{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T13:29:52Z","timestamp":1787318992665,"version":"build-2736575974"},"reference-count":49,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2016,6,30]],"date-time":"2016-06-30T00:00:00Z","timestamp":1467244800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Demethyleneberberine (DMB) is an essential metabolite of Berberine (BBR) in vivo. Recent reports have revealed multiple novel therapeutic applications of BBR. However, the pharmacological activities of DMB remain to be elucidated. This study aimed to demonstrate the hepatoprotective and anti-fibrotic effects of DMB both in vitro and in vivo. Here we showed that DMB protects against thioacetamide (TAA)-induced hepatic fibrosis in mice and exhibits a higher safety profile as compared to BBR. Flow cytometry and Western blotting analysis showed that DMB is able to suppress the activation of hepatic stellate cells (HSCs) and induce cell apoptosis through the nuclear factor-\u03baB (NF-\u03baB) cascade. Immunohistochemical (IHC) and quantitative polymerase chain reaction (qPCR) analysis indicated that DMB also has inhibitory effects on collagen synthesis and is able to increase collagen degradation by blocking the transforming growth factor \u03b2 1 (TGF-\u03b21)-Smad signaling and reducing the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of MMP (TIMPs). These findings indicate that DMB has the potential to attenuate hepatic fibrosis via suppressing HSC activation.<\/jats:p>","DOI":"10.3390\/ijms17071036","type":"journal-article","created":{"date-parts":[[2016,6,30]],"date-time":"2016-06-30T14:24:28Z","timestamp":1467296668000},"page":"1036","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":36,"title":["Demethyleneberberine Protects against Hepatic Fibrosis in Mice by Modulating NF-\u03baB Signaling"],"prefix":"10.3390","volume":"17","author":[{"given":"Yongchen","family":"Wang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Natural Medicines, Department of Biochemistry, China Pharmaceutical University, Nanjing 210009, China"},{"name":"Divisions of Molecular Medicine and Cardiology, Departments of Anesthesiology and Medicine, David Geffen School of Medicine at University of California Los Angeles, Los Angeles, CA 90095, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zheng","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Nanjing First Hospital, Nanjing Medical University, Nanjing 210009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Yan","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural Medicines, Department of Biochemistry, China Pharmaceutical University, Nanjing 210009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyan","family":"Qiang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural Medicines, Department of Biochemistry, China Pharmaceutical University, Nanjing 210009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cuisong","family":"Zhou","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural Medicines, Department of Biochemistry, China Pharmaceutical University, Nanjing 210009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruiyan","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural Medicines, Department of Biochemistry, China Pharmaceutical University, Nanjing 210009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huan","family":"Chen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural Medicines, Department of Biochemistry, China Pharmaceutical University, Nanjing 210009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yubin","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural Medicines, Department of Biochemistry, China Pharmaceutical University, Nanjing 210009, China"},{"name":"Department of Biochemistry, China Pharmaceutical University, 24 Tongjia Xiang, Nanjing 210009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2016,6,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1002\/iub.1348","article-title":"Tetramethylpyrazine reduces inflammation in liver fibrosis and inhibits inflammatory cytokine expression in hepatic stellate cells by modulating NLRP3 inflammasome pathway","volume":"67","author":"Wu","year":"2015","journal-title":"IUBMB Life"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1007\/BF01324802","article-title":"The hepatic stellate (Ito) cell: Its role in human liver disease","volume":"430","author":"Hautekeete","year":"1997","journal-title":"Virchows Arch."