鹌鹑蛋通过抑制PAR2介导的MAPK和NF-κB激活对肥大细胞脱颗粒的抑制作用。

Inhibitory effects of quail egg on mast cells degranulation by suppressing PAR2-mediated MAPK and NF-kB activation.

作者信息

Lianto Priscilia, Ogutu Fredrick O, Zhang Yani, He Feng, Che Huilian

机构信息

Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, P.R. China.

College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, P.R. China; and.

出版信息

Food Nutr Res. 2018 Jul 20;62. doi: 10.29219/fnr.v62.1084. eCollection 2018.

DOI:10.29219/fnr.v62.1084

PMID:30083085

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6060182/

Abstract

BACKGROUND

Quail egg (QE) has been reported to possess an anti-allergic and anti-inflammatory activity. We have demonstrated that whole QE was able to attenuate the allergic symptoms in food allergy-induced EoE murine model, but whether QE albumen or QE yolk plays a more important role still remains unclear.

OBJECTIVE

In this current study, we investigated the suppressive role of QE in mast cell degranulation and cytokine production of the effect phase response.

METHOD

A passive cutaneous anaphylaxis (PCA) mouse model was used to confirm the anti-allergic effect of QE. Besides, HMC-1 cell model was used to study its suppressive role in more detail. In this study, we divided QE into three groups: whole QE, QE albumen, and QE yolk. The effect of QE treatment on mast cell degranulation and intracellular calcium influx was investigated. Moreover, the effect of QE allergy- related mediators, genes, and proteins were also assessed by ELISA, RT-PCR, and western blotting.

RESULTS AND DISCUSSION

Our data showed that the extent of mast cell degranulation-mediated ear vascular permeability in IgE-mediated PCA mice treated with whole QE (17 mg/kg) was decreased significantly up to 43.31 ± 0.42% reduction. HMC-1 cell-based immunological assay in vitro indicated that QE, particularly its albumen, acted as a 'mast cell stabilizer'. Under the concentration of 70 μg/mL, QE albumen effectively suppressed the releases of β-hexosaminidase, histamine, and tryptase, as well as Th2 and pro-inflammatory cytokine production; reached 30 up to 50% reduction. Besides, QE albumen was also able to significantly modulate the upregulation of IL-10 up to 58.30 ± 5.9%. Interestingly, our data indicated that QE yolk still had a significant inhibitory effect on modulating Th2 cytokines in its highest concentration (100 μg/mL), while QE albumen showed no inhibitory effect. Western blot analysis showed QE albumen effectively down-regulated the expressions of calcium-related protein (TRPC1, Orai1, STIM1, PLC-γ and IP3R), facilitated the reduction of PAR-2 and induced the reduction of phosphorylation of JNK, IKKα, p50 and p65 protein expressions.

CONCLUSION

As confirmed by PCA and HMC-1 cell-based immunology assay, QE albumen and QE yolk may work together through exerting anti-allergy activity and can be used as a potential anti-allergic nutrient in the future.

摘要

背景

据报道鹌鹑蛋（QE）具有抗过敏和抗炎活性。我们已经证明，完整的鹌鹑蛋能够减轻食物过敏诱导的嗜酸性食管炎小鼠模型中的过敏症状，但鹌鹑蛋白或鹌鹑蛋黄是否起更重要的作用仍不清楚。

目的

在本研究中，我们研究了鹌鹑蛋在肥大细胞脱颗粒和效应期反应细胞因子产生中的抑制作用。

方法

采用被动皮肤过敏反应（PCA）小鼠模型来证实鹌鹑蛋的抗过敏作用。此外，使用HMC-1细胞模型更详细地研究其抑制作用。在本研究中，我们将鹌鹑蛋分为三组：完整鹌鹑蛋、鹌鹑蛋白和鹌鹑蛋黄。研究了鹌鹑蛋处理对肥大细胞脱颗粒和细胞内钙内流的影响。此外，还通过酶联免疫吸附测定（ELISA）、逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹法（western blotting）评估了鹌鹑蛋对过敏相关介质、基因和蛋白质的影响。

结果与讨论

我们的数据表明，用完整鹌鹑蛋（17mg/kg）处理的IgE介导的PCA小鼠中，肥大细胞脱颗粒介导的耳部血管通透性降低程度显著，降低了43.31±0.42%。基于HMC-1细胞的体外免疫学分析表明，鹌鹑蛋，特别是其蛋白，起到了“肥大细胞稳定剂”的作用。在70μg/mL的浓度下，鹌鹑蛋白有效地抑制了β-己糖胺酶、组胺和类胰蛋白酶的释放，以及Th2和促炎细胞因子的产生；降低了30%至50%。此外，鹌鹑蛋白还能够显著调节IL-10的上调，上调幅度达58.30±5.9%。有趣的是，我们的数据表明，鹌鹑蛋黄在其最高浓度（100μg/mL）时对调节Th2细胞因子仍有显著的抑制作用，而鹌鹑蛋白则没有抑制作用。蛋白质免疫印迹分析表明，鹌鹑蛋白有效地下调了钙相关蛋白（TRPC1、Orai1、STIM1、PLC-γ和IP3R）的表达，促进了PAR-2的减少，并诱导了JNK、IKKα、p50和p65蛋白表达的磷酸化减少。