Metalloproteinase Inhibitors Decrease The Loss of Human Keratinocytes Cell Viability Induced By Sphingomyelinases from Loxosceles intermedia Spider Venom

¹Paixão-Cavalcante, D.; ¹Gonçalvez-de-Andrade, R.M.; ¹Fernandes Pedrosa, M.F.; ²van den Berg C.W.  and ¹Tambourgi, D.V.

1 Lab. Imunoquímica, Instituto Butantan, São Paulo, Brazil; 2 Dep. of Pharmacology, Therapeutics and Toxicology, University of Wales, Cardiff, UK.

The Loxosceles spider genus is the main responsible for arachnid accidents in Brazil. The victims of these spiders usually show an ulcer in the bite site of difficult healing. Our previous studies have shown the participation of endogenous metalloproteinases in the dermonecrotic lesion development. The specific treatment for the dermonecrosis is still not avaiable. The aim of this study was to evaluate the ability of tetracycline derived metalloproteinases inhibitors in preventing the loss of human keratinocytes cell viability induced by in vitro treatment of the cells with Loxosceles intermedia (Li) venom or recombinant sphingomyelinase P2 (recP2). Human keratinocytes, HaCaT cell line, were cultivated in 24 well plates (10⁵cell/mL). Before starting the treatment, cells were cultivated without fetal bovine serum and then incubated with 20mg/mL of Li venom or recP2 in the presence or not of increasing concentrations of tetracyclin, doxycycline and minocycline, during three days. On the third day the cell viability was analyzed by Alamar Blue assay. The tetracycline derived inhibitors were able to reduce the loss of keratinocytes cell viability, provoked by Li venom or recP2, at 30, 40 and 50 mg/mL concentrations, being the most efficient effect obtained by tetracyclin treatment. Therefore, tetracyclin derived metalloproteinases inhibitors are good candidates for controlling dermonecrotic lesions induced by Loxosceles envenomation.

Support: FAPESP, the Wellcome Trust, CNPq

Correspondence to: danipaixao@butantan.gov.br