Korean J Physiol Pharmacol.  2021 May;25(3):251-258. 10.4196/kjpp.2021.25.3.251.

Flos magnoliae constituent fargesin has an anti-allergic effect via ORAI1 channel inhibition

• Hong PTL
• Kim HJ
• Kim WK
• Nam JH

• KMID: 2514950
• DOI: http://doi.org/10.4196/kjpp.2021.25.3.251

Abstract

Flos magnoliae (FM), the dry flower buds of Magnolia officinalis or its related species, is a traditional herbal medicine commonly used in Asia for symptomatic relief of and treating allergic rhinitis, headache, and sinusitis. Although several studies have reported the effects of FM on store-operated calcium entry (SOCE) via the ORAI1 channel, which is essential during intracellular calcium signaling cascade generation for T cell activation and mast cell degranulation, the effects of its isolated constituents on SOCE remain unidentified. Therefore, we investigated which of the five major constituents of 30% ethanoic FM (vanillic acid, tiliroside, eudesmin, magnolin, and fargesin) inhibit SOCE and their physiological effects on immune cells. The conventional whole-cell patch clamp results showed that fargesin, magnolin, and eudesmin significantly inhibited SOCE and thus human primary CD4 ⁺ T lymphocyte proliferation, as well as allergen-induced histamine release in mast cells. Among them, fargesin demonstrated the most potent inhibitory effects not only on ORAI1 (IC ₅₀ = 12.46 ± 1.300 μM) but also on T-cell proliferation (by 87.74% ± 1.835%) and mast cell degranulation (by 20.11% ± 5.366%) at 100 μM. Our findings suggest that fargesin can be a promising candidate for the development of therapeutic drugs to treat allergic diseases.

Keyword

Eudesmin; Fargesin; Mast cell; ORAI; T Lymphocyte

Figure

• 

  Fig. 1 Inhibitory effects of five major constituent chemical compounds in 30% Flos magnoliae ethanolic extract (FMEtOH) on the ORAI1 current (IORAI1) in HEK293T cells co-expressing ORAI1 and stromal interaction molecule 1 (STIM1). (A) Representative current (I)–voltage (V) relationship curve showing IORAI1 by various constituents (100 µM) and BTP2. (B) Normalized IORAI1 inhibition histograms indicating treatment with various constituents (100 µM) at −120 mV. Values are presented as mean ± SEM. *p < 0.05 and ****p < 0.0001 compared to control (IORAI1).

• 

  Fig. 2 Effects of fargesin on ORAI1 current (IORAI1). (A) Representative trace chart depicting dose-dependent fargesin inhibition of IORAI1 at 1, 3, 10, 30, 100, and 300 µM. (B) Typical current–voltage (I–V) relationships describing inhibitory proportion of fargesin at a range of concentrations (1–300 µM). (C) Concentration dependence of IORAI1 inhibition by fargesin at −120 mV and fitted dose-response curves. Values are presented as mean ± SEM.

• 

  Fig. 3 Inhibitory effects of fargesin and eudesmin on β-hexosaminidase activity released from RBL-2H3 cells. (A) β-Hexosaminidase activity in the media after the exposure of IgE-antigen (Ag)-stimulated mast cells to 3, 10, 30, and 100 µM fargesin and 10 µM BTP2. (B) The release of β-hexosaminidase in stimulated cells after exposure to 3, 10, 30, and 100 µM eudesmin and 10 µM BTP2. Values are presented as means ± SEM. *p < 0.05, ****p < 0.0001 vs. control (value of β-hexosaminidase release in stimulated cells without administration of constituent compounds).

• 

  Fig. 4 Inhibitory effects of fargesin, eudesmin, and magnolin on CD3/CD28 co-stimulated human CD4+ T lymphocyte activation. (A) Fargesin toxicity to Jurkat T cell line after administration of 1, 3, 10, 30, and 100 μM based on CCK-8 assay. (B) Eudesmin cytotoxicity to Jurkat T cell at concentrations in (A). (C) Cytotoxic effects of magnolin to Jurkat T cell at concentrations in (A). (D) Representative figure illustrating proliferation of human naïve CD4+ T cells treated with three constituents, and percentage T cell growth inhibition determined by carboxyfluorescein succinimidyl ester (CFSE). Human naïve CD4+ T cells stimulated with antibodies CD3 and CD28 (negative control); CD3/CD28-stimulated cells treated with 10 μM BTP2 (positive control). (E) Statistical analysis of percentage CD4+ T cell proliferation with constituent and BTP2 treatments in cells stimulated or not stimulated by CD3/CD28. ****p < 0.0001 vs. the control.

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