Anticoagulant, Antiplatelet and Fibrinogenolytic Activity of Serine Protease from Cicer arietinum Seed Extract

Author(s): Chethana Ramachandraiah, Sharathkumar MNandish, Jayanna Kengaiah, Chandramma Srinivas, Ashwini Shiviaha, Sujatha MHanume gowda and Devaraja Sannaningaiah

Abstract: The present investigation aims to examine the role of anticoagulant, antiplatelet and fibrino(geno)lytic effect of Cicer arietinum Seed Extract (CASE). CASE showed similar protein banding pattern from 200kDa to 14kDa in both reduced and non-reduced conditions suggesting the presence of monomeric proteins. CASE showed proteolytic activity with the specific activity of 0.1units/mg/min. CASE delayed the clot formation process of both Platelet Rich Plasma (PRP) and Platelet Poor Plasma (PPP), from the control 210 sec to 410 sec and 300 sec to 600 sec respectively. CASE found to alter the clotting time of Activated Partial Thromboplastin Time but not Prothrombin Time suggesting, triggered anticoagulant effect by CASE could be due to its inference in intrinsic pathway of coagulation cascade. CASE hydrolyzed Aα & Bβ chains without disrupting γ chain of human fibrinogen. However, it hydrolyzed all the chains of human fibrin clot suggesting its limited proteolysis. The identified said activities were completely abolished by PMSF, a serine protease but not IAA, EDTA and 1, 10, Phenanthroline. Revealed the role of serine proteolytic enzyme. Furthermore, CASE inhibited platelet aggregation induced by agonists ADP and epinephrine in PRP. While, thrombin, collagen, arachidonic acid, ADP and epinephrine in washed platelets. The identified platelet aggregation inhibition was found to be 79%and 82% for PRP and 80%,71%,70%,67% & 58% respectively for washed platelets. CASE was non-hemorrhagic, non-edematic and non-hemolytic in nature.

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