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"618","DOI":"10.1046\/j.1440-1746.1999.01928.x","article-title":"Liver fibrogenesis and the role of hepatic stellate cells: New insights and prospects for therapy","volume":"14","author":"Li","year":"1999","journal-title":"J. Gastroen. Hepatol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1016\/j.dld.2012.02.015","article-title":"Imidazolium salt attenuates thioacetamide-induced liver fibrosis in mice by modulating inflammation and oxidative stress","volume":"44","author":"Suhaimi","year":"2012","journal-title":"Dig. Liver Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"4223","DOI":"10.1074\/jbc.M110709200","article-title":"Mapping the ligand-binding sites and disease-associated mutations on the most abundant protein in the human, type I collagen","volume":"277","author":"Sweeney","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1085","DOI":"10.1038\/sj.bjp.0705339","article-title":"An angiotensin II type 1 receptor antagonist, olmesartan medoxomil, improves experimental liver fibrosis by suppression of proliferation and collagen synthesis in activated hepatic stellate cells","volume":"139","author":"Kurikawa","year":"2003","journal-title":"Br. J. Pharmacol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"955","DOI":"10.1016\/j.jhep.2007.02.003","article-title":"Expression of MMPs and TIMPs in liver fibrosis\u2014A systematic review with special emphasis on anti-fibrotic strategies","volume":"46","author":"Hemmann","year":"2007","journal-title":"J. Hepatol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1092","DOI":"10.1378\/chest.08-1071","article-title":"Critical management decisions in patients with acute liver failure","volume":"134","author":"Stravitz","year":"2008","journal-title":"Chest"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1038\/labinvest.2008.20","article-title":"Activation of peroxisome proliferator-activated receptor-\u03b3 by curcumin blocks the signaling pathways for PDGF and EGF in hepatic stellate cells","volume":"88","author":"Lin","year":"2008","journal-title":"Lab. Investig."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1016\/j.toxlet.2012.07.019","article-title":"Ameliorative effect of grape seed proanthocyanidin extract on thioacetamide-induced mouse hepatic fibrosis","volume":"213","author":"Li","year":"2012","journal-title":"Toxicol. Lett."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"546","DOI":"10.1006\/bbrc.2000.2993","article-title":"Involvement of p65 in the regulation of NF-\u03baB in rat hepatic stellate cells during cirrhosis","volume":"273","author":"Vasiliou","year":"2000","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"He, X., Pu, G., Tang, R., Zhang, D., and Pan, W. (2014). Activation of nuclear factor \u03baB in the hepatic stellate cells of mice with schistosomiasis japonica. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0104323"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1066","DOI":"10.1056\/NEJM199704103361506","article-title":"Mechanisms of disease\u2014Nuclear factor-\u03baB\u2014A pivotal transcription factor in chronic inflammatory diseases","volume":"336","author":"Barnes","year":"1997","journal-title":"N. Engl. J. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"20727","DOI":"10.1074\/jbc.M111.231381","article-title":"Mapping the I \u03baB kinase \u03b2 (IKK \u03b2)-binding interface of the B14 protein, a vaccinia virus inhibitor of IKK \u03b2-mediated activation of nuclear factor \u03baB","volume":"286","author":"Benfield","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"239","DOI":"10.33549\/physiolres.931437","article-title":"Evaluation of oxidative status in acetaminophen-treated rat hepatocytes in culture","volume":"58","author":"Rousar","year":"2009","journal-title":"Physiol. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1016\/j.lfs.2011.02.009","article-title":"Effect of globin digest on the liver injury and hepatic gene expression profile in galactosamine-induced liver injury in SD rats","volume":"88","author":"Yamamoto","year":"2011","journal-title":"Life Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"6605","DOI":"10.3748\/wjg.v18.i45.6605","article-title":"Hepatoprotective effects of baicalein against CCl4-induced acute liver injury in mice","volume":"18","author":"Huang","year":"2012","journal-title":"World J. Gastroenterol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"626","DOI":"10.1126\/science.108.2814.626","article-title":"Liver tumours in rats fed thiourea or thioacetamide","volume":"108","author":"Fitzhugh","year":"1948","journal-title":"Science"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1877","DOI":"10.1124\/dmd.105.005520","article-title":"Saturation toxicokinetics of thioacetamide: Role in initiation of liver injury","volume":"33","author":"Chilakapati","year":"2005","journal-title":"Drug Metab. Dispos."