Salicylaldehyde Suppresses IgE-Mediated Activation of Mast Cells and Ameliorates Anaphylaxis in Mice

Abstract

Mast cells (MCs) play key roles in IgE-mediated immunoresponses, including in the protection against parasitic infections and the onset and/or symptoms of allergic diseases. IgE-mediated activation induces MCs to release mediators, including histamine and leukotriene, as an early response, and to produce cytokines as a late phase response. Attempts have been made to identify novel antiallergic compounds from natural materials such as Chinese medicines and food ingredients. We herein screened approximately 60 compounds and identified salicylaldehyde, an aromatic aldehyde isolated from plant essential oils, as an inhibitor of the IgE-mediated activation of MCs. A degranulation assay, flow cytometric analyses, and enzyme-linked immunosorbent assays revealed that salicylaldehyde inhibited the IgE-mediated degranulation and cytokine expression of bone-marrow-derived MCs (BMMCs). The salicylaldehyde treatment reduced the surface expression level of FcεRI, the high affinity receptor for IgE, on BMMCs, and suppressed the IgE-induced phosphorylation of tyrosine residues in intercellular proteins, possibly Lyn, Syk, and Fyn, in BMMCs. We also examined the effects of salicylaldehyde in vivo using passive anaphylaxis mouse models and found that salicylaldehyde administration significantly enhanced the recovery of a reduced body temperature due to systemic anaphylaxis and markedly suppressed ear swelling, footpad swelling, and vascular permeability in cutaneous anaphylaxis.

Kaempferol Suppresses the Activation of Mast Cells by Modulating the Expression of FcεRI and SHIP1

“…A degranulation assay was performed as previously described [ 36 , 37 ]. Briefly, BMMCs sensitized with anti-TNP mouse IgE (clone IgE-3, BD Biosciences, San Jose, CA, USA) were stimulated with TNP-BSA (LSL-LG1117, Cosmo Bio, Tokyo, Japan) in Tyrode’s buffer, and the culture supernatant and cells were harvested to determine β-hexosaminidase activity.…”

“…Western blot analyses using anti-phospho-tyrosine (clone 4G10, Merck, Darmstadt, Germany), anti-NRF2 (clone D1Z9C, Cell Signaling Technology, Danvers, MA, USA), anti-SHIP1 (clone V-19, Santa Cruz Biotechnology), anti-PLCγ1 (clone D9H10, Cell Signaling Technology), anti-phospho-PLCγ1 (Tyr783) (polyclonal, Cell Signaling Technology), and anti-β-actin (clone AC-15, Sigma-Aldrich, St. Louis, MO, USA) were performed as previously described [ 37 ]. Quantification of SHIP1 expression was carried out with Image J (NIH, Bethesda, MD, USA).…”

“…Quantitative PCR was performed using a Step-One real-time PCR system (Applied Biosystems, Waltham, MA, USA) with THUNDERBIRD SYBR qPCR Mix (TOYOBO) and primers. The primers used for measuring the mRNA levels of Fcer1a , Ms4a2 (encoding FcεRIβ), and Fcer1g were described previously [ 37 ]. To measure the mRNA levels of Inpp5d (SHIP1), Inppl1 (SHIP2), Nr0b2 (SHP1), and Ptpra (PTPa), the following primers were used: Inpp5d forward; 5′-tgcagtcaatatggaacatcaag-3′, Inpp5d reverse; 5′-gagacgaattgagatgtgactcc-3′, Inppl1 forward; 5′-tcagggtcactcatatcaaggtt-3′, Inppl1 reverse; 5′-cgagtagtcttctcattccctga-3′, Nr0b2 forward; 5′-ccacctatcgatttgaaagactg-3′, Nr0b2 reverse; 5′-cgtttacccgagtagcgtagtaa-3′, Ptpra forward; 5′-cccaatactggccagaccaa-3′, Ptpra reverse; 5′-cctcgacagacacacggacat-3′.…”

Kaempferol suppresses the activation of mast cells by modulating the expression of FcεRI and SHIP1

“…The concentrations of IL-6, and TNF-α were measured using ELISA kits (ELISA MAX series, BioLegend), and of IL-13 was determined using the mouse IL-13 DuoSet ELISA kit (DY413, R&D systems). ( Western blot analyses using anti-phospho-tyrosine (clone 4G10, Millipore), anti-NRF2 (clone D1Z9C, Cell Signaling Technology), anti-SHIP1 (clone V-19, Santa Cruz Biotechnology), and anti-β-actin (clone AC-15, Sigma-Aldrich) were performed as previously described [31].…”

“…Western blot analyses using anti-phospho-tyrosine (clone 4G10, Millipore), anti-NRF2 (clone D1Z9C, Cell Signaling Technology), anti-SHIP1 (clone V-19, Santa Cruz Biotechnology), and anti-β-actin (clone AC-15, Sigma-Aldrich) were performed as previously described [31].…”