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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) on degranulation, cytokine levels, and [Ca2+]i elevation of the mBMMCs that were stimulated with DNP-BSA. (a) The time scheme for the degranulation assay, real-time PCR analysis, and transcriptome analysis. (b) The mBMMCs that were sensitized with 1.5 μg/mL anti-DNP IgE (6 h) were incubated with ZF for 30 min. The cells were stimulated with (filled) or without (open) DNP-BSA (1 h), and the β-hexosaminidase release was determined. (c) The mBMMCs were pretreated with ZF (0.32 mg/mL) for 90 min and stained with PI; the viability was analyzed using the FACS Calibur system. (d) The sensitized mBMMCs were incubated with ZF (0.32 mg/mL) or vehicle for 30 min and then stimulated with DNP-BSA for 1 h, and the total RNA was extracted. The mRNA levels of TNF-α, IL-4, and IL-13 were analyzed by real-time PCR. The results are expressed as the relative ratio to the vehicle-treated mBMMCs without DNP-BSA stimulation. The data are expressed as the mean ± SD (n = 3–4; b, c, d). #P < 0.05 compared with DNP-BSA (−) (d), *P < 0.05 compared with the vehicle (b, d). (e) The sensitized mBMMCs were labeled with Fura-2 AM for 30 min and incubated with ZF or vehicle for 30 min. Ca2+-mobilization was determined following stimulation with DNP-BSA. The data are representative of at least three independent experiments.
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fig2: The effect of ZF (0.32 mg/mL) on degranulation, cytokine levels, and [Ca2+]i elevation of the mBMMCs that were stimulated with DNP-BSA. (a) The time scheme for the degranulation assay, real-time PCR analysis, and transcriptome analysis. (b) The mBMMCs that were sensitized with 1.5 μg/mL anti-DNP IgE (6 h) were incubated with ZF for 30 min. The cells were stimulated with (filled) or without (open) DNP-BSA (1 h), and the β-hexosaminidase release was determined. (c) The mBMMCs were pretreated with ZF (0.32 mg/mL) for 90 min and stained with PI; the viability was analyzed using the FACS Calibur system. (d) The sensitized mBMMCs were incubated with ZF (0.32 mg/mL) or vehicle for 30 min and then stimulated with DNP-BSA for 1 h, and the total RNA was extracted. The mRNA levels of TNF-α, IL-4, and IL-13 were analyzed by real-time PCR. The results are expressed as the relative ratio to the vehicle-treated mBMMCs without DNP-BSA stimulation. The data are expressed as the mean ± SD (n = 3–4; b, c, d). #P < 0.05 compared with DNP-BSA (−) (d), *P < 0.05 compared with the vehicle (b, d). (e) The sensitized mBMMCs were labeled with Fura-2 AM for 30 min and incubated with ZF or vehicle for 30 min. Ca2+-mobilization was determined following stimulation with DNP-BSA. The data are representative of at least three independent experiments.

Mentions: In a dose-dependent manner, ZF significantly inhibited the degranulation of the mBMMCs that were induced with DNP-BSA (Figure 2(b)) without detectably affecting the viability of the mBMMCs (Figure 2(c)). As shown in Figure 2(d), mRNA expression of TNF-α, IL-4, and IL-13 was extremely enhanced by DNP-BSA. ZF (0.32 mg/mL) significantly suppressed the increased expression of TNF-α, IL-4, and IL-13. In contrast, the expression of these genes was not affected by ZF in the mBMMCs that were not stimulated with DNP-BSA (unactivated mBMMCs). In addition, ZF attenuated the DNP-BSA-induced [Ca2+]i increases (Figure 2(e)).


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) on degranulation, cytokine levels, and [Ca2+]i elevation of the mBMMCs that were stimulated with DNP-BSA. (a) The time scheme for the degranulation assay, real-time PCR analysis, and transcriptome analysis. (b) The mBMMCs that were sensitized with 1.5 μg/mL anti-DNP IgE (6 h) were incubated with ZF for 30 min. The cells were stimulated with (filled) or without (open) DNP-BSA (1 h), and the β-hexosaminidase release was determined. (c) The mBMMCs were pretreated with ZF (0.32 mg/mL) for 90 min and stained with PI; the viability was analyzed using the FACS Calibur system. (d) The sensitized mBMMCs were incubated with ZF (0.32 mg/mL) or vehicle for 30 min and then stimulated with DNP-BSA for 1 h, and the total RNA was extracted. The mRNA levels of TNF-α, IL-4, and IL-13 were analyzed by real-time PCR. The results are expressed as the relative ratio to the vehicle-treated mBMMCs without DNP-BSA stimulation. The data are expressed as the mean ± SD (n = 3–4; b, c, d). #P < 0.05 compared with DNP-BSA (−) (d), *P < 0.05 compared with the vehicle (b, d). (e) The sensitized mBMMCs were labeled with Fura-2 AM for 30 min and incubated with ZF or vehicle for 30 min. Ca2+-mobilization was determined following stimulation with DNP-BSA. The data are representative of at least three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3375181&req=5

fig2: The effect of ZF (0.32 mg/mL) on degranulation, cytokine levels, and [Ca2+]i elevation of the mBMMCs that were stimulated with DNP-BSA. (a) The time scheme for the degranulation assay, real-time PCR analysis, and transcriptome analysis. (b) The mBMMCs that were sensitized with 1.5 μg/mL anti-DNP IgE (6 h) were incubated with ZF for 30 min. The cells were stimulated with (filled) or without (open) DNP-BSA (1 h), and the β-hexosaminidase release was determined. (c) The mBMMCs were pretreated with ZF (0.32 mg/mL) for 90 min and stained with PI; the viability was analyzed using the FACS Calibur system. (d) The sensitized mBMMCs were incubated with ZF (0.32 mg/mL) or vehicle for 30 min and then stimulated with DNP-BSA for 1 h, and the total RNA was extracted. The mRNA levels of TNF-α, IL-4, and IL-13 were analyzed by real-time PCR. The results are expressed as the relative ratio to the vehicle-treated mBMMCs without DNP-BSA stimulation. The data are expressed as the mean ± SD (n = 3–4; b, c, d). #P < 0.05 compared with DNP-BSA (−) (d), *P < 0.05 compared with the vehicle (b, d). (e) The sensitized mBMMCs were labeled with Fura-2 AM for 30 min and incubated with ZF or vehicle for 30 min. Ca2+-mobilization was determined following stimulation with DNP-BSA. The data are representative of at least three independent experiments.
Mentions: In a dose-dependent manner, ZF significantly inhibited the degranulation of the mBMMCs that were induced with DNP-BSA (Figure 2(b)) without detectably affecting the viability of the mBMMCs (Figure 2(c)). As shown in Figure 2(d), mRNA expression of TNF-α, IL-4, and IL-13 was extremely enhanced by DNP-BSA. ZF (0.32 mg/mL) significantly suppressed the increased expression of TNF-α, IL-4, and IL-13. In contrast, the expression of these genes was not affected by ZF in the mBMMCs that were not stimulated with DNP-BSA (unactivated mBMMCs). In addition, ZF attenuated the DNP-BSA-induced [Ca2+]i increases (Figure 2(e)).

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