{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,17]],"date-time":"2026-06-17T16:06:15Z","timestamp":1781712375132,"version":"3.54.5"},"reference-count":37,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2022,6,1]],"date-time":"2022-06-01T00:00:00Z","timestamp":1654041600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Science and Technology of Taiwan","award":["108-2811-B-002-542"],"award-info":[{"award-number":["108-2811-B-002-542"]}]},{"name":"Ministry of Science and Technology of Taiwan","award":["108-2320-B-002-032-MY"],"award-info":[{"award-number":["108-2320-B-002-032-MY"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Bromelain, an enzyme extracted from the stems of pineapples, exerts anticoagulant effects; however, the regulatory mechanisms are not fully understood. Here, we aimed to investigate the effects of bromelain on non-alcoholic fatty liver disease (NAFLD)-induced deregulation of blood coagulation and the underlying molecular mechanisms. C57BL\/6 mice were fed a high-fat diet (HFD), with or without bromelain (20 mg\/kg\/day) administration, for 12 weeks. Treatment with bromelain decreased thrombus formation in the liver and prolonged HFD-induced shortened prothrombin, activated partial thromboplastin, and fibrinogen times. Moreover, liquid chromatography-mass spectrometry\/mass spectrometry and Western blot analysis showed that bromelain inhibited NAFLD-induced activation of the intrinsic, extrinsic, and common pathways by upregulating the protein expression of antithrombin III, serpin family G member 1, and \u03b11-antitrypsin, and downregulating the protein expression of fibrinogen in the liver and plasma. Bromelain also upregulated the level of plasminogen and downregulating factor XIII expression in the liver and plasma. Collectively, these findings suggest that bromelain exerts anticoagulant effects on NAFLD-induced deregulation of coagulation by inhibiting the activation of the coagulation cascade, decreasing the stability of clots, and promoting fibrinolytic activity. The present study provides new insights into the potential therapeutic value of bromelain for the prevention and treatment of thrombosis-related diseases.<\/jats:p>","DOI":"10.3390\/nu14112329","type":"journal-article","created":{"date-parts":[[2022,6,1]],"date-time":"2022-06-01T12:48:31Z","timestamp":1654087711000},"page":"2329","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["New Mechanisms of Bromelain in Alleviating Non-Alcoholic Fatty Liver Disease-Induced Deregulation of Blood Coagulation"],"prefix":"10.3390","volume":"14","author":[{"given":"Po-An","family":"Hu","sequence":"first","affiliation":[{"name":"Department and Graduate Institute of Physiology, College of Medicine, National Taiwan University, Taipei 10051, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sz-Han","family":"Wang","sequence":"additional","affiliation":[{"name":"Department and Graduate Institute of Physiology, College of Medicine, National Taiwan University, Taipei 10051, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chia-Hui","family":"Chen","sequence":"additional","affiliation":[{"name":"Department and Graduate Institute of Physiology, College of Medicine, National Taiwan University, Taipei 10051, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bei-Chia","family":"Guo","sequence":"additional","affiliation":[{"name":"Department and Graduate Institute of Physiology, College of Medicine, National Taiwan University, Taipei 10051, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jenq-Wen","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, National Taiwan University Hospital, Taipei 10051, Taiwan"},{"name":"Department of Internal Medicine, College of Medicine, National Taiwan University, Taipei 10051, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9593-4062","authenticated-orcid":false,"given":"Tzong-Shyuan","family":"Lee","sequence":"additional","affiliation":[{"name":"Department and Graduate Institute of Physiology, College of Medicine, National Taiwan University, Taipei 10051, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1016\/j.ijbiomac.2016.05.107","article-title":"Amyloidogenic behavior of different intermediate state of stem bromelain: A biophysical insight","volume":"91","author":"Zaman","year":"2016","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1007\/s12325-016-0444-0","article-title":"A randomized, clinical trial to evaluate efficacy and tolerability of trypsin: Chymotrypsin as compared to serratiopeptidase and trypsin: Bromelain: Rutoside in wound management","volume":"34","author":"Chandanwale","year":"2017","journal-title":"Adv. Ther."},{"key":"ref_3","first-page":"121","article-title":"Therapeutic uses of pineapple-extracted bromelain in surgical care-A review","volume":"67","author":"Muhammad","year":"2017","journal-title":"J. Pak. Med. Assoc."