title: Gene expression of inflammatory mediators induced by jararhagin on endothelial cells.
abstract: Snake venom metalloproteinases (SVMP) are abundant toxins in venoms of viper snakes and play a relevant role in the complex and multifactorial tissue damage characteristic of Viperidae envenoming. Jararhagin, a SVMP isolated from Bothrops jararaca venom, induces a fast onset hemorrhagic lesions acting directly on the capillary vessels, which are disrupted by toxin adhesion and degradation of extracellular matrix proteins like collagen IV. Jararhagin also triggers inflammatory response, where endothelial cells are activated, resulting in the enhanced rolling of circulating leukocytes, nitric oxide generation, prostacyclin production and pro-inflammatory cytokines release. Jararhagin also decreases endothelial cells viability inducing apoptosis (in vitro studies). In the present study we attempted to correlate the effect of sub-apoptotic doses of jararhagin on human umbilical vein endothelial cells (HUVECs) and gene expression of pro-inflammatory mediators, using microarray assay, real time PCR and detection of specific proteins on HUVEC surface or released in the medium. Jararhagin was effective in activate and up-regulate the gene expression of different mediators such as E-selectin, VCAM-1, IL-8, CD69, Ang-2 and MMP-10. Despite the increase in expression of genes coding for such molecules, jararhagin did not induce increased concentrations of E-selectin, VCAM-1 and IL-8 produced or released by endothelial cells. In conclusion, jararhagin is able to activate pro-inflammatory gene transcription on endothelial cells however this stimulus is not sufficient to result in the consequent expression of pro-inflammatory effectors molecules like E-selectin, VCAM-1 and IL-8. The time courses of these events, as well as the doses of jararhagin are important points to be addressed herein.
authors.affiliation: Laboratório de Imunopatologia, Instituto Butantan, Av. Dr. Vital Brasil 1500, 05503-900 São Paulo, SP, Brazil.
authors.forename: Daiana S
authors.initials: DS
authors.lastname: Lopes
authors.forename: Eliana
authors.initials: E
authors.lastname: Faquim-Mauro
authors.forename: Geraldo S
authors.initials: GS
authors.lastname: Magalhães
authors.forename: Iara C
authors.initials: IC
authors.lastname: Lima
authors.forename: Cristiani
authors.initials: C
authors.lastname: Baldo
authors.forename: Jay W
authors.initials: JW
authors.lastname: Fox
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Moura-da-Silva
authors.forename: Patricia B
authors.initials: PB
authors.lastname: Clissa
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000235:genetics / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D017837:Bothrops; D002460:Cell Line; D002470:Cell Survival; D003435:Crotalid Venoms / Q000633:toxicity*; D019040:E-Selectin / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects*; D006801:Humans; D007249:Inflammation / Q000139:chemically induced / Q000188:drug therapy; D018836:Inflammation Mediators / Q000494:pharmacology*; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism; D037181:Lectins, C-Type / Q000235:genetics / Q000378:metabolism; D053506:Matrix Metalloproteinase 10 / Q000235:genetics / Q000378:metabolism; D008666:Metalloendopeptidases / Q000633:toxicity*; D046228:Microarray Analysis / Q000379:methods; D009569:Nitric Oxide / Q000378:metabolism; D015854:Up-Regulation; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D000097146:Bothrops jararaca Venom; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D003435:Crotalid Venoms; D019040:E-Selectin; D018836:Inflammation Mediators; D016209:Interleukin-8; D037181:Lectins, C-Type; D053506:Matrix Metalloproteinase 10; D008666:Metalloendopeptidases; D009569:Nitric Oxide; D019010:Vascular Cell Adhesion Molecule-1; D000097146:Bothrops jararaca Venom; D000099192:CD69 Antigens; C511770:MMP10 protein, human