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1016\/j.taap.2007.11.010","article-title":"Role of CYP2E1 in thioacetamide-induced mouse hepatotoxicity","volume":"228","author":"Kang","year":"2008","journal-title":"Toxicol. Appl. Pharm."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1016\/S0168-8278(96)80167-X","article-title":"Intracellular calcium concentration impairment in hepatocytes from thioacetamide-treated rats. Implications for the activity of Ca2+-dependent enzymes","volume":"24","author":"DiezFernandez","year":"1996","journal-title":"J. Hepatol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2097","DOI":"10.1016\/j.tiv.2010.06.011","article-title":"The toxic effect of thioacetamide on rat liver in vitro","volume":"24","author":"Stankova","year":"2010","journal-title":"Toxicol. In Vitro"},{"key":"ref_23","first-page":"805","article-title":"Histological studies of liver cirrhosis in white rats by thioacetamide feeding","volume":"47","author":"Miyazaki","year":"1956","journal-title":"Gan"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1192","DOI":"10.1038\/labinvest.2008.91","article-title":"Activation of hepatic stellate cells is associated with cytokine expression in thioacetamide-induced hepatic fibrosis in mice","volume":"88","author":"Palacios","year":"2008","journal-title":"Lab. Investig."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"949","DOI":"10.1177\/0192623311418680","article-title":"Role of quercetin in preventing thioacetamide-induced liver injury in rats","volume":"39","author":"Rodrigues","year":"2011","journal-title":"Toxicol. Pathol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1016\/j.ejphar.2011.08.041","article-title":"Comparison of early treatment with low doses of nilotinib, imatinib and a clinically relevant dose of silymarin in thioacetamide-induced liver fibrosis","volume":"670","author":"Shaker","year":"2011","journal-title":"Eur. J. Pharmacol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.mce.2010.07.023","article-title":"Berberine attenuates lipopolysaccharide-induced extracelluar matrix accumulation and inflammation in rat mesangial cells: Involvement of NF-\u03baB signaling pathway","volume":"331","author":"Jiang","year":"2011","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1346","DOI":"10.1158\/1535-7163.MCT-10-0985","article-title":"Berberine suppresses androgen receptor signaling in prostate cancer","volume":"10","author":"Li","year":"2011","journal-title":"Mol. Cancer Ther."},{"key":"ref_29","first-page":"292","article-title":"Berberine inhibits the proliferation of colon cancer cells by inactivating Wnt\/\u03b2-catenin signaling","volume":"41","author":"Wu","year":"2012","journal-title":"Int. J. Oncol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1533","DOI":"10.1248\/bpb.32.1533","article-title":"Berberine inhibits hepatic stellate cell proliferation and prevents experimental liver fibrosis","volume":"32","author":"Sun","year":"2009","journal-title":"Biol. Pharm. Bull."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1124\/jpet.114.219832","article-title":"Demethyleneberberine, a natural mitochondria-targeted antioxidant, inhibits mitochondrial dysfunction, oxidative stress, and steatosis in alcoholic liver disease mouse model","volume":"352","author":"Zhang","year":"2015","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1136\/gut.2005.088690","article-title":"Emerging insights into transforming growth factor \u03b2 Smad signal in hepatic fibrogenesis","volume":"56","author":"Inagaki","year":"2007","journal-title":"Gut"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.lfs.2013.12.211","article-title":"Hepatoprotective effects of berberine on liver fibrosis via activation of AMP-activated protein kinase","volume":"98","author":"Li","year":"2014","journal-title":"Life Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2064","DOI":"10.1124\/dmd.106.011361","article-title":"Pharmacokinetics of berberine and its main metabolites in conventional and pseudo germ-free rats determined by liquid chromatography\/ion trap mass spectrometry","volume":"34","author":"Zuo","year":"2006","journal-title":"Drug Metab. Dispos."