{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,28]],"date-time":"2026-10-28T23:35:44Z","timestamp":1793230544373,"version":"4.5.0"},"reference-count":76,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2022,12,1]],"date-time":"2022-12-01T00:00:00Z","timestamp":1669852800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001711","name":"Swiss National Science Foundation (SNF)","doi-asserted-by":"publisher","award":["40B2-0 187219"],"award-info":[{"award-number":["40B2-0 187219"]}],"id":[{"id":"10.13039\/501100001711","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Swiss Centre for Applied Human Toxicology (SCAHT)","award":["40B2-0 187219"],"award-info":[{"award-number":["40B2-0 187219"]}]},{"name":"Institute for Chemistry and Bioanalytics at the University of Applied Sciences and Arts Northwestern Switzerland (FHNW)","award":["40B2-0 187219"],"award-info":[{"award-number":["40B2-0 187219"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Low-dose methotrexate (MTX) is a standard therapy for rheumatoid arthritis due to its low cost and efficacy. Despite these benefits, MTX has been reported to cause chronic drug-induced liver injury, namely liver fibrosis. The hallmark of liver fibrosis is excessive scarring of liver tissue, triggered by hepatocellular injury and subsequent activation of hepatic stellate cells (HSCs). However, little is known about the precise mechanisms through which MTX causes hepatocellular damage and activates HSCs. Here, we investigated the mechanisms leading to hepatocyte injury in HepaRG and used immortalized stellate cells (hTERT-HSC) to elucidate the mechanisms leading to HSC activation by exposing mono- and co-cultures of HepaRG and hTERT-HSC to MTX. The results showed that at least two mechanisms are involved in MTX-induced toxicity in HepaRG: (i) oxidative stress through depletion of glutathione (GSH) and (ii) impairment of cellular respiration in a GSH-independent manner. Furthermore, we measured increased levels of endoplasmic reticulum (ER) stress in activated HSC following MTX treatment. In conclusion, we established a human-relevant in vitro model to gain mechanistical insights into MTX-induced hepatotoxicity, linked oxidative stress in HepaRG to a GSH-dependent and -independent pathway, and hypothesize that not only oxidative stress in hepatocytes but also ER stress in HSCs contribute to MTX-induced activation of HSCs.<\/jats:p>","DOI":"10.3390\/ijms232315116","type":"journal-article","created":{"date-parts":[[2022,12,2]],"date-time":"2022-12-02T01:44:46Z","timestamp":1669945486000},"page":"15116","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":76,"title":["Methotrexate-Induced Liver Injury Is Associated with Oxidative Stress, Impaired Mitochondrial Respiration, and Endoplasmic Reticulum Stress In Vitro"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7809-6343","authenticated-orcid":false,"given":"Saskia","family":"Schmidt","sequence":"first","affiliation":[{"name":"School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, 4132 Muttenz, Switzerland"},{"name":"Department of Pharmaceutical Sciences, University of Basel, 4056 Basel, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5955-2596","authenticated-orcid":false,"given":"Catherine Jane","family":"Messner","sequence":"additional","affiliation":[{"name":"School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, 4132 Muttenz, Switzerland"},{"name":"Department of Pharmaceutical Sciences, University of Basel, 4056 Basel, Switzerland"},{"name":"Swiss Centre for Applied Human Toxicology (SCAHT), 4055 Basel, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4624-461X","authenticated-orcid":false,"given":"Carine","family":"Gaiser","sequence":"additional","affiliation":[{"name":"School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, 4132 Muttenz, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carina","family":"H\u00e4mmerli","sequence":"additional","affiliation":[{"name":"School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, 4132 Muttenz, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1449-3913","authenticated-orcid":false,"given":"Laura","family":"Suter-Dick","sequence":"additional","affiliation":[{"name":"School of Life Sciences, University of Applied Sciences and Arts Northwestern Switzerland, 4132 Muttenz, Switzerland"},{"name":"Swiss Centre for Applied Human Toxicology (SCAHT), 4055 Basel, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1016\/j.therap.2018.02.004","article-title":"Safety data and withdrawal of hepatotoxic drugs","volume":"76","author":"Babai","year":"2021","journal-title":"Therapies"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1002\/anr.1780320603","article-title":"Methotrexate in rheumatoid arthritis. 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