Exploring hemostatic and thrombolytic potential of heynein - A cysteine protease from Ervatamia heyneana latex

• Published in: Journal of ethnopharmacology 2017-03, Vol.199, p.316-322
• Main Authors: Uday, Priyanka, Maheshwari, M., Sharanappa, P., Nafeesa, Zohara, Kameshwar, Vivek Hamse, Priya, B.S., Nanjunda Swamy, S.
• Format: Article
• Language: English
• Subjects: Adult; Animals; Apocynaceae; Cysteine Proteases - isolation & purification; Cysteine Proteases - pharmacology; Cysteine Proteases - therapeutic use; Ervatamia heyneana (Wall.) T. Cooke; Fibrino(geno)lytic; Fibrinogen - metabolism; Fibrinolytic Agents - isolation & purification; Fibrinolytic Agents - pharmacology; Fibrinolytic Agents - therapeutic use; Hemorrhage - drug therapy; Hemorrhage - metabolism; Hemostasis; Hemostasis - drug effects; Hemostasis - physiology; Hemostatics - isolation & purification; Hemostatics - pharmacology; Hemostatics - therapeutic use; Heynein (cysteine protease); Humans; Latex - isolation & purification; Latex - pharmacology; Latex - therapeutic use; Male; Mice; Plant Bark; Plant Extracts - isolation & purification; Plant Extracts - pharmacology; Plant Extracts - therapeutic use; Pro–coagulant; Thrombosis - drug therapy; Thrombosis - metabolism; Young Adult
• ISSN: 0378-8741; 1872-7573
• DOI: 10.1016/j.jep.2016.12.047

The latex of Ervatamia heyneana (Wall.) T. Cooke plant has been used for wound healing and various skin diseases by Indian tribes and folklore. To validate the scientific basis of heynein – a key protease of Ervatamia heyneana, in hemostasis and wound healing process. The latex from E. heyneana was processed and subjected to two step purification. The purified heynein was assayed for proteolytic activity using casein as substrate and also attested by zymography. The inhibition studies confirmed the nature of heynein. Pure fibrinogen was used for fibrinogenolytic activity and citrated plasma was used for coagulant and fibrinolytic activities. The edema inducing action and hemorrhagic activity of heynein were assessed on mice model. The purified heynein exhibited proteolytic activity, which was confirmed by caseinolytic assay and zymography. The inhibition studies confirmed heynein to be a cysteine protease. Heynein showed complete hydrolysis of all the three subunits of human fibrinogen (Aα, Bβ, γ). It exhibited strong pro-coagulant activity by reducing plasma clotting time from 248 to 39s at 40µg concentration. Heynein cleaved α polymer subunit in fibrin clot and did not induce edema and hemorrhage in mice models. The non-hemorrhagic nature was supported with histopathological studies of skin samples. Heynein displays strong pro-coagulant action associated with fibrin(ogen)olytic activity. This provides basis for the observed pharmacological action of Ervatamia heyneana and thereby justifies its use in folk medicine. [Display omitted]