Department of Biotechnology, JSS Science and Technology University, JSS Technical Institutions Campus, Mysuru 570 006, Karnataka, India.
Department of Studies in Biosciences, University of Mysore, Hemagangothri, Hassan, Karnataka 570006, India.
J Ethnopharmacol. 2017 Mar 6;199:316-322. doi: 10.1016/j.jep.2016.12.047. Epub 2017 Jan 2.
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.
印度部落和民间传说一直使用 Ervatamia heyneana (Wall.) T. Cooke 植物的乳胶来治疗伤口愈合和各种皮肤病。
验证 heynein——Ervatamia heyneana 的关键蛋白酶，在止血和伤口愈合过程中的科学基础。
从 E. heyneana 中提取乳胶并进行两步纯化。使用酪蛋白作为底物对纯化的 heynein 进行蛋白酶活性测定，并通过同工酶进行验证。抑制研究证实了 heynein 的性质。纯纤维蛋白原用于纤维蛋白原水解活性，柠檬酸盐血浆用于凝血和纤维蛋白溶解活性。在小鼠模型中评估了 heynein 的致水肿作用和出血活性。
纯化的 heynein 表现出蛋白酶活性，这通过酪蛋白水解测定和同工酶得到证实。抑制研究证实 heynein 是一种半胱氨酸蛋白酶。heynein 完全水解了人纤维蛋白原（Aα、Bβ、γ）的所有三个亚基。它在 40μg 浓度下通过将血浆凝固时间从 248 秒缩短至 39 秒，表现出很强的促凝血活性。heynein 裂解纤维蛋白凝块中的α聚合物亚基，并且在小鼠模型中不会引起水肿和出血。非出血性质得到皮肤样本组织病理学研究的支持。
heynein 显示出强烈的促凝血作用，与纤维蛋白（原）水解活性相关。这为观察到的 Ervatamia heyneana 的药理学作用提供了依据，从而证明了其在民间医学中的应用。