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1105","DOI":"10.1016\/j.fct.2010.01.033","article-title":"Acute toxicity of berberine and its correlation with the blood concentration in mice","volume":"48","author":"Kheir","year":"2010","journal-title":"Food Chem. Toxicol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/S0168-8278(99)80160-3","article-title":"The hydroxyl radical scavengers dimethylstioxide and dimethylthiourea protect rats against thioacetamide-induced f-ant hepatic failure","volume":"31","author":"Bruck","year":"1999","journal-title":"J. Heputol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/S0378-4274(00)00168-5","article-title":"Role of metabolic activation by cytochrome P450 in thioacetamide-induced suppression of antibody response in male BALB\/c mice","volume":"114","author":"Kim","year":"2000","journal-title":"Toxicol. Lett."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.lfs.2015.04.010","article-title":"Pituitary tumor transforming gene as a novel regulatory factor of liver fibrosis","volume":"132","author":"Buko","year":"2015","journal-title":"Life Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.taap.2015.09.028","article-title":"Silent information regulator 1 (SIRT1) ameliorates liver fibrosis via promoting activated stellate cell apoptosis and reversion","volume":"289","author":"Wu","year":"2015","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"4095","DOI":"10.3892\/mmr.2015.3992","article-title":"Hepatoprotective effect of juglone on dimethylnitrosamine-induced liver fibrosis and its effect on hepatic antioxidant defence and the expression levels of \u03b1-SMA and collagen III","volume":"12","author":"Zhou","year":"2015","journal-title":"Mol. Med. Rep."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.ejphar.2010.10.025","article-title":"Neferine inhibits cultured hepatic stellate cell activation and facilitates apoptosis A possible molecular mechanism","volume":"650","author":"Ding","year":"2011","journal-title":"Eur. J. Pharmacol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1223","DOI":"10.1111\/jgh.12147","article-title":"Paeonol inhibits hepatic fibrogenesis via disrupting nuclear factor-B pathway in activated stellate cells:in vivo and in vitro studies","volume":"28","author":"Kong","year":"2013","journal-title":"J. Gastroen. Hepatol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.ejps.2014.09.001","article-title":"Curcumol induces HSC-T6 cell death through suppression of Bcl-2: Involvement of PI3K and NF-\u03baB pathways","volume":"65","author":"Chen","year":"2014","journal-title":"Eur. J. Pharm. Sci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/S0168-8278(00)80159-2","article-title":"Nuclear factor \u03baB in proliferation, activation, and apoptosis in rat hepatic stellate cells","volume":"33","author":"Lang","year":"2000","journal-title":"J. Hepatol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"G1059","DOI":"10.1152\/ajpgi.1999.276.4.G1059","article-title":"TGF-\u03b2 1 in liver fibrosis: An inducible transgenic mouse model to study liver fibrogenesis","volume":"276","author":"Kanzler","year":"1999","journal-title":"Am. J. Physiol.-Gastrointest. Liver Physiol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1016\/j.bbrc.2016.03.019","article-title":"Demethyleneberberine attenuates non-alcoholic fatty liver disease with activation of AMPK and inhibition of oxidative stress","volume":"472","author":"Qiang","year":"2016","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1111\/j.1530-0277.1989.tb00276.x","article-title":"Beef fat prevents alcoholic liver disease in the rat","volume":"13","author":"Nanji","year":"1989","journal-title":"Alcohol. Clin. Exp. Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1002\/hep.1840100509","article-title":"The integrated value of serum procollagen III peptide over time predicts hepatic hydroxyproline content and stainable collagen in a model of dietary cirrhosis in the rat","volume":"10","author":"Ruwart","year":"1989","journal-title":"Hepatology"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2212\u0394\u0394Ct method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/17\/7\/1036\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T19:25:15Z","timestamp":1760210715000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/17\/7\/1036"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,6,30]]},"references-count":49,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2016,7]]}},"alternative-id":["ijms17071036"],"URL":"https:\/\/doi.org\/10.3390\/ijms17071036","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,6,30]]}}}