{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T16:08:15Z","timestamp":1791216495852,"version":"4.1.0"},"reference-count":32,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2022,8,29]],"date-time":"2022-08-29T00:00:00Z","timestamp":1661731200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Although fibrinolytic enzymes and thrombolytic agents help in cardiovascular disease treatment, those currently available have several side effects. This warrants the search for safer alternatives. Several natural cysteine protease preparations are used in traditional medicine to improve platelet aggregation and thrombosis-related diseases. Hence, this study aimed to investigate the effect of ficin, a natural cysteine protease, on fibrin(ogen) and blood coagulation. The optimal pH (pH 7) and temperature (37 \u00b0C) for proteolytic activity were determined using the azocasein method. Fibrinogen action and fibrinolytic activity were measured both electrophoretically and by the fibrin plate assay. The effect of ficin on blood coagulation was studied by conventional coagulation tests: prothrombin time (PT), activated partial thromboplastin time (aPTT), blood clot lysis assay, and the \u03ba-carrageenan thrombosis model. The A\u03b1, B\u03b2, and \u03b3 bands of fibrinogen are readily cleaved by ficin, and we also observed a significant increase in PT and aPTT. Further, the mean length of the infarcted regions in the tails of Sprague\u2013Dawley rats was shorter in rats administered 10 U\/mL of ficin than in control rats. These findings suggest that natural cysteine protease, ficin contains novel fibrin and fibrinogenolytic enzymes and can be used for preventing and\/or treating thrombosis-associated cardiovascular disorders.<\/jats:p>","DOI":"10.3390\/nu14173552","type":"journal-article","created":{"date-parts":[[2022,8,29]],"date-time":"2022-08-29T22:53:42Z","timestamp":1661813622000},"page":"3552","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Exploring the Fibrin(ogen)olytic, Anticoagulant, and Antithrombotic Activities of Natural Cysteine Protease (Ficin) with the \u03ba-Carrageenan-Induced Rat Tail Thrombosis Model"],"prefix":"10.3390","volume":"14","author":[{"given":"Hye Ryeon","family":"Yang","sequence":"first","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea"},{"name":"Institute of Animal Medicine, Gyeongsang National University, Jinju 52828, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Du Hyeon","family":"Hwang","sequence":"additional","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea"},{"name":"Institute of Animal Medicine, Gyeongsang National University, Jinju 52828, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramachandran Loganathan Mohan","family":"Prakash","sequence":"additional","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea"},{"name":"Institute of Animal Medicine, Gyeongsang National University, Jinju 52828, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jong-Hyun","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea"},{"name":"Institute of Animal Medicine, Gyeongsang National University, Jinju 52828, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Il-Hwa","family":"Hong","sequence":"additional","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea"},{"name":"Institute of Animal Medicine, Gyeongsang National University, Jinju 52828, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5795-2019","authenticated-orcid":false,"given":"Suk","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea"},{"name":"Institute of Animal Medicine, Gyeongsang National University, Jinju 52828, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3356-3072","authenticated-orcid":false,"given":"Euikyung","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea"},{"name":"Institute of Animal Medicine, Gyeongsang National University, Jinju 52828, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5114-5096","authenticated-orcid":false,"given":"Changkeun","family":"Kang","sequence":"additional","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea"},{"name":"Institute of Animal Medicine, Gyeongsang National University, Jinju 52828, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,29]]},"reference":[{"key":"ref_1","first-page":"8","article-title":"Pharmacognostic and Phytochemical Properties of Ficus carica Linn\u2014An Overview","volume":"3","author":"Joseph","year":"2011","journal-title":"Int. J. Pharmtech Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.toxicon.2005.03.012","article-title":"Procoagulant Activity of Calotropis Gigantea Latex Associated with Fibrin (Ogen) Olytic Activity","volume":"46","author":"Rajesh","year":"2005","journal-title":"Toxicon"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"870","DOI":"10.1016\/j.phytochem.2006.02.020","article-title":"Purification and Characterization of a Serine Protease from Cucumis trigonus Roxburghi","volume":"67","author":"Kim","year":"2006","journal-title":"Phytochemistry"},{"key":"ref_4","first-page":"339","article-title":"Botanical Briefs: The Fig-Ficus carica L.","volume":"69","author":"McGovern","year":"2002","journal-title":"Cutis"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1046\/j.0014-2956.2002.02620.x","article-title":"Characterization of a Partially Folded Intermediate of Stem Bromelain at Low pH","volume":"269","author":"Haq","year":"2002","journal-title":"Eur. J. Biochem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1057","DOI":"10.1016\/S0031-9422(02)00676-3","article-title":"Procerain, a Stable Cysteine Protease from the Latex of Calotropis Procera","volume":"62","author":"Dubey","year":"2003","journal-title":"Phytochemistry"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e00492","DOI":"10.1016\/j.btre.2020.e00492","article-title":"A Contradictory Action of Procoagulant Ficin by a Fibrinolytic Serine Protease from Egyptian Ficus carica Latex","volume":"27","author":"Hamed","year":"2020","journal-title":"Biotechnol. Rep."},{"key":"ref_8","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. Fibrinol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"568","DOI":"10.2119\/molmed.2010.00146","article-title":"Novel Aspects of Fibrin (Ogen) Fragments during Inflammation","volume":"17","author":"Jennewein","year":"2011","journal-title":"Mol. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"680397","DOI":"10.3389\/fmolb.2021.680397","article-title":"Role of Fibrinolytic Enzymes in Anti-Thrombosis Therapy","volume":"8","author":"Altaf","year":"2021","journal-title":"Front. Mol. Biosci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1042","DOI":"10.1046\/j.1365-2141.2002.03954.x","article-title":"Activation and Inactivation of Human Factor X by Proteases Derived from Ficus carica","volume":"119","author":"Richter","year":"2002","journal-title":"Br. J. Haematol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"702","DOI":"10.3736\/jcim20110702","article-title":"A Review of the use of Bromelain in Cardiovascular Diseases","volume":"9","author":"Ley","year":"2011","journal-title":"J. Chin. Integr. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1038\/227680a0","article-title":"Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4","volume":"227","author":"Laemmli","year":"1970","journal-title":"Nature"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2715","DOI":"10.1099\/00221287-140-10-2715","article-title":"The Nematophagous Fungus Verticillium chlamydosporium Produces a Chymoelastase-Like Protease which Hydrolyses Host Nematode Proteins in Situ","volume":"140","author":"Segers","year":"1994","journal-title":"Microbiology"},{"key":"ref_15","first-page":"2151","article-title":"Purification and Biochemical Characterization of a Novel Intestinal Protease from Scorpaena notata","volume":"20","author":"Aissaoui","year":"2017","journal-title":"Int. J. Food Prop."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1016\/0003-9861(52)90121-5","article-title":"The Fibrin Plate Method for Estimating Fibrinolytic Activity","volume":"40","author":"Astrup","year":"1952","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"S0003","DOI":"10.1006\/abio.1998.2816","article-title":"Fibrin Zymography: A Direct Analysis of Fibrinolytic Enzymes on Gels","volume":"263","author":"Kim","year":"1998","journal-title":"Anal. Biochem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/S0305-0491(97)00303-9","article-title":"Purification and Characterization of Two Fibrinolytic Enzymes from a Marine Green Alga, Codium intricatum","volume":"119","author":"Matsubara","year":"1998","journal-title":"Comp. Biochem. Physiol. Part B Biochem. Mol. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1186\/1477-9560-4-14","article-title":"Development of an in Vitro Model to Study Clot Lysis Activity of Thrombolytic Drugs","volume":"4","author":"Prasad","year":"2006","journal-title":"Thromb. J."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1718","DOI":"10.1016\/j.phytochem.2011.05.009","article-title":"Selective and Reversible Thiol-Pegylation, an Effective Approach for Purification and Characterization of Five Fully Active Ficin (Iso) Forms from Ficus carica Latex","volume":"72","author":"Azarkan","year":"2011","journal-title":"Phytochemistry"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1760","DOI":"10.1016\/j.biochi.2010.09.002","article-title":"Structural and Functional Comparison of Proteolytic Enzymes from Plant Latex and Snake Venoms","volume":"92","author":"Costa","year":"2010","journal-title":"Biochimie"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1111\/j.1538-7933.2004.00645.x","article-title":"Tissue-type Plasminogen Activator: A Historical Perspective and Personal Account","volume":"2","author":"Collen","year":"2004","journal-title":"J. Thromb. Haemost."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2482","DOI":"10.1128\/aem.62.7.2482-2488.1996","article-title":"Purification and Characterization of a Fibrinolytic Enzyme Produced from Bacillus Sp. Strain CK 11-4 Screened from Chungkook-Jang","volume":"62","author":"Kim","year":"1996","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1007\/s00253-005-0159-7","article-title":"Microbial Fibrinolytic Enzymes: An Overview of Source, Production, Properties, and Thrombolytic Activity in Vivo","volume":"69","author":"Peng","year":"2005","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"914","DOI":"10.1038\/nature06797","article-title":"Triggers, Targets and Treatments for Thrombosis","volume":"451","author":"Mackman","year":"2008","journal-title":"Nature"},{"key":"ref_26","first-page":"635","article-title":"Purification and Characterization of a Novel Fibrinolytic Enzyme by Candida guilliermondii Grown on Sunflower Oil Cake","volume":"8","author":"Rashad","year":"2012","journal-title":"J. Appl. Sci. Res"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"734","DOI":"10.2460\/ajvr.2001.62.734","article-title":"Effect of Storage Conditions on Hemostatic Parameters of Canine Plasma obtained for Transfusion","volume":"62","author":"Iazbik","year":"2001","journal-title":"Am. J. Vet. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1016\/j.cll.2009.05.002","article-title":"Prothrombin Time and Partial Thromboplastin Time Assay Considerations","volume":"29","author":"Ng","year":"2009","journal-title":"Clin. Lab. Med."},{"key":"ref_29","unstructured":"Kitchen, S., McCraw, A., and Echenagucia, M. (2010). Diagnosis of Hemophilia and Other Bleeding Disorders, World Federation of Hemophilia."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"444","DOI":"10.1007\/s11239-009-0333-3","article-title":"Thrombolytic Effect of Subtilisin QK on Carrageenan Induced Thrombosis Model in Mice","volume":"28","author":"Yan","year":"2009","journal-title":"J. Thromb. Thrombolysis"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1016\/j.phrs.2009.04.010","article-title":"Improving Frequency of Thrombosis by Altering Blood Flow in the Carrageenan-Induced Rat Tail Thrombosis Model","volume":"60","author":"Hagimori","year":"2009","journal-title":"Pharmacol. Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1016\/j.thromres.2010.11.028","article-title":"Antithrombotic Effects of Ethanol Extract of Crataegus orientalis in the Carrageenan-Induced Mice Tail Thrombosis Model","volume":"127","author":"Arslan","year":"2011","journal-title":"Thromb. Res."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/14\/17\/3552\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:19:23Z","timestamp":1760141963000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/14\/17\/3552"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,8,29]]},"references-count":32,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2022,9]]}},"alternative-id":["nu14173552"],"URL":"https:\/\/doi.org\/10.3390\/nu14173552","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,8,29]]}}}