The
snake venom MT-III is a group IIA
secreted phospholipase A2 (
sPLA2)
enzyme with functional and structural similarities with mammalian pro-inflammatory
sPLA2s of the same group. Previously, we demonstrated that
MT-III directly activates the
innate inflammatory response of macrophages, including release of inflammatory mediators and formation of lipid droplets (LDs). However, the mechanisms coordinating these processes remain unclear. In the present study, by using TLR2-/- or MyD88-/- or C57BL/6 (WT) male mice, we report that TLR2 and MyD88 signaling have a critical role in
MT-III-induced inflammatory response in macrophages.
MT-III caused a marked release of
PGE2,
PGD2,
PGJ2, IL-1β and
IL-10 and increased the number of LDs in WT macrophages. In
MT-III-stimulated TLR2-/- macrophages, formation of LDs and release of
eicosanoids and
cytokines were abrogated. In MyD88-/- macrophages,
MT-III-induced release of
PGE2, IL-1β and
IL-10 was abrogated, but release of
PGD2 and
PGJ2 was maintained. In addition, COX-2
protein expression seen in
MT-III-stimulated WT macrophages was abolished in both TLR2-/- and MyD88-/- cells, while
perilipin 2 expression was abolished only in MyD88-/- cells. We further demonstrated a reduction of saturated, monounsaturated and
polyunsaturated fatty acids and a release of the TLR2 agonists palmitic and
oleic acid from
MT-III-stimulated WT macrophages compared with WT control cells, thus suggesting these
fatty acids as major messengers for
MT-III-induced engagement of TLR2/MyD88 signaling. Collectively, our findings identify for the first time a TLR2 and MyD88-dependent mechanism that underlies group IIA sPLA2-induced inflammatory response in macrophages.