{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T16:07:47Z","timestamp":1791216467438,"version":"4.1.0"},"reference-count":54,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2023,11,26]],"date-time":"2023-11-26T00:00:00Z","timestamp":1700956800000},"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>While fibrinolytic enzymes and thrombolytic agents offer assistance in treating cardiovascular diseases, the existing options are associated with a range of adverse effects. In our previous research, we successfully identified ficin, a naturally occurring cysteine protease that possesses unique fibrin and fibrinogenolytic enzymes, making it suitable for both preventing and treating cardiovascular disorders linked to thrombosis. Papain is a prominent cysteine protease derived from the latex of Carica papaya. The potential role of papain in preventing fibrino(geno)lytic, anticoagulant, and antithrombotic activities has not yet been investigated. Therefore, we examined how papain influences fibrinogen and the process of blood coagulation. Papain is highly stable at pH 4\u201311 and 37\u201360 \u00b0C via azocasein assay. In addition, SDS gel separation electrophoresis, zymography, and fibrin plate assays were used to determine fibrinogen and fibrinolysis activity. Papain has a molecular weight of around 37 kDa, and is highly effective in degrading fibrin, with a molecular weight of over 75 kDa. Furthermore, papain-based hemostatic performance was confirmed in blood coagulation tests, a blood clot lysis assay, and a \u03ba-carrageenan rat tail thrombosis model, highlighting its strong efficacy in blood coagulation. Papain shows dose-dependent blood clot lysis activity, cleaves fibrinogen chains of A\u03b1, B\u03b2, and \u03b3-bands, and significantly extends prothrombin time (PT) and activated partial thromboplastin time (aPTT). Moreover, the mean length of the infarcted regions in the tails of Sprague\u2013Dawley rats with \u03ba-carrageenan was shorter in rats administered 10 U\/kg of papain than in streptokinase-treated rats. Thus, papain, a cysteine protease, has distinct fibrin and fibrinogenolytic properties, suggesting its potential for preventing or treating cardiovascular issues and thrombosis-related diseases.<\/jats:p>","DOI":"10.3390\/ijms242316770","type":"journal-article","created":{"date-parts":[[2023,11,27]],"date-time":"2023-11-27T03:32:03Z","timestamp":1701055923000},"page":"16770","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Unveiling the Potent Fibrino(geno)lytic, Anticoagulant, and Antithrombotic Effects of Papain, a Cysteine Protease from Carica papaya Latex Using \u03ba-Carrageenan Rat Tail Thrombosis Model"],"prefix":"10.3390","volume":"24","author":[{"given":"Hye Ryeon","family":"Yang","sequence":"first","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Most Nusrat","family":"Zahan","sequence":"additional","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yewon","family":"Yoon","sequence":"additional","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyuri","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7228-3340","authenticated-orcid":false,"given":"Du Hyeon","family":"Hwang","sequence":"additional","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6874-6787","authenticated-orcid":false,"given":"Woo Hyun","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea"},{"name":"Institute of Animal Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Il Rae","family":"Rho","sequence":"additional","affiliation":[{"name":"Institutes of Agriculture and Life Science, Gyeongsang National University, Jinju 52828, Republic of 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, Republic of Korea"},{"name":"Institute of Animal Medicine, Gyeongsang National University, Jinju 52828, Republic of 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, Republic of Korea"},{"name":"Institute of Animal Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,11,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"234","DOI":"10.30574\/gscbps.2021.14.3.0073","article-title":"Morphology, Phytochemistry and Pharmacological Aspects of Carica Papaya, an Review","volume":"14","author":"Wadekar","year":"2021","journal-title":"GSC Biol. Pharm. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"735","DOI":"10.1007\/s40199-020-00348-7","article-title":"Therapeutic Application of Carica Papaya Leaf Extract in the Management of Human Diseases","volume":"28","author":"Singh","year":"2020","journal-title":"DARU J. Pharm. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Gracz-bernaciak, J., Mazur, O., and Nawrot, R. (2021). Functional Studies of Plant Latex as a Rich Source of Bioactive Compounds: Focus on Proteins and Alkaloids. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222212427"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"M\u00f3ty\u00e1n, J.A., T\u00f3th, F., and T\u0151zs\u00e9r, J. (2013). Research Applications of Proteolytic Enzymes in Molecular Biology. Biomolecules, 3.","DOI":"10.3390\/biom3040923"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Razzaq, A., Shamsi, S., Ali, A., Ali, Q., Sajjad, M., Malik, A., and Ashraf, M. (2019). Microbial Proteases Applications. Front. Bioeng. Biotechnol., 7.","DOI":"10.3389\/fbioe.2019.00110"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"596","DOI":"10.3923\/pjbs.2010.596.603","article-title":"Efficacy of Papain-Based Wound Cleanser in Promoting Wound Regeneration","volume":"13","author":"Ajlia","year":"2010","journal-title":"Pak. J. Biol. Sci."},{"key":"ref_7","first-page":"876426","article-title":"Natural Products for Antithrombosis","volume":"2015","author":"Chen","year":"2015","journal-title":"Evid.-Based Complement. Altern. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1186\/s43162-021-00090-9","article-title":"Current Update on Herbal Sources of Antithrombotic Activity\u2014A Comprehensive Review","volume":"34","author":"Subramani","year":"2022","journal-title":"Egypt. J. Intern. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1007\/s13167-022-00293-2","article-title":"Antithrombotic and Antiplatelet Effects of Plant-Derived Compounds: A Great Utility Potential for Primary, Secondary, and Tertiary Care in the Framework of 3P Medicine","volume":"13","author":"Kubatka","year":"2022","journal-title":"EPMA J."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1038\/nrcardio.2017.206","article-title":"Current and Future Antiplatelet Therapies: Emphasis on Preserving Haemostasis","volume":"15","author":"McFadyen","year":"2018","journal-title":"Nat. Rev. Cardiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2153","DOI":"10.1002\/med.21579","article-title":"Bioactive Peptides and Proteins as Alternative Antiplatelet Drugs","volume":"39","author":"Rengasamy","year":"2019","journal-title":"Med. Res. Rev."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s12959-018-0170-8","article-title":"Human Blood Platelets and Viruses: Defense Mechanism and Role in the Removal of Viral Pathogens","volume":"16","author":"Seyoum","year":"2018","journal-title":"Thromb. J."},{"key":"ref_13","first-page":"319","article-title":"Mechanism Action of Platelets and Crucial Blood Coagulation Pathways in Hemostasis","volume":"11","author":"Periayah","year":"2017","journal-title":"Int. J. Hematol. Stem Cell Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1111\/j.1365-2796.2004.01444.x","article-title":"Blood Coagulation and Its Regulation by Anticoagulant Pathways: Genetic Pathogenesis of Bleeding and Thrombotic Diseases","volume":"257","year":"2005","journal-title":"J. Intern. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2918","DOI":"10.1111\/jth.15538","article-title":"Clinical Use of Thrombin Generation Assays","volume":"19","author":"Binder","year":"2021","journal-title":"J. Thromb. Haemost."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"41","DOI":"10.2165\/00003495-197612010-00002","article-title":"Antithrombotic Drugs: Part I","volume":"12","author":"Gallus","year":"1976","journal-title":"Drugs"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1577","DOI":"10.1111\/j.1538-7836.2005.01418.x","article-title":"Therapeutic Approaches in Arterial Thrombosis","volume":"3","author":"Phillips","year":"2005","journal-title":"J. Thromb. Haemost."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2142","DOI":"10.1016\/j.jacc.2020.07.070","article-title":"Antithrombotic Management of Venous Thromboembolism: JACC Focus Seminar","volume":"76","author":"Renner","year":"2020","journal-title":"J. Am. Coll. Cardiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"100615","DOI":"10.1016\/j.blre.2019.100615","article-title":"From Multi-Target Anticoagulants to DOACs, and Intrinsic Coagulation Factor Inhibitors","volume":"39","author":"Lin","year":"2020","journal-title":"Blood Rev."