A new method to evaluate anti-allergic effect of food component by measuring leukotriene B4 from a mouse mast cell line

Abstract

Leukotrienes (LTs), chemical mediators produced by mast cells, play an important role in allergic symptoms such as food allergies and hay fever. We tried to construct an evaluation method for the anti-LTB activity of chemical substances using a mast cell line, PB-3c. PB-3c pre-cultured with or without arachidonic acid (AA) was stimulated by calcium ionophore (A23187) for 20 min, and LTB production by the cells was determined by HPLC with UV detection. LTB was not detected when PB-3c was pre-cultured without AA. On the other hand, LTB production by PB-3c pre-cultured with AA was detectable by HPLC, and the optimal conditions of PB-3c for LTB detection were to utilize the cells pre-cultured with 50 µM AA for 48 h. MK-886 (5-lipoxygenase inhibitor) completely inhibited LTB production, but AACOCF (phospholipase A inhibitor) slightly increased LTB production, suggesting that LTB was generated from exogenous free AA through 5-lipoxygenase pathway. We applied this technique to the evaluation of the anti-LTB activity of food components. PB-3c pre-cultured with 50 µM AA for 48 h was stimulated with A23187 in the presence of 50 µM soybean isoflavones (daidzin, genistin, daidzein, and genistein), equol, quercetin, or kaempferol. Genistein, equol, quercetin, and kaempferol strongly inhibited LTB production without cytotoxicity. These results suggest that a new assay system using PB-3c is convenient to evaluate LTB inhibition activity by food components. This method could be utilized for elucidation of the mechanisms of LTB release suppression by food components such as flavonoids and the structure-activity relationship.

Long Chain Polyunsaturated Fatty Acids (LCPUFAs) in the Prevention of Food Allergy

“…Quercetin can be found in red wine, apples, green tea and onion and contains both anti-inflammatory and anti-oxidative properties (69). It has been reported to inhibit leukotriene B4 levels in MCs (70), reduce the gene expression of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-8) (71) and suppress inflammation in IgE-mediated intestinal epithelial cell (Caco-2) and rat basophil (RBL-2H3) activation models (72). Flavonoids have also been shown to inhibit allergic effector cells such as MCs known to contribute to allergic symptoms (73).…”

Protective Effect of Genistein against Compound 48/80 Induced Anaphylactoid Shock via Inhibiting MAS Related G Protein-Coupled Receptor X2 (MRGPRX2)

“…Genistein is a non-steroidal polyphenol that possesses diverse pharmacological activities. There is evidence for the protective effects of Genistein in inflammation [21][22][23], diabetes [24,25], cancer [26,27] and hypersensitivity reactions [28,29]. In a recent paper, Kim, Dong Hwan et al reported the anti-inflammatory activity of Genistein on mast cells via inhibiting cytokines and the ERK pathway and proposed potential of Genistein for the treatment of allergic inflammation and anaphylactic shock [30].…”

Targeting Mammalian 5-Lipoxygenase by Dietary Phenolics as an Anti-Inflammatory Mechanism: A Systematic Review

“…Curcumin also targeted LTC 4 biosynthesis in mast cells by blocking 5-LOX translocation and cPLA 2 activation [ 102 ]. Quercetin reduced biosynthesis of LTB 4 in RBL-1 basophils, neutrophils, and murine PB-3c mast cells [ 146 , 147 , 148 ], while genistein acted on eosinophils decreasing LTC 4 synthesis via inhibition of 5-LOX translocation [ 95 ]. Likewise, the effect of silibinin on macrophages (Kupffer cells) was associated with lower LTB 4 levels while sparing the COX-2 pathway [ 149 , 150 ].…”