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
Figure 2
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 ( 3–4; b, c, d). # compared with DNP-BSA (−) (d), * 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.
