10.1007\/s10517-016-3559-z","article-title":"Mesenchymal-epithelial transition in culture of stromal progenitor cells isolated from the liver of a patient with alcoholic cirrhosis","volume":"162","author":"Kholodenko","year":"2016","journal-title":"Bull Exp Biol Med."},{"key":"B14","doi-asserted-by":"publisher","first-page":"e0138655","DOI":"10.1371\/journal.pone.0138655","article-title":"All-in-one: advanced preparation of human parenchymal and non-parenchymal liver cells","volume":"10","author":"Werner","year":"2015","journal-title":"PloS ONE"},{"key":"B15","doi-asserted-by":"publisher","first-page":"e0191805","DOI":"10.1371\/journal.pone.0191805","article-title":"Simvastatin and metformin inhibit cell growth in hepatitis C virus infected cells via mTOR increasing PTEN and autophagy","volume":"13","author":"Del","year":"2018","journal-title":"PloS ONE."},{"key":"B16","doi-asserted-by":"publisher","first-page":"401","DOI":"10.33594\/000000227","article-title":"Genistein and 17\u03b2-estradiol protect hepatocytes from fatty degeneration by mechanisms involving mitochondria, inflammasome and kinases activation","volume":"54","author":"Farruggio","year":"2020","journal-title":"Cell Physiol Biochem."},{"key":"B17","doi-asserted-by":"publisher","first-page":"1725","DOI":"10.1159\/000479441","article-title":"Anti-vascular endothelial growth factors protect retinal pigment epithelium cells against oxidation by modulating nitric oxide release and autophagy","volume":"42","author":"De","year":"2017","journal-title":"Cell Physiol Biochem."},{"key":"B18","doi-asserted-by":"publisher","first-page":"e0218437","DOI":"10.1371\/journal.pone.0218437","article-title":"Preeclampsia and intrauterine growth restriction: role of human umbilical cord mesenchymal stem cells-trophoblast cross-talk","volume":"14","author":"Surico","year":"2019","journal-title":"PLoS One."},{"key":"B19","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1016\/j.jdermsci.2018.07.007","article-title":"Anti-oxidative effects of 17 \u03b2-estradiol and genistein in human skin fibroblasts and keratinocytes","volume":"92","author":"Savoia","year":"2018","journal-title":"J Dermatol Sci."},{"key":"B20","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1016\/j.yexcr.2018.04.016","article-title":"Hepatitis B virus X protein elevates Parkin-mediated mitophagy through lon peptidase in starvation","volume":"368","author":"Huang","year":"2018","journal-title":"Exp Cell Res."},{"key":"B21","doi-asserted-by":"publisher","first-page":"1051","DOI":"10.1159\/000478752","article-title":"Modulation of oxidative stress by 17 \u03b2-estradiol and genistein in human hepatic cell lines in vitro","volume":"42","author":"Surico","year":"2017","journal-title":"Cell Physiol Biochem"},{"key":"B22","doi-asserted-by":"publisher","first-page":"pii: E978","DOI":"10.3390\/nu10080978","article-title":"Effects of genistein on differentiation and viability of human visceral adipocytes","volume":"10","author":"Grossini","year":"2018","journal-title":"Nutrients."},{"key":"B23","doi-asserted-by":"publisher","first-page":"216","DOI":"10.1016\/j.freeradbiomed.2020.03.009","article-title":"The uricosuric benzbromarone disturbs the mitochondrial redox homeostasis and activates the NRF2 signaling pathway in HepG2 cells","volume":"152","author":"No\u00ebmi","year":"2020","journal-title":"Free Radic Biol Med."},{"key":"B24","doi-asserted-by":"publisher","first-page":"541471","DOI":"10.1155\/2012\/541471","article-title":"Hydrogen peroxide in inflammation: messenger, guide, and assassin","volume":"2012","author":"Wittmann","year":"2012","journal-title":"Adv Hematol."},{"key":"B25","doi-asserted-by":"publisher","first-page":"e0142438","DOI":"10.1371\/journal.pone.0142438","article-title":"NRF2 regulates PINK1 expression under oxidative stress conditions","volume":"10","author":"Murata","year":"2015","journal-title":"PLoS ONE"},{"key":"B26","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1016\/j.cotox.2019.03.001","article-title":"Human in vitro models of non-alcholic fatty liver disease","volume":"16","author":"M\u00fcller","year":"2019","journal-title":"Cur Opin Toxicol"},{"key":"B27","doi-asserted-by":"publisher","first-page":"394","DOI":"10.1177\/1087057111427586","article-title":"High-content imaging technology for the evaluation of drug-induced steatosis using a multiparametric cell-based assay","volume":"17","author":"Donato","year":"2012","journal-title":"J Biomol Screen."