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) from murine peritoneal adherent cells (MPAC) was studied after exposure to jararhagin, a metalloproteinase/disintegrin isolated from Bothrops jararaca venom. MPACs were treated with LPS (lipopolysaccharide), jararhagin, or EDTA-inactivated jararhagin for up to 24 h. Following incubation, the culture supernatant was assayed by ELISA for the presence of cytokines, while the cells were analysed for viability and cytokine mRNA expression. The cells exposed 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, TNF-alpha and IL-1 beta production was sustained throughout the culture period 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-treatedjararhagin. Addition of jararhagin to LPS stimulated cells resulted in a dramatic decrease in the release of IL-6 and TNF-alpha. RT-PCR showed that this inhibition does not occur at the transcriptional level and further experiments showed that jararhagin degraded soluble cytokines by proteolytic activity. This study suggests that jararhagin induces TNF-alpha, IL-1 beta and IL-6 expression, which may be rapidly degraded by its proteolytic activity. (C) 2001 Elsevier Science Ltd. All rights reserved.