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1386","DOI":"10.1002\/jsfa.8122","article-title":"Bromelain: From production to commercialisation","volume":"97","author":"Ramli","year":"2017","journal-title":"J. Sci. Food Agric."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1002\/btpr.2190","article-title":"Stability, purification, and applications of bromelain: A review","volume":"32","author":"Jozala","year":"2016","journal-title":"Biotechnol. Prog."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1097\/MBC.0000000000000531","article-title":"Effects on fibrinogen, fibrin, and blood coagulation of proteolytic extracts from fruits of Pseudananas macrodontes, Bromelia balansae, and B. hieronymi (Bromeliaceae) in comparison with bromelain","volume":"27","author":"Errasti","year":"2016","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"10","DOI":"10.3181\/00379727-93-22647","article-title":"Fibrinolytic and coagulant activities of certain snake venoms and proteases","volume":"93","author":"Didisheim","year":"1956","journal-title":"Proc. Soc. Exp. Biol Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/0378-8741(88)90127-4","article-title":"Bromelain, the enzyme complex of pineapple (Ananas comosus) and its clinical application. An update","volume":"22","author":"Taussig","year":"1988","journal-title":"J. Ethnopharmacol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Moreno-Fern\u00e1ndez, S., Garc\u00e9s-Rim\u00f3n, M., Vera, G., Astier, J., Landrier, J.F., and Miguel, M. (2018). High Fat\/High Glucose Diet Induces Metabolic Syndrome in an Experimental Rat Model. Nutrients, 10.","DOI":"10.3390\/nu10101502"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1242\/dmm.001180","article-title":"A comprehensive definition for metabolic syndrome","volume":"2","author":"Huang","year":"2009","journal-title":"Dis. Model. Mech."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"978","DOI":"10.1016\/j.amjmed.2005.03.012","article-title":"Obesity as a risk factor in venous thromboembolism","volume":"118","author":"Stein","year":"2005","journal-title":"Am. J. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1111\/jth.13892","article-title":"Joint effects of prothrombotic genotypes and body height on the risk of venous thromboembolism: The Troms\u00f8 study","volume":"16","author":"Horvei","year":"2018","journal-title":"J. Thromb. Haemost."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1002\/hep.26510","article-title":"Prothrombotic factors in histologically proven nonalcoholic fatty liver disease and nonalcoholic steatohepatitis","volume":"59","author":"Verrijken","year":"2014","journal-title":"Hepatology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/j.jhep.2016.09.025","article-title":"Procoagulant imbalance in patients with non-alcoholic fatty liver disease","volume":"66","author":"Tripodi","year":"2017","journal-title":"J. Hepatol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1111\/j.1478-3231.2010.02375.x","article-title":"Increased coagulation factor VIII, IX, XI and XII activities in non-alcoholic fatty liver disease","volume":"31","author":"Kotronen","year":"2011","journal-title":"Liver Int."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1111","DOI":"10.1111\/liv.14417","article-title":"Association between plasminogen activator inhibitor-1 in young adulthood and nonalcoholic fatty liver disease in midlife: Cardia","volume":"40","author":"Campbell","year":"2020","journal-title":"Liver Int."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"601","DOI":"10.1016\/j.cvsm.2016.11.009","article-title":"Hemostatic disorders associated with hepatobiliary disease","volume":"47","author":"Webster","year":"2017","journal-title":"Vet. Clin. N. Am. Small Anim. Pract."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.thromres.2020.12.002","article-title":"Translational insight into prothrombotic state and hypercoagulation in nonalcoholic fatty liver disease","volume":"198","author":"Ciavarella","year":"2021","journal-title":"Thromb. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1093\/labmed\/lmx050","article-title":"Coagulation testing in the core laboratory","volume":"48","author":"Winter","year":"2017","journal-title":"Lab. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1055\/s-2007-994929","article-title":"Hereditary and acquired antithrombin deficiency","volume":"25","author":"Vinazzer","year":"1999","journal-title":"Semin. Thromb. Hemost."