Department of Basic Veterinary Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea.
Institute of Animal Medicine, Gyeongsang National University, Jinju 52828, Republic of Korea.
Int J Mol Sci. 2023 Nov 26;24(23):16770. doi: 10.3390/ijms242316770.
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 . 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.
虽然纤维蛋白溶解酶和溶栓剂在治疗心血管疾病方面提供了帮助，但现有的选择都伴随着一系列的不良反应。在我们之前的研究中，我们成功地鉴定了ficin，一种天然存在的半胱氨酸蛋白酶，具有独特的纤维蛋白和纤维蛋白原酶活性，使其适合预防和治疗与血栓形成相关的心血管疾病。木瓜蛋白酶是一种从橡胶树乳汁中提取的重要半胱氨酸蛋白酶。木瓜蛋白酶在防止纤维蛋白（原）溶解、抗凝和抗血栓形成方面的潜在作用尚未得到研究。因此，我们研究了木瓜蛋白酶如何影响纤维蛋白原和血液凝固过程。木瓜蛋白酶在 pH 值为 4-11 和 37-60°C 的情况下通过偶氮酪蛋白测定法高度稳定。此外，还使用 SDS 凝胶分离电泳、同工酶和纤维蛋白平板测定法来确定纤维蛋白原和纤维蛋白溶解活性。木瓜蛋白酶的分子量约为 37 kDa，对降解分子量超过 75 kDa 的纤维蛋白非常有效。此外，在血液凝固试验、血块溶解试验和κ-卡拉胶大鼠尾血栓模型中证实了基于木瓜蛋白酶的止血性能，突出了其在血液凝固方面的强大功效。木瓜蛋白酶表现出剂量依赖性的血块溶解活性，可切割纤维蛋白原 Aα、Bβ 和 γ 链，并显著延长凝血酶原时间 (PT) 和活化部分凝血活酶时间 (aPTT)。此外，κ-卡拉胶诱导的 Sprague-Dawley 大鼠尾部梗塞区域的平均长度在给予 10 U/kg 木瓜蛋白酶的大鼠中比给予链激酶治疗的大鼠更短。因此，木瓜蛋白酶作为一种半胱氨酸蛋白酶，具有独特的纤维蛋白和纤维蛋白原酶活性，表明其在预防或治疗心血管问题和与血栓形成相关的疾病方面具有潜力。