},{"key":"B28","doi-asserted-by":"publisher","first-page":"13001","DOI":"10.1128\/JVI.76.24.13001-13014.2002","article-title":"Highly permissive cell lines for subgenomic and genomic hepatitis C virus RNA replication","volume":"76","author":"Blight","year":"2002","journal-title":"J Virol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"886","DOI":"10.1002\/hep.26749","article-title":"Adaptive immune responses triggered by oxidative stress contribute to hepatic inflammation in NASH","volume":"59","author":"Sutti","year":"2014","journal-title":"Hepatology."},{"key":"B30","doi-asserted-by":"publisher","first-page":"1283","DOI":"10.3390\/antiox9121283","article-title":"N-acetyl cysteine targets hepatic lipid accumulation to curb oxidative stress and inflammation in NAFLD: a comprehensive analysis of the literature","volume":"9","author":"Dludla","year":"2020","journal-title":"Antioxidants."},{"key":"B31","doi-asserted-by":"publisher","first-page":"556","DOI":"10.1038\/s41589-019-0277-7","article-title":"MCC950 directly targets the NLRP3 ATP-hydrolysis motif for inflammasome inhibition","volume":"15","author":"Coll","year":"2019","journal-title":"Nature Chem Biol"},{"key":"B32","doi-asserted-by":"publisher","first-page":"2661","DOI":"10.3748\/wjg.v24.i25.2661","article-title":"Apoptosis and non-alcoholic fatty liver diseases","volume":"24","author":"Kanda","year":"2018","journal-title":"World J Gastroenterol"},{"key":"B33","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1186\/s12944-017-0551-1","article-title":"Free fatty acids profile among lean, overweight and obese non-alcoholic fatty liver disease patients: a case\u2014control study","volume":"16","author":"Feng","year":"2017","journal-title":"Lipids Health Dis."},{"key":"B34","doi-asserted-by":"publisher","first-page":"185","DOI":"10.1002\/hep.20283","article-title":"Free fatty acids promote hepatic lipotoxicity by stimulating TNF-alpha expression via a lysosomal pathway","volume":"40","author":"Feldstein","year":"2004","journal-title":"Hepatology."},{"key":"B35","doi-asserted-by":"publisher","first-page":"116","DOI":"10.1016\/j.freeradbiomed.2020.02.025","article-title":"Role of oxidative stress in the pathogenesis of non-alcoholic fatty liver disease","volume":"152","author":"Chen","year":"2020","journal-title":"Free Radic Biol Med."},{"key":"B36","doi-asserted-by":"publisher","first-page":"1501847","DOI":"10.1155\/2018\/1501847","article-title":"Aquaporin membrane channels in oxidative stress, cell signaling, and aging: recent advances and research trends","volume":"2018","author":"Tamma","year":"2018","journal-title":"Oxid Med Cell Longev."},{"key":"B37","doi-asserted-by":"publisher","first-page":"2099","DOI":"10.1016\/j.cellsig.2015.07.011","article-title":"SREBP-1c overactivates ROS-mediated hepatic NF-\u03baB inflammatory pathway in dairy cows with fatty liver","volume":"27","author":"Li","year":"2015","journal-title":"Cell Signal."},{"key":"B38","doi-asserted-by":"publisher","first-page":"909","DOI":"10.1152\/physrev.00026.2013","article-title":"Mitochondrial reactive oxygen species (ROS) and ROS-induced ROS release","volume":"94","author":"Zorov","year":"2014","journal-title":"Physiol Rev."},{"key":"B39","doi-asserted-by":"publisher","first-page":"pii:E2732","DOI":"10.3390\/ijms19092732","article-title":"NLRP3 inflammasome and IL-33: novel players in sterile liver inflammation","volume":"19","author":"Numann","year":"2018","journal-title":"Int J Mol Sci."},{"key":"B40","doi-asserted-by":"publisher","first-page":"6489012","DOI":"10.1155\/2016\/6489012","article-title":"Role of NLRP3 inflammasome in the progression of NAFLD to NASH","volume":"2016","author":"Wan","year":"2016","journal-title":"Can J Gastroenterol Hepatol"},{"key":"B41","doi-asserted-by":"publisher","first-page":"801","DOI":"10.2147\/DMSO.S206558","article-title":"Novel role of NLRP3-inflammasome in regulation of lipogenesis in fasting-induced hepatic steatosis","volume":"12","author":"Liu","year":"2019","journal-title":"Diabetes Metab Syndr Obes."},{"key":"B42","doi-asserted-by":"publisher","first-page":"1367","DOI":"10.1210\/er.2019-00034","article-title":"The liver as an endocrine organ-linking NAFLD and insulin resistance","volume":"40","author":"Watt","year":"2019","journal-title":"Endocr Rev."}],"container-title":["Frontiers in Medicine"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmed.2021.693997\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,7,2]],"date-time":"2021-07-02T05:01:29Z","timestamp":1625202089000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmed.2021.693997\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,2]]},"references-count":42,"alternative-id":["10.3389\/fmed.2021.693997"],"URL":"https:\/\/doi.org\/10.3389\/fmed.2021.693997","relation":{},"ISSN":["2296-858X"],"issn-type":[{"value":"2296-858X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,2]]},"article-number":"693997"}}