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.thromres.2009.03.015","article-title":"Platelet Reactivity on Aspirin, Clopidogrel and Abciximab in Patients with Acute Coronary Syndromes and Reduced Estimated Glomerular Filtration Rate","volume":"125","author":"Sitkiewicz","year":"2010","journal-title":"Thromb. Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1145","DOI":"10.1001\/archinte.163.10.1145","article-title":"Aspirin and Clopidogrel in Acute Coronary Syndromes: Therapeutic Insights From the CURE Study","volume":"163","author":"Jneid","year":"2003","journal-title":"Arch. Intern. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1111\/j.1747-4949.2012.00923.x","article-title":"Thrombolysis in the Developing World: Is There a Role for Streptokinase?","volume":"8","author":"Butcher","year":"2013","journal-title":"Int. J. Stroke"},{"key":"ref_23","unstructured":"Baig, M.U., and Bodle, J. (2022). StatPearls, StatPearls Publishing."},{"key":"ref_24","first-page":"274","article-title":"Adverse Reactions to Thrombolytic Agents: Implications for Coronary Reperfusion Following Myocardial Infarction","volume":"2","author":"Nazari","year":"1987","journal-title":"Med. Toxicol. Advers. Drug Exp."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Yang, H.R., Hwang, D.H., Prakash, R.L.M., Kim, J.H., Hong, I.H., Kim, S., Kim, E., and Kang, C. (2022). Exploring the Fibrin(Ogen)Olytic, Anticoagulant, and Antithrombotic Activities of Natural Cysteine Protease (Ficin) with the \u03ba-Carrageenan-Induced Rat Tail Thrombosis Model. Nutrients, 14.","DOI":"10.3390\/nu14173552"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"012034","DOI":"10.1088\/1755-1315\/196\/1\/012034","article-title":"Characteristics of Papain Soluble Collagen from Redbelly Yellowtail Fusilier (Caesio Cuning)","volume":"196","author":"Nurhayati","year":"2018","journal-title":"IOP Conf. Ser. Earth Environ. Sci."},{"key":"ref_27","unstructured":"Rawlings, N.D., and Salvesen, G. (2013). Handbook of Proteolytic Enzymes, Elsevier."},{"key":"ref_28","unstructured":"Grzonka, Z., Kasprzykowski, F., and Wiczk, W. (2007). Industrial Enzymes, Springer."},{"key":"ref_29","unstructured":"Singh, R., Singh, A., and Sachan, S. (2018). Enzymes in Food Biotechnology Production, Applications, and Future Prospects, Elsevier."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1007\/s13596-022-00670-2","article-title":"Traditional Herbal Medicine: Overview of Research Indexed in the Scopus Database","volume":"23","author":"Musa","year":"2022","journal-title":"Adv. Tradit. Med."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Kang, Y.M., Kang, H.A., Cominguez, D.C., Kim, S.H., and An, H.J. (2021). Papain Ameliorates Lipid Accumulation and Inflammation in High-Fat Diet-Induced Obesity Mice and 3t3-L1 Adipocytes via Ampk Activation. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22189885"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"99","DOI":"10.3844\/ajbbsp.2012.99.104","article-title":"Papain, a Plant Enzyme of Biological Importance: A Review","volume":"8","author":"Amri","year":"2012","journal-title":"Am. J. Biochem. Biotechnol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2932","DOI":"10.1002\/jsfa.10320","article-title":"Effects of Drying on Total Polyphenols Content and Antioxidant Properties of Carica Papaya Leaves","volume":"100","author":"Yap","year":"2020","journal-title":"J. Sci. Food Agric."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"105369","DOI":"10.1016\/j.arabjc.2023.105369","article-title":"Therapeutic Benefits of Carica Papaya: A Review on Its Pharmacological Activities and Characterization of Papain","volume":"17","author":"Otunba","year":"2024","journal-title":"Arab. J. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1085\/jgp.20.4.543","article-title":"Studies in Blood Coagulation v. the Coagulation of Blood by Proteolytic Enzymes (Trypsin, Papain)","volume":"20","author":"Eagle","year":"1937","journal-title":"J. Gen. Physiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1079","DOI":"10.1039\/D2TB02220F","article-title":"A Cysteine Enzyme Hemostat for Efficient Heparin-Tolerant Blood Coagulation","volume":"11","author":"Lin","year":"2023","journal-title":"J. Mater. Chem. B"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Fei, X., Yuan, W., Zhao, Y., Wang, H., Bai, S., and Huang, Q. (2018). Papain Ameliorates the MPAs Formation-Mediated Activation of Monocytes by Inhibiting Cox-2 Expression via Regulating the MAPKs and PI3K\/Akt Signal Pathway. Biomed Res. Int., 2018.","DOI":"10.1155\/2018\/3632084"},{"key":"ref_38","first-page":"7125162","article-title":"A Review of Antiplatelet Activity of Traditional Medicinal Herbs on Integrative Medicine Studies","volume":"2019","author":"Kim","year":"2019","journal-title":"Evid.