Hippasa agelenoides spider venom gland extract exhibits myotoxic , fibrino ( gen ) lytic and platelet agregation inhibitory properties

The present study explores the biochemical and pharmacological examination of H. agelenoides spider venom gland extract for the proteolytic activity and along with other pharmacological properties. The venom gland extract hydrolyzed casein, the proteolytic activity was further confirmed by the activity staining on casein zymogram. The venom gland extract showed two translucent activity bands on casein zymogram in the region of 16 kDa and 29 kDa suggesting the presence of two isoforms of proteases. However, both the activity bands were sensitive to PMSF but insensitive to EGTA, EDTA, 1, 10, phenanthroline and IAA, suggesting both isoforms are belongs to serine protease family. The venom gland extract was devoid of hemorrhage and cytotoxicity but caused edema and myotoxicity in experimental mice. The histopathology of transverse section of mice skeletal muscle treated with the venom gland extract showed extensive destruction of myocytes. Further, the histopathology of venom gland extract treated skin tissue revealed extensive damage of extra cellular matrix (ECM) of hypodermis with out causing any damage to dermis, epidermis, blood vessels and capillaries. The degradation of ECM components was further confirmed by in-vitro studies using ECM components such as collagen types-I and IV and fibronectin. The venom gland extract hydrolyzed specifically collagen type-I and fibronectin in a dose dependent manner, but it did not degrade gelatin and collagen type-IV. Interestingly, the venom gland extract caused procoagulation in the citrated human plasma. Further, the venom gland extract showed fibrino(geno)lytic activity. It readily and preferentially hydrolyzed the Aα-chain followed by Bβ-chain, but without affecting the γ-chain of human fibrinogen. However, it hydrolyzed all the subunits of partially cross-liked fibrin clot (α-polymer, α-chain, β-chain, and γ-γ dimers). In addition, the venom gland extract was also interfered in platelet function. It inhibited the collagen induced platelet aggregation in platelet rich plasma (PRP) and washed human platelets.