Cytotoxic Effect of Iranian Vipera lebetina Snake Venom on HUVEC Cells

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Abstract

Objective: Envenomation by heamotoxic snakes constituted a critical health occurrence in the world. Bleeding is the most sever consequence following snake bite with viperid and crothalid snakes. It is believed that the degradation of vascular membrane caused hemorrhage; in contrast, some suggested that direct cytotoxicity has role in endothelial cell disturbances. This study was carried out to evaluate the direct toxicity effect of V. lebetina crude venom on Human Umbilical Vein Endothelial Cells (HUVECs). Methods: The effect of V. lebetina snake venom on HUVECs growth inhibition was determined by MTT assay and neutral red uptake assay. The integrity of cell membrane through LDH release was measured with the Cytotoxicity Detection Kit. Morphological changes of endothelial cells were also evaluated using a phase contrast microscope. Result: In MTT assay, crude venom showed a cytotoxic effect on endothelial cells which was confirmed by the effect observed with neutral red assay. Also, crude venom caused changes in the integrity of cell membrane by LDH release. The morphological alterations enhanced in high concentration results in total cells number reduced. Conclusion: V. lebetina venom showed potential direct cytotoxic effects on human endothelial cells in a manner of concentration- dependent inhibition.

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Detection of necrosis by release of lactate dehydrogenase activity.

Francis Chan, Kenta Moriwaki, Maria De Rosa (2013)

Apoptosis and necrosis are two major forms of cell death observed in normal and disease pathologies. Although there are many assays for detection of apoptosis, relatively few assays are available for measuring necrosis. A key signature for necrotic cells is the permeabilization of the plasma membrane. This event can be quantified in tissue culture settings by measuring the release of the intracellular enzyme lactate dehydrogenase (LDH). When combined with other methods, measuring LDH release is a useful method for the detection of necrosis. In this chapter, we describe the step-by-step procedure for detection of LDH release from necrotic cells using a microtiter plate-based colorimetric absorbance assay.

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Snake venom metalloproteinases: their role in the pathogenesis of local tissue damage.

J Gutiérrez, A Rucavado (2000)

The biochemical characteristics of hemorrhagic metalloproteinases isolated from snake venoms are reviewed, together with their role in the pathogenesis of the local tissue damage characteristic of crotaline and viperine snake envenomations. Venom metalloproteinases differ in their domain structure. Some enzymes comprise only the metalloproteinase domain, others have disintegrin-like and high cysteine domains and others present, besides these domains, an additional lectin-like subunit. All of them are zinc-dependent enzymes with highly similar zinc binding environments. Some metalloproteinases induce hemorrhage by directly affecting mostly capillary blood vessels. It is suggested that hemorrhagic enzymes cleave, in a highly selective fashion, key peptide bonds of basement membrane components, thereby affecting the interaction between basement membrane and endothelial cells. As a consequence, these cells undergo a series of morphological and functional alterations in vivo, probably associated with biophysical hemodynamic factors such as tangential fluid shear stress. Eventually, gaps are formed in endothelial cells through which extravasation occurs. In addition to hemorrhage, venom metalloproteinases induce skeletal muscle damage, myonecrosis, which seems to be secondary to the ischemia that ensues in muscle tissue as a consequence of bleeding and reduced perfusion. Microvessel disruption by metalloproteinases also impairs skeletal muscle regeneration, being therefore responsible of fibrosis and permanent tissue loss after snakebites. Moreover, venom metalloproteinases participate in the degradation of extracellular matrix components and play a relevant role in the prominent local inflammatory response that characterizes snakebite envenomations, since they induce edema, activate endogenous matrix metalloproteinases (MMPs) and are capable of releasing TNF-alpha from its membrane-bound precursor. Owing to their protagonic role in the pathogenesis of local tissue damage, snake venom metalloproteinases constitute relevant targets for natural and synthetic inhibitors which may complement antivenoms in the neutralization of these effects.

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Snake venom components and their applications in biomedicine.

D Koh, A Armugam, K Jeyaseelan (2006)

Snake envenomation is a socio-medical problem of considerable magnitude. About 2.5 million people are bitten by snakes annually, more than 100,000 fatally. However, although bites can be deadly, snake venom is a natural biological resource that contains several components of potential therapeutic value. Venom has been used in the treatment of a variety of pathophysiological conditions in Ayurveda, homeopathy and folk medicine. With the advent of biotechnology, the efficacy of such treatments has been substantiated by purifying components of venom and delineating their therapeutic properties. This review will focus on certain snake venom components and their applications in health and disease.

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Author and article information

Journal

Journal ID (nlm-ta): Iran J Pharm Res

Journal ID (iso-abbrev): Iran J Pharm Res

Journal ID (publisher-id): IJPR

Title: Iranian Journal of Pharmaceutical Research : IJPR

Publisher: Shaheed Beheshti University of Medical Sciences (Tehran, Iran )

ISSN (Print): 1735-0328

ISSN (Electronic): 1726-6890

Publication date (Print): Winter 2015

Volume: 14

Issue: Suppl

Pages: 109-114

Affiliations

[a ] Toxicology and Pharmacology Department, School of Pharmacy, Medical Science University of Shahid Beheshti, Tehran, Iran.

[b ] Pharmacology Department, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

[c ] Venomous Animals and Antivenom Production Department, Razi Vaccine and Serum Research Institute, Karaj, Iran.

[d ] Toxicology Department, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

Author notes

[* ]Corresponding author: E.mail: vatanpour.hossein@gmail.com

Article

Publisher ID: ijpr-14-109

PMC ID: 4499433

SO-VID: 44c52449-ae05-436a-af66-64b6c5ede657

License:

This is an Open Access article distributed under the terms of the Creative Commons Attribution License, ( http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Cytotoxic Effect of Iranian Vipera lebetina Snake Venom on HUVEC Cells

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Gamma Delta T cells

Most cited references 36

Detection of necrosis by release of lactate dehydrogenase activity.

Snake venom metalloproteinases: their role in the pathogenesis of local tissue damage.

Snake venom components and their applications in biomedicine.

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