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2118","DOI":"10.1016\/j.bbamcr.2017.07.009","article-title":"The plasma contact system, a protease cascade at the nexus of inflammation, coagulation and immunity","volume":"1864","author":"Weidmann","year":"2017","journal-title":"Biochim. Biophys. Acta Mol. Cell Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/0009-8981(67)90104-0","article-title":"Alpha-1-antitrypsin, an inhibitor for thrombin and plasmin","volume":"17","author":"Gans","year":"1967","journal-title":"Clin. Chim. Acta"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"6119","DOI":"10.3748\/wjg.v16.i48.6119","article-title":"High prevalence of nonalcoholic fatty liver in patients with idiopathic venous thromboembolism","volume":"16","author":"Tufano","year":"2010","journal-title":"World J. Gastroenterol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Hu, P.A., Chen, C.H., Guo, B.C., Kou, Y.R., and Lee, T.S. (2020). Bromelain confers protection against the non-alcoholic fatty liver disease in male C57BL\/6 mice. Nutrients, 12.","DOI":"10.3390\/nu12051458"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"283","DOI":"10.3892\/br.2016.720","article-title":"Potential role of bromelain in clinical and therapeutic applications","volume":"5","author":"Rathnavelu","year":"2016","journal-title":"Biomed. Rep."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1177\/000331976902000103","article-title":"Oral bromelains as adjuncts in the treatment of acute thrombophlebitis","volume":"20","author":"Seligman","year":"1969","journal-title":"Angiology"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1016\/0306-9877(80)90134-6","article-title":"Fibrinolytic and antithrombotic action of bromelain may eliminate thrombosis in heart patients","volume":"6","author":"Felton","year":"1980","journal-title":"Med. Hypotheses"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1016\/S0021-9258(18)53998-3","article-title":"Complex-dependent inhibition of factor VIIa by antithrombin III and heparin","volume":"268","author":"Lawson","year":"1993","journal-title":"J. Biol. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1111\/j.1538-7836.2005.01679.x","article-title":"Fatal thrombosis of antithrombin-deficient mice is rescued differently in the heart and liver by intercrossing with low tissue factor mice","volume":"4","author":"Hayashi","year":"2006","journal-title":"J. Thromb. Haemost."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1042\/bj1590495","article-title":"Inactivation of human thrombin in the presence of human \u03b1-proteinase inhibitor","volume":"159","author":"Matheson","year":"1976","journal-title":"Biochem. J."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1016\/j.jaci.2013.04.006","article-title":"De novo homozygous mutation of the C1 inhibitor gene in a patient with hereditary angioedema","volume":"132","author":"Bafunno","year":"2013","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1097\/00001721-200106000-00005","article-title":"In vitro interaction of C1-inhibitor with thrombin","volume":"12","author":"Cugno","year":"2001","journal-title":"Blood Coagul. Fibrinolysis"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1007\/978-3-319-49674-0_13","article-title":"Fibrin Formation, Structure and Properties","volume":"82","author":"Weisel","year":"2017","journal-title":"Subcell. Biochem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/S0065-3233(05)70008-5","article-title":"Fibrinogen and fibrin","volume":"70","author":"Weisel","year":"2005","journal-title":"Adv. Protein Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/S0049-3848(99)00023-7","article-title":"Blood coagulation factor XIII: Structure and function","volume":"94","author":"Muszbek","year":"1999","journal-title":"Thromb. Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1172\/JCI114211","article-title":"Interaction between plasminogen activator inhibitor type 1 (PAI-1) bound to fibrin and either tissue-type plasminogen activator (t-PA) or urokinase-type plasminogen activator (u-PA). Binding of t-PA\/PAI-1 complexes to fibrin mediated by both the finger and the kringle-2 domain of t-PA","volume":"84","author":"Wagner","year":"1989","journal-title":"J. Clin. Investig."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1007\/BF01923166","article-title":"Effect of bromelain (Ananase) on human platelet aggregation","volume":"28","author":"Heinicke","year":"1972","journal-title":"Experientia"}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/14\/11\/2329\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:23:22Z","timestamp":1760138602000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/14\/11\/2329"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,1]]},"references-count":37,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2022,6]]}},"alternative-id":["nu14112329"],"URL":"https:\/\/doi.org\/10.3390\/nu14112329","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,6,1]]}}}