-Based Complement. Altern. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2052","DOI":"10.1161\/01.CIR.94.9.2052","article-title":"Impaired Fibrinolysis and the Risk for Coronary Heart Disease","volume":"94","author":"Lijnen","year":"1996","journal-title":"Circulation"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"461233","DOI":"10.3389\/fimmu.2019.01348","article-title":"Fibrinolysis and Inflammation in Venous Thrombus Resolution","volume":"10","author":"Mukhopadhyay","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"932","DOI":"10.1111\/j.1538-7836.2006.01861.x","article-title":"Fibrin Structure and Wound Healing","volume":"4","author":"Laurens","year":"2006","journal-title":"J. Thromb. Haemost."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.blre.2014.09.003","article-title":"Fibrinolysis and the Control of Blood Coagulation","volume":"29","author":"Chapin","year":"2015","journal-title":"Blood Rev."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"670","DOI":"10.1055\/s-0028-1104546","article-title":"Standardization and Clinical Utility of Thrombin-Generation Assays","volume":"34","author":"Berntorp","year":"2008","journal-title":"Semin. Thromb. Hemost."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"5560560","DOI":"10.1155\/2023\/5560560","article-title":"Coagulation Dysfunction in Patients with Liver Cirrhosis and Splenomegaly and Its Countermeasures: A Retrospective Study of 1522 Patients","volume":"2023","author":"Lv","year":"2023","journal-title":"Dis. Markers"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1336","DOI":"10.2174\/1389201023666220104143113","article-title":"Fibrinolytic Enzyme\u2014An Overview","volume":"23","author":"Rengaswamy","year":"2022","journal-title":"Curr. Pharm. Biotechnol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"500","DOI":"10.4236\/wjcd.2020.107050","article-title":"Study of Adverse Events of Streptokinase Therapy in Patients with Acute ST Elevation Myocardial Infarction","volume":"10","author":"Rahman","year":"2020","journal-title":"World J. Cardiovasc. Dis."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.molimm.2019.02.020","article-title":"Molecular Mechanism of Action Responsible for Carrageenan-Induced Inflammatory Response","volume":"109","author":"Myers","year":"2019","journal-title":"Mol. Immunol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1185","DOI":"10.1016\/j.biocel.2007.12.009","article-title":"Proteolytic Processing of Chemokines: Implications in Physiological and Pathological Conditions","volume":"40","author":"Wolf","year":"2008","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Repnik, U., Starr, A.E., Overall, C.M., and Turk, B. (2015). Cysteine Cathepsins Activate ELR Chemokines and Inactivate Non-ELR Chemokines. J. Biol. Chem., 290.","DOI":"10.1074\/jbc.M115.638395"},{"key":"ref_50","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_51","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_52","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_53","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_54","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/S0305-0491(99)00161-3","article-title":"Purification and Characterization of a Fibrinolytic Enzyme and Identification of Fibrinogen Clotting Enzyme in a Marine Green Alga, Codium Divaricatum","volume":"125","author":"Matsubara","year":"2000","journal-title":"Comp. Biochem. Physiol. Part B Biochem. Mol. Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/23\/16770\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:30:37Z","timestamp":1760131837000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/23\/16770"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,11,26]]},"references-count":54,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2023,12]]}},"alternative-id":["ijms242316770"],"URL":"https:\/\/doi.org\/10.3390\/ijms242316770","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,11,26]]}}}