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Sphingosine kinase-1-dependent and -independent inhibitory effects of zanthoxyli fructus to attenuate the activation of mucosal mast cells and ameliorate food allergies in mice.

Wang X, Kageyama-Yahara N, Hayashi S, Yamamoto T, Kadowaki M - Evid Based Complement Alternat Med (2012)

Bottom Line: As a result, both antigen-induced and calcium ionophore-induced degranulation was significantly inhibited by Zanthoxyli Fructus water extract (ZF) in mucosal-type murine bone marrow-derived mast cells (mBMMCs).Furthermore, ZF inhibited allergic symptoms in an ovalbumin-caused murine FA model and decreased the number of infiltrating mucosal mast cells and the enhanced mRNA expression levels of IL-4 and Sphk1 in the FA mice colons.These results indicate that ZF suppresses mucosal mast cell activities mainly through Sphk1-dependent mechanism, and ZF is utilized for the development of a novel, potent anti-FA agent.

View Article: PubMed Central - PubMed

Affiliation: Division of Gastrointestinal Pathophysiology, Institute of Natural Medicine, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan.

ABSTRACT
Food allergy (FA) is relatively a common disease in infants, but effective drug therapies are not yet available. Notably, mucosal mast cells, but not connective-tissue mast cells, play important roles in food allergic reactions via the release of inflammatory mediators. Therefore, we screened medicinal herb extracts for in vitro and in vivo antiallergic activity through inhibiting mucosal mast cell activation. As a result, both antigen-induced and calcium ionophore-induced degranulation was significantly inhibited by Zanthoxyli Fructus water extract (ZF) in mucosal-type murine bone marrow-derived mast cells (mBMMCs). ZF suppressed the antigen-induced [Ca(2+)](i) elevation and the antigen-enhanced mRNA expression of TNF-α, IL-4, and IL-13. The transcriptome and real-time PCR analyses revealed that ZF greatly decreased the antigen-enhanced expression level of sphingosine kinase 1 (Sphk1), which plays a key role in the FcεRI-mediated immune responses in mast cells. Furthermore, ZF inhibited allergic symptoms in an ovalbumin-caused murine FA model and decreased the number of infiltrating mucosal mast cells and the enhanced mRNA expression levels of IL-4 and Sphk1 in the FA mice colons. These results indicate that ZF suppresses mucosal mast cell activities mainly through Sphk1-dependent mechanism, and ZF is utilized for the development of a novel, potent anti-FA agent.

No MeSH data available.


Related in: MedlinePlus

The effect of ZF (0.32 mg/mL) treatment on A23187-induced [Ca2+]i elevation and degranulation. (a) The time scheme for the degranulation assay. (b) RBL-2H3 cells were labeled with Fluo-3 AM and incubated with ZF or vehicle for 30 min. The cells were stimulated with 25 μM A23187 and monitored by calcium imaging. The data are representative of at least three independent experiments. (c) The RBL-2H3 cells were incubated with ZF or vehicle for 30 min, the cells were stimulated with (filled) or without (open) A23187 for 30 min, and β-hexosaminidase release was determined. (d) The mBMMCs were incubated with ZF (0.32 mg/mL), DMS (10 μM), cyclosporin A (1 μM), W-7 (32 μM), KN-93 (32 μM), or vehicle for 30 min, and the cells were stimulated with 25 μM A23187 for 30 min. β-hexosaminidase release was determined. The data are expressed as the mean ± SD (n = 4; c, d). *P < 0.05 compared with the vehicle (c, d).
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fig5: The effect of ZF (0.32 mg/mL) treatment on A23187-induced [Ca2+]i elevation and degranulation. (a) The time scheme for the degranulation assay. (b) RBL-2H3 cells were labeled with Fluo-3 AM and incubated with ZF or vehicle for 30 min. The cells were stimulated with 25 μM A23187 and monitored by calcium imaging. The data are representative of at least three independent experiments. (c) The RBL-2H3 cells were incubated with ZF or vehicle for 30 min, the cells were stimulated with (filled) or without (open) A23187 for 30 min, and β-hexosaminidase release was determined. (d) The mBMMCs were incubated with ZF (0.32 mg/mL), DMS (10 μM), cyclosporin A (1 μM), W-7 (32 μM), KN-93 (32 μM), or vehicle for 30 min, and the cells were stimulated with 25 μM A23187 for 30 min. β-hexosaminidase release was determined. The data are expressed as the mean ± SD (n = 4; c, d). *P < 0.05 compared with the vehicle (c, d).

Mentions: To investigate the pharmacological profile of ZF on mucosal mast cell activation, we examined the effect of ZF on the [Ca2+]i increases and β-hexosaminidase release induced by the calcium ionophore A23187. The ZF treatment diminished the [Ca2+]i increases in RBL-2H3 cells (Figure 5(b)). In addition, ZF significantly inhibited A23187-induced degranulation in both RBL-2H3 cells and mBMMCs (Figures 5(c), 5(d)). Moreover, the calcineurin inhibitor cyclosporin A (1 μM), the calmodulin antagonist W-7 (32 μM), and the Ca2+/calmodulin-dependent protein kinase II inhibitor KN-93 (32 μM) inhibited A23187-induced degranulation in mBMMCs to a similar extent as ZF, which indicates that A23187 induces degranulation via both calcineurin- and calmodulin-dependent pathways in mBMMCs (Figure 5(d)). In contrast, DMS (10 μM) did not affect A23187-induced degranulation in mBMMCs (Figure 5(d)). Furthermore, when the mBMMCs were stimulated with the calcium ionophore A23187 (instead of anti-DNP IgE and DNP-BSA) and examined with the Affymetrix Mouse Genechip Array, the expression of Sphk1 was not altered (data not shown).


