Effect of endocannabinoids on IgE-mediated allergic response in RBL-2H3 cells

Abstract

Recently, some endocannabinoids were reported to show anti-inflammatory and anti-allergic activities. In this respect, various arachidonoyl endocannabinoids were screened for the inhibition of allergic response in IgE-activated RBL-2H3 cells. Among arachidonoyl endocannabinoids with a low cytotoxicity, only NA-5HT remarkably inhibited the release of β-hexosaminidase (IC(50), 13.58 μM), a marker of degranulation, and tumor necrosis factor-α (IC(50), 12.52 μM), a pro-inflammatory cytokine, in IgE-activated RBL-2H3 cells. Additionally, NA-5HT markedly suppressed the formation of prostaglandin D(2) (PGD(2)) with IC(50) value of 1.27 μM and leukotriene B(4) (LTB(4)) with IC(50) value of 1.20 μM, and slightly LTC4. When effect of NA-5HT on early stage of FcεRI cascade was investigated, it significantly inhibited phosphorylation of Syk, but not Lyn. Furthermore, NA-5HT suppressed phosphorylation of PLCγ1/2 and PKCδ, related to degranulation process, as well as phosphorylation of LAT, ERK1/2, p38, JNK, Gab2, PI3K and Akt, implicated in the expression of pro-inflammatory cytokines. Relative to its effect on the late stage, NA-5HT slightly reduced phosphorylation of 5-lipoxygenase (5-LO) and cyclooxygenase-2 (COX-2). Additionally, NA-5HT significantly reduced the level of p40(phox), and partially inhibited the expression of p47(phox) and p67(phox). From these results, it is suggested that NA-5HT expresses anti-allergic action by suppressing the activation of Syk, LAT, p38, JNK, PI3K and Akt, as well as the expression of ERK1/2 and NADPH oxidase subunits. Further, a strong inhibition of PGD(2) or LTB(4) biosynthesis by NA-5HT may be an additional mechanism for its anti-allergic action. Such anti-allergic actions of NA-5HT may contribute to further information about its biological functions.

Keywords: 2-AG; 2-arachidonoyl glycerol; 5-LO; 5-lipoxygenase; Akt; Allergic response; CB; COX-2; ERK1/2; FAAH; GRB2-associated binding protein 2; Gab2; HSA; IgE; Immunoglobulin E; JNK; LAT; LTB(4); LTC(4); Lyn; N-arachidonoyl glycine; N-arachidonoyl serine; N-arachidonoyl serotonin; N-arachidonoyl taurine; N-arachidonoyl γ-aminobutyric acid; NA-5HT; NA-GABA; NA-Gly; NA-Ser; NA-Tau; PGD(2); PI3K; PKCδ; PLCγ; Phosphatidylinositol 3-kinase; RBL-2H3 cells; Src family protein kinase; Syk; TNF; TRPV; c-Jun N-terminal kinase; cPLA(2); cannabinoid; cyclooxygenase-2; cytosolic phospholipase A(2); extracellular signal-regulated kinase 1/2; fatty acid amide hydrolase; human serum albumin; leukotriene B(4); leukotriene C(4); linker for activation of T cells; phospholipase Cγ; prostaglandin D(2); protein kinase B; protein kinase Cδ; spleen tyrosine kinase; transient receptor potential vanilloid; tumor necrosis factor.