The effect of jararhagin, a metalloproteinase from Bothrops jararaca venom, on pro-inflammatory cytokines released by murine peritoneal adherent cells

The release of pro-inflammatory cytokines (IL-1 beta, IL-6 and TNF-alpha) f rom murine peritoneal adherent cells (MPAC) was studied after exposure to j ararhagin, a metalloproteinase/disintegrin isolated from Bothrops jararaca venom. MPACs were treated with LPS (lipopolysaccharide), jararhagin, or EDT A-inactivated jararhagin for up to 24 h. Following incubation, the culture supernatant was assayed by ELISA for the presence of cytokines, while the c ells were analysed for viability and cytokine mRNA expression. The cells ex posed to native jararhagin released TNF-alpha and IL-1 beta after 4 and 24 h respectively. When MPACs were exposed to Jararhagin treated with EDTA, TN F-alpha and IL-1 beta production was sustained throughout the culture perio d and IL-6 production was observed. TNF-alpha, IL-6 and IL-1 beta mRNA were detected 4 h after stimulation with either native or EDTA-treatedjararhagi n. Addition of jararhagin to LPS stimulated cells resulted in a dramatic de crease in the release of IL-6 and TNF-alpha. RT-PCR showed that this inhibi tion does not occur at the transcriptional level and further experiments sh owed that jararhagin degraded soluble cytokines by proteolytic activity. Th is study suggests that jararhagin induces TNF-alpha, IL-1 beta and IL-6 exp ression, which may be rapidly degraded by its proteolytic activity. (C) 200 1 Elsevier Science Ltd. All rights reserved.