{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T18:27:26Z","timestamp":1788805646570,"version":"build-2803163510"},"reference-count":23,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2026,1,9]],"date-time":"2026-01-09T00:00:00Z","timestamp":1767916800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004281","name":"National Science Centre","doi-asserted-by":"publisher","award":["2022\/45\/B\/NZ7\/01056"],"award-info":[{"award-number":["2022\/45\/B\/NZ7\/01056"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Background: Intestinal failure-associated liver disease (IFALD) is a serious complication in patients receiving parenteral nutrition, often exacerbated by inflammation, lipid overload, and oxidative stress. Nobiletin (NOB), a polymethoxylated flavone, is known for its anti-inflammatory and lipid-regulating properties. Methods: We employed an in vitro model using THLE-2 human hepatocytes and primary human cholangiocytes exposed to Intralipid (INT) and lipopolysaccharide (LPS) to simulate IFALD conditions. NOB was tested at non-toxic concentrations (10 and 25 \u00b5M) to assess its protective effects. MTT viability assays, multiplex bead-based immunoassays (MAGPIX), RT-qPCR, and Western blotting were used to evaluate changes in inflammation markers, gene expression, and protein signaling. Moreover, ALT and AST activities were used to assess hepatocellular injury. Results: NOB maintained high cell viability in THLE-2 hepatocytes and cholangiocytes, confirming its low cytotoxicity. NOB normalized ALT and AST activities in both tested cell lines, but the effect reached statistical significance only for ALT in cholangiocytes. Under IFALD-like conditions (LPS+INT), NOB significantly preserved metabolic activity in both cell types. In THLE-2 and cholangiocytes, NOB markedly reduced the phosphorylation of pro-inflammatory proteins JNK, NF-\u03baB, and STAT3, indicating a broad inhibition of inflammatory signaling. Moreover, in THLE-2 cells, NOB upregulated lipid metabolism-related genes (PRKAA2, CYP7A1, and ABCA1) and decreased oxidative stress, thereby enhancing the nuclear translocation of Nrf2 and increasing SOD1 level, which supports the activation of antioxidant defenses. Conclusions: NOB exhibits hepatoprotective properties under IFALD-like conditions in vitro, likely through modulation of inflammation-related signaling and lipid metabolism pathways.<\/jats:p>","DOI":"10.3390\/pharmaceutics18010087","type":"journal-article","created":{"date-parts":[[2026,1,9]],"date-time":"2026-01-09T10:35:52Z","timestamp":1767954952000},"page":"87","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Nobiletin Attenuates Inflammation and Modulates Lipid Metabolism in an In Vitro Model of Intestinal Failure-Associated Liver Disease"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9106-8961","authenticated-orcid":false,"given":"Marta","family":"Belka","sequence":"first","affiliation":[{"name":"Poznan University of Medical Sciences, Doctoral School, Bukowska 70, 60-812 Poznan, Poland"},{"name":"Poznan University of Medical Sciences, Department Pharmaceutical Biochemistry, Rokietnicka 3, 60-806 Poznan, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3401-9503","authenticated-orcid":false,"given":"Aleksandra","family":"Gosty\u0144ska-Stawna","sequence":"additional","affiliation":[{"name":"Poznan University of Medical Sciences, Department of Pharmaceutical Chemistry, Rokietnicka 3, 60-806 Poznan, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8399-7966","authenticated-orcid":false,"given":"Karina","family":"Sommerfeld-Klatta","sequence":"additional","affiliation":[{"name":"Poznan University of Medical Sciences, Department of Toxicology, Rokietnicka 3, 60-806 Poznan, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5106-5937","authenticated-orcid":false,"given":"Maciej","family":"Stawny","sequence":"additional","affiliation":[{"name":"Poznan University