Unveiling the Potent Fibrino(geno)lytic, Anticoagulant, and Antithrombotic Effects of Papain, a Cysteine Protease from <i>Carica papaya</i> Latex Using κ-Carrageenan Rat Tail Thrombosis Model
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 fibr...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-11-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/23/16770 |
| _version_ | 1797400129484881920 |
|---|---|
| author | Hye Ryeon Yang Most Nusrat Zahan Yewon Yoon Kyuri Kim Du Hyeon Hwang Woo Hyun Kim Il Rae Rho Euikyung Kim Changkeun Kang |
| author_facet | Hye Ryeon Yang Most Nusrat Zahan Yewon Yoon Kyuri Kim Du Hyeon Hwang Woo Hyun Kim Il Rae Rho Euikyung Kim Changkeun Kang |
| author_sort | Hye Ryeon Yang |
| collection | DOAJ |
| description | 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 <i>Carica papaya</i>. 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 4–11 and 37–60 °C 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 κ-carrageenan rat tail thrombosis model, highlighting its strong efficacy in blood coagulation. Papain shows dose-dependent blood clot lysis activity, cleaves fibrinogen chains of Aα, Bβ, and γ-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–Dawley rats with κ-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. |
| first_indexed | 2024-03-09T01:50:06Z |
| format | Article |
| id | doaj.art-a1460f29eddc4be09a9ae3a080fc07a9 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T01:50:06Z |
| publishDate | 2023-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-a1460f29eddc4be09a9ae3a080fc07a92023-12-08T15:17:04ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-11-0124231677010.3390/ijms242316770Unveiling the Potent Fibrino(geno)lytic, Anticoagulant, and Antithrombotic Effects of Papain, a Cysteine Protease from <i>Carica papaya</i> Latex Using κ-Carrageenan Rat Tail Thrombosis ModelHye Ryeon Yang0Most Nusrat Zahan1Yewon Yoon2Kyuri Kim3Du Hyeon Hwang4Woo Hyun Kim5Il Rae Rho6Euikyung Kim7Changkeun Kang8Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of KoreaDepartment of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of KoreaDepartment of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of KoreaDepartment of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of KoreaDepartment of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of KoreaDepartment of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of KoreaInstitutes of Agriculture and Life Science, Gyeongsang National University, Jinju 52828, Republic of KoreaDepartment of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of KoreaDepartment of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of KoreaWhile 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 <i>Carica papaya</i>. 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 4–11 and 37–60 °C 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 κ-carrageenan rat tail thrombosis model, highlighting its strong efficacy in blood coagulation. Papain shows dose-dependent blood clot lysis activity, cleaves fibrinogen chains of Aα, Bβ, and γ-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–Dawley rats with κ-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.https://www.mdpi.com/1422-0067/24/23/16770papaincysteine proteaseblood coagulationthrombosisκ-carrageenan |
| spellingShingle | Hye Ryeon Yang Most Nusrat Zahan Yewon Yoon Kyuri Kim Du Hyeon Hwang Woo Hyun Kim Il Rae Rho Euikyung Kim Changkeun Kang Unveiling the Potent Fibrino(geno)lytic, Anticoagulant, and Antithrombotic Effects of Papain, a Cysteine Protease from <i>Carica papaya</i> Latex Using κ-Carrageenan Rat Tail Thrombosis Model International Journal of Molecular Sciences papain cysteine protease blood coagulation thrombosis κ-carrageenan |
| title | Unveiling the Potent Fibrino(geno)lytic, Anticoagulant, and Antithrombotic Effects of Papain, a Cysteine Protease from <i>Carica papaya</i> Latex Using κ-Carrageenan Rat Tail Thrombosis Model |
| title_full | Unveiling the Potent Fibrino(geno)lytic, Anticoagulant, and Antithrombotic Effects of Papain, a Cysteine Protease from <i>Carica papaya</i> Latex Using κ-Carrageenan Rat Tail Thrombosis Model |
| title_fullStr | Unveiling the Potent Fibrino(geno)lytic, Anticoagulant, and Antithrombotic Effects of Papain, a Cysteine Protease from <i>Carica papaya</i> Latex Using κ-Carrageenan Rat Tail Thrombosis Model |
| title_full_unstemmed | Unveiling the Potent Fibrino(geno)lytic, Anticoagulant, and Antithrombotic Effects of Papain, a Cysteine Protease from <i>Carica papaya</i> Latex Using κ-Carrageenan Rat Tail Thrombosis Model |
| title_short | Unveiling the Potent Fibrino(geno)lytic, Anticoagulant, and Antithrombotic Effects of Papain, a Cysteine Protease from <i>Carica papaya</i> Latex Using κ-Carrageenan Rat Tail Thrombosis Model |
| title_sort | unveiling the potent fibrino geno lytic anticoagulant and antithrombotic effects of papain a cysteine protease from i carica papaya i latex using κ carrageenan rat tail thrombosis model |
| topic | papain cysteine protease blood coagulation thrombosis κ-carrageenan |
| url | https://www.mdpi.com/1422-0067/24/23/16770 |
| work_keys_str_mv | AT hyeryeonyang unveilingthepotentfibrinogenolyticanticoagulantandantithromboticeffectsofpapainacysteineproteasefromicaricapapayailatexusingkcarrageenanrattailthrombosismodel AT mostnusratzahan unveilingthepotentfibrinogenolyticanticoagulantandantithromboticeffectsofpapainacysteineproteasefromicaricapapayailatexusingkcarrageenanrattailthrombosismodel AT yewonyoon unveilingthepotentfibrinogenolyticanticoagulantandantithromboticeffectsofpapainacysteineproteasefromicaricapapayailatexusingkcarrageenanrattailthrombosismodel AT kyurikim unveilingthepotentfibrinogenolyticanticoagulantandantithromboticeffectsofpapainacysteineproteasefromicaricapapayailatexusingkcarrageenanrattailthrombosismodel AT duhyeonhwang unveilingthepotentfibrinogenolyticanticoagulantandantithromboticeffectsofpapainacysteineproteasefromicaricapapayailatexusingkcarrageenanrattailthrombosismodel AT woohyunkim unveilingthepotentfibrinogenolyticanticoagulantandantithromboticeffectsofpapainacysteineproteasefromicaricapapayailatexusingkcarrageenanrattailthrombosismodel AT ilraerho unveilingthepotentfibrinogenolyticanticoagulantandantithromboticeffectsofpapainacysteineproteasefromicaricapapayailatexusingkcarrageenanrattailthrombosismodel AT euikyungkim unveilingthepotentfibrinogenolyticanticoagulantandantithromboticeffectsofpapainacysteineproteasefromicaricapapayailatexusingkcarrageenanrattailthrombosismodel AT changkeunkang unveilingthepotentfibrinogenolyticanticoagulantandantithromboticeffectsofpapainacysteineproteasefromicaricapapayailatexusingkcarrageenanrattailthrombosismodel |