Objective: The current study assesses the fibrinogen and fibrin clot hydrolyzing activities of aqueous seed extract of Jackfruit (AqSEJ). Methods: The protein banding pattern of AqSEJ (100 μg) was analyzed on SDS-PAGE. The proteolytic activity of AqSEJ was confirmed by spectrophotometer and zymography experiments. Fibrinogen, fibrin and plasma protein hydrolyzing activities of AqSEJ were analyzed on SDS-PAGE under reduced conditions. Plasminogen activation and indirect hemolytic activities was analyzed using spectrophotometer. The non-toxic property of AqSEJ was tested by edema, hemorrhage in experimental mice. Results: AqSEJ exhibited proteolytic activity and the specific activity was found to be 1.04 units/mg/min. Furthermore, AqSEJ non-specifically hydrolyzed Aα, followed by Bβ and γ chains of human fibrinogen and specifically hydrolyzed α polymer and α chain of partially cross linked human fibrin clot without affecting β chain and γ-γ dimer even up to the tested dose of 30 μg for the incubation period of 8 hours. Importantly, AqSEJ did not hydrolyze other plasma proteins and devoid of plasminogen activation property. The proteolytic activity of AqSEJ was completely neutralized by PMSF and IAA, while EDTA, EGTA, 1,10-Phenanthroline did not, suggesting the presence of serine and cysteine family proteases. Moreover, AqSEJ did not cause edema and hemorrhage in experimental mice up to the tested dose of 200 μg and non-toxic to RBC cells. Conclusion: AqSEJ hydrolyzes fibrinogen and fibrin clot and non-toxic in nature. Hence, this work showcases the potential applications of Jack fruit seed proteases in the treatment of thrombotic disorders.

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The present work demonstrates on the comparative studies on Aqueous and Ethanol Extracts of Momordica charantia (bitter gourd) Seed on plasma re-calcification time and platelet function. Anticoagulant effect of Aqueous and Ethanol Extracts of Momordica charantia was screened for tests such as plasma recalcification time, platelet aggregation studies and non-toxic property was tested by edema, hemorrhage and indirect hemolytic activities. Aqueous Extract seed (AEMCS) showed strong anticoagulant effect by enhancing the plasma clotting time from control 146s to 432s, while, Ethanol Extract of Momordica charantia Seed (EEMCS) showed weak anticoagulant effect as it enhanced the clotting time from control 146s to 160s. Furthermore, both extracts were analyzed for agonists such as, Collagen, Adenosine diphosphate (ADP) and epinephrine induced platelet aggregation. Curiously, both the extracts were inhibited agonist induced platelet aggregation of Platelet rich plasma (PRP). The percentage of platelet aggregation inhibition for AEMCS was found to be 65%, 50% and 85% respectively for collagen, ADP and epinephrine induced platelet aggregation. However, 19%, 91% and 97% platelet aggregation inhibition for EEMCS on said agonists. In addition, AEMCS and EEMCS both devoid of RBC lysis, edema and hemorrhagic properties, suggesting its nontoxic nature. In conclusion; both aqueous and ethanol extract of Momordica charantia found to exhibit anticoagulant and antiplatelet activities; while, AEMCS showed strong anticoagulant potential with moderate antiplatelet efficiency. On the other hand, EEMCS exhibited strong antiplatelet activity but poor anticoagulation. Hence, it could be promising agent in the management of thrombotic disorders.