of Medical Sciences, Department of Pharmaceutical Chemistry, Rokietnicka 3, 60-806 Poznan, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7275-0298","authenticated-orcid":false,"given":"Violetta","family":"Krajka-Ku\u017aniak","sequence":"additional","affiliation":[{"name":"Poznan University of Medical Sciences, Department Pharmaceutical Biochemistry, Rokietnicka 3, 60-806 Poznan, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.1002\/ncp.11116","article-title":"Intestinal Failure\u2013Associated Liver Disease: Current Challenges in Screening, Diagnosis, and Parenteral Nutrition Considerations","volume":"39","author":"Tabone","year":"2024","journal-title":"Nutr. Clin. Pract."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1007\/s12072-020-10048-8","article-title":"Intestinal Failure-Associated Liver Disease (IFALD): Insights into Pathogenesis and Advances in Management","volume":"14","author":"Lee","year":"2020","journal-title":"Hepatol. Int."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Cheng, Y., Feng, S., Sheng, C., Yang, C., and Li, Y. (2024). Nobiletin from Citrus Peel: A Promising Therapeutic Agent for Liver Disease-Pharmacological Characteristics, Mechanisms, and Potential Applications. Front. Pharmacol., 15.","DOI":"10.3389\/fphar.2024.1354809"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Ke, Z., Fan, C., Li, J., Wang, L., Li, H., Tian, W., and Yu, Q. (2023). Nobiletin Intake Attenuates Hepatic Lipid Profiling and Oxidative Stress in HFD-Induced Nonalcoholic-Fatty-Liver-Disease Mice. Molecules, 28.","DOI":"10.3390\/molecules28062570"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5595","DOI":"10.3892\/mmr.2016.5943","article-title":"Nobiletin Attenuates Lipopolysaccharide\/D-Galactosamine-Induced Liver Injury in Mice by Activating the Nrf2 Antioxidant Pathway and Subsequently Inhibiting NF-\u03baB-Mediated Cytokine Production","volume":"14","author":"He","year":"2016","journal-title":"Mol. Med. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Saha, S., Buttari, B., Panieri, E., Profumo, E., and Saso, L. (2020). An Overview of Nrf2 Signaling Pathway and Its Role in Inflammation. Molecules, 25.","DOI":"10.3390\/molecules25225474"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Mungala Lengo, A., Mohamed, I., and Lavoie, J.-C. (2024). Glutathione Supplementation Prevents Neonatal Parenteral Nutrition-Induced Short- and Long-Term Epigenetic and Transcriptional Disruptions of Hepatic H2O2 Metabolism in Guinea Pigs. Nutrients, 16.","DOI":"10.3390\/nu16060849"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1089\/ars.2021.0076","article-title":"Signal Transduction and Molecular Regulation in Fatty Liver Disease","volume":"35","author":"Dong","year":"2021","journal-title":"Antioxid. Redox Signal."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2138","DOI":"10.1017\/S0007114513001645","article-title":"Betanin, a Beetroot Component, Induces Nuclear Factor Erythroid-2-Related Factor 2-Mediated Expression of Detoxifying\/Antioxidant Enzymes in Human Liver Cell Lines","volume":"110","author":"Paluszczak","year":"2013","journal-title":"Br. J. Nutr."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ab.2014.08.027","article-title":"Stain-Free Detection as Loading Control Alternative to Ponceau and Housekeeping Protein Immunodetection in Western Blotting","volume":"467","author":"Anzola","year":"2014","journal-title":"Anal. Biochem."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Pastor-Clerigues, A., Marti-Bonmati, E., Milara, J., Almudever, P., and Cortijo, J. (2014). Anti-Inflammatory and Anti-Fibrotic Profile of Fish Oil Emulsions Used in Parenteral Nutrition-Associated Liver Disease. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0115404"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1016\/j.jpedsurg.2016.02.078","article-title":"Molecular Impact of Omega 3 Fatty Acids on Lipopolysaccharide-Mediated Liver Damage","volume":"51","author":"Ventro","year":"2016","journal-title":"J. Pediatr. Surg."