Sphingosine kinase-1-dependent and -independent inhibitory effects of zanthoxyli fructus to attenuate the activation of mucosal mast cells and ameliorate food allergies in mice.

Wang X, Kageyama-Yahara N, Hayashi S, Yamamoto T, Kadowaki M - Evid Based Complement Alternat Med (2012)

The effect of ZF (0.32 mg/mL) treatment on A23187-induced [Ca2+]i elevation and degranulation. (a) The time scheme for the degranulation assay. (b) RBL-2H3 cells were labeled with Fluo-3 AM and incubated with ZF or vehicle for 30 min. The cells were stimulated with 25 μM A23187 and monitored by calcium imaging. The data are representative of at least three independent experiments. (c) The RBL-2H3 cells were incubated with ZF or vehicle for 30 min, the cells were stimulated with (filled) or without (open) A23187 for 30 min, and β-hexosaminidase release was determined. (d) The mBMMCs were incubated with ZF (0.32 mg/mL), DMS (10 μM), cyclosporin A (1 μM), W-7 (32 μM), KN-93 (32 μM), or vehicle for 30 min, and the cells were stimulated with 25 μM A23187 for 30 min. β-hexosaminidase release was determined. The data are expressed as the mean ± SD (n = 4; c, d). *P < 0.05 compared with the vehicle (c, d).
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Related In: Results  -  Collection

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fig5: The effect of ZF (0.32 mg/mL) treatment on A23187-induced [Ca2+]i elevation and degranulation. (a) The time scheme for the degranulation assay. (b) RBL-2H3 cells were labeled with Fluo-3 AM and incubated with ZF or vehicle for 30 min. The cells were stimulated with 25 μM A23187 and monitored by calcium imaging. The data are representative of at least three independent experiments. (c) The RBL-2H3 cells were incubated with ZF or vehicle for 30 min, the cells were stimulated with (filled) or without (open) A23187 for 30 min, and β-hexosaminidase release was determined. (d) The mBMMCs were incubated with ZF (0.32 mg/mL), DMS (10 μM), cyclosporin A (1 μM), W-7 (32 μM), KN-93 (32 μM), or vehicle for 30 min, and the cells were stimulated with 25 μM A23187 for 30 min. β-hexosaminidase release was determined. The data are expressed as the mean ± SD (n = 4; c, d). *P < 0.05 compared with the vehicle (c, d).
Mentions: To investigate the pharmacological profile of ZF on mucosal mast cell activation, we examined the effect of ZF on the [Ca2+]i increases and β-hexosaminidase release induced by the calcium ionophore A23187. The ZF treatment diminished the [Ca2+]i increases in RBL-2H3 cells (Figure 5(b)). In addition, ZF significantly inhibited A23187-induced degranulation in both RBL-2H3 cells and mBMMCs (Figures 5(c), 5(d)). Moreover, the calcineurin inhibitor cyclosporin A (1 μM), the calmodulin antagonist W-7 (32 μM), and the Ca2+/calmodulin-dependent protein kinase II inhibitor KN-93 (32 μM) inhibited A23187-induced degranulation in mBMMCs to a similar extent as ZF, which indicates that A23187 induces degranulation via both calcineurin- and calmodulin-dependent pathways in mBMMCs (Figure 5(d)). In contrast, DMS (10 μM) did not affect A23187-induced degranulation in mBMMCs (Figure 5(d)). Furthermore, when the mBMMCs were stimulated with the calcium ionophore A23187 (instead of anti-DNP IgE and DNP-BSA) and examined with the Affymetrix Mouse Genechip Array, the expression of Sphk1 was not altered (data not shown).

Bottom Line: As a result, both antigen-induced and calcium ionophore-induced degranulation was significantly inhibited by Zanthoxyli Fructus water extract (ZF) in mucosal-type murine bone marrow-derived mast cells (mBMMCs).Furthermore, ZF inhibited allergic symptoms in an ovalbumin-caused murine FA model and decreased the number of infiltrating mucosal mast cells and the enhanced mRNA expression levels of IL-4 and Sphk1 in the FA mice colons.These results indicate that ZF suppresses mucosal mast cell activities mainly through Sphk1-dependent mechanism, and ZF is utilized for the development of a novel, potent anti-FA agent.

View Article: PubMed Central - PubMed

Affiliation: Division of Gastrointestinal Pathophysiology, Institute of Natural Medicine, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan.

ABSTRACT
Food allergy (FA) is relatively a common disease in infants, but effective drug therapies are not yet available. Notably, mucosal mast cells, but not connective-tissue mast cells, play important roles in food allergic reactions via the release of inflammatory mediators. Therefore, we screened medicinal herb extracts for in vitro and in vivo antiallergic activity through inhibiting mucosal mast cell activation. As a result, both antigen-induced and calcium ionophore-induced degranulation was significantly inhibited by Zanthoxyli Fructus water extract (ZF) in mucosal-type murine bone marrow-derived mast cells (mBMMCs). ZF suppressed the antigen-induced [Ca(2+)](i) elevation and the antigen-enhanced mRNA expression of TNF-α, IL-4, and IL-13. The transcriptome and real-time PCR analyses revealed that ZF greatly decreased the antigen-enhanced expression level of sphingosine kinase 1 (Sphk1), which plays a key role in the FcεRI-mediated immune responses in mast cells. Furthermore, ZF inhibited allergic symptoms in an ovalbumin-caused murine FA model and decreased the number of infiltrating mucosal mast cells and the enhanced mRNA expression levels of IL-4 and Sphk1 in the FA mice colons. These results indicate that ZF suppresses mucosal mast cell activities mainly through Sphk1-dependent mechanism, and ZF is utilized for the development of a novel, potent anti-FA agent.

No MeSH data available.


Related in: MedlinePlus