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Czerniel, J., Gosty\u0144ska-Stawna, A., Sommerfeld-Klatta, K., Przybylski, T., Krajka-Ku\u017aniak, V., and Stawny, M. (2025). Development and Validation of In Vitro Assessment Protocol of Novel Intravenous Nanoemulsions for Parenteral Nutrition. Pharmaceutics, 17.","DOI":"10.3390\/pharmaceutics17040493"},{"key":"ref_14","first-page":"5815","article-title":"Nobiletin Alleviates Palmitic Acid-induced NLRP3 Inflammasome Activation in a Sirtuin 1-dependent Manner in AML-12 Cells","volume":"18","author":"Peng","year":"2018","journal-title":"Mol. Med. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"20105","DOI":"10.1021\/acs.jafc.3c06498","article-title":"Nobiletin Ameliorates Hepatic Lipid Deposition, Oxidative Stress, and Inflammation by Mechanisms That Involve the Nrf2\/NF-\u03baB Axis in Nonalcoholic Fatty Liver Disease","volume":"71","author":"Fan","year":"2023","journal-title":"J. Agric. Food Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"925","DOI":"10.1016\/j.jhep.2014.05.046","article-title":"Strategies for Immortalization of Primary Hepatocytes","volume":"61","author":"Ramboer","year":"2014","journal-title":"J. Hepatol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"E745","DOI":"10.1152\/ajpendo.00015.2018","article-title":"Saturated Fatty Acid Combined with Lipopolysaccharide Stimulates a Strong Inflammatory Response in Hepatocytes in Vivo and in Vitro","volume":"315","author":"Li","year":"2018","journal-title":"Am. J. Physiol.-Endocrinol. Metab."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"9717014","DOI":"10.1155\/2016\/9717014","article-title":"Differential Impacts of Soybean and Fish Oils on Hepatocyte Lipid Droplet Accumulation and Endoplasmic Reticulum Stress in Primary Rabbit Hepatocytes","volume":"2016","author":"Zhu","year":"2016","journal-title":"Gastroenterol. Res. Pract."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1020","DOI":"10.1016\/j.jpedsurg.2017.03.031","article-title":"The Molecular Impact of Omega 3 Fatty Acids on Hepatic Pro-Inflammatory Cytokine Signaling","volume":"52","author":"Ventro","year":"2017","journal-title":"J. Pediatr. Surg."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Machado, I.F., Miranda, R.G., Dorta, D.J., Rolo, A.P., and Palmeira, C.M. (2023). Targeting Oxidative Stress with Polyphenols to Fight Liver Diseases. Antioxidants, 12.","DOI":"10.3390\/antiox12061212"},{"key":"ref_21","first-page":"817","article-title":"The Past and Present of Serum Aminotransferases and the Future of Liver Injury Biomarkers","volume":"15","author":"McGill","year":"2016","journal-title":"EXCLI J."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Gao, W., Guo, L., Yang, Y., Wang, Y., Xia, S., Gong, H., Zhang, B.-K., and Yan, M. (2022). Dissecting the Crosstalk Between Nrf2 and NF-\u03baB Response Pathways in Drug-Induced Toxicity. Front. Cell Dev. Biol., 9.","DOI":"10.3389\/fcell.2021.809952"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"113429","DOI":"10.1016\/j.phytochem.2022.113429","article-title":"Antioxidant and Anti-Inflammation Effects of Dietary Phytochemicals: The Nrf2\/NF-\u03baB Signalling Pathway and Upstream Factors of Nrf2","volume":"204","author":"Wu","year":"2022","journal-title":"Phytochemistry"}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/18\/1\/87\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,9]],"date-time":"2026-01-09T10:39:42Z","timestamp":1767955182000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/18\/1\/87"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,9]]},"references-count":23,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,1]]}},"alternative-id":["pharmaceutics18010087"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics18010087","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,9]]}}}