Insights in Clinical and Cellular Immunology

ISSN:2640-2793

Research Article

Effect of common food additives on mast cell activation

Carena MP，Mariani ML，OrdóñezA和Penissi AB*

Instituto deHastorologíaymbriología“博士马里奥·H·布尔戈斯（Mario H. Burgos

*Address for Correspondence:Alicia Beatriz Penissi，教授，博士学位，Instituto deHestorologíayEmbriología“博士Mario H. Burgos”（Ihem-Conicet），ciencias a ciencias ersivard de Cuyo大学。Casilla de Correo 56.（5500）Mendoza，阿根廷，电话：00-54-0261-4135000;2670/2672/2673;传真：00-54-261-4494117;电子邮件：apenissi@fcm.uncu.edu.ar

Dates:Submitted:03 January 2019;Approved:16 January 2019;Published:2019年1月17日

如何引用本文：Carena MP，Mariani ML，OrdóñezA，Penissi AB。常见食物添加剂对肥大细胞激活的影响。见解临床细胞免疫。2019;3：001-005。doi：10.29328/journal.icci.1001007

版权许可证：©2019 Carena MP等。这是根据Creativ金博宝app体育e Commons归因许可分发的开放访问文章，该文章允许在任何媒介中不受限制地使用，分发和复制，前提是适当地引用了原始作品。

肥大细胞是专门的结缔组织细胞，在过敏和炎症反应的起源和调节中起着核心作用[1-4]。

考虑到允许主要食品添加剂的一些作者描述的过敏性特性（Tartrazine，Bisuimbhite和Benzoate钠）[5-7]，我们的工作假设表明，这些食物添加剂刺激了肥大细胞的脱粒化。

本工作的总体目的是研究肥大细胞与批准用于食品的最常见添加剂之间的相互作用。

分析化合物

The food additives tartrazine, sodium benzoate and sodium bisulphite, as well as the calcium ionophore A23187 were purchased from Sigma.

Animals

Male Wistar adult rats weighing approximately 300 to 500 g, infection free and maintained under a 12-hour dark/light cycle in a temperature-controlled room (24-25ºC) with free access to drinking water and laboratory food, were used for the study. All animal experiments were carried out according to the standards included in the Guide for the Care and Use of Laboratory Animals (published by the National Academy of Science, National Academy Press, Washington, D.C.), and approved by the Institutional Committee for Care and Use of Laboratory Animals (CICUAL, Facultad de Ciencias Médicas, Universidad Nacional de Cuyo, Mendoza, Argentina).

大鼠肥大细胞的分离和纯化

Mast cells were obtained by peritoneal lavage with physiological solution and purified in a discontinuous gradient of sterile Percoll solution. The purity of the cells was evaluated by staining with 1% toluidine blue. For the tests, preparations with purity of 95-100% were used. The viability of the mast cells was evaluated with trypan blue.

一般协议

The purified mast cells were incubated with: 1) buffer solution or 2) stimulus. The stimuli were tartrazine, sodium benzoate, sodium bisulphite and the calcium ionophore A23187. A23187 was used as a reference mast cell secretagogue. Different doses and combinations of food additives were used (these doses include those commonly used in food and those referred by the literature). The viability of the mast cells was evaluated with trypan blue. In the incubation solutions, the release of β-hexosaminidase was quantified by colorimetric reaction and ELISA plate reader (the release of this enzyme is used as a mast cell degranulation marker). The remaining β-hexosaminidase concentration (not released) was studied in the cells after the incubations. Morphology of the mast cells was analyzed by light microscopy after experiments.

Quantification of β-hexosaminidase

在孵育溶液和其余细胞中，β-己糖胺酶的水平通过比色反应和ELISA板读取器测量。

Light microscopy with blue toluidine stain

将肥大细胞固定在2％的戊二醛中。在固定剂中2小时后，将细胞悬浮液用甲苯胺蓝（0.1％W/V，pH 3.0）染色，放置在载玻片和覆盖面之间，并在Olympus CH-2显微镜下进行检查。在放大倍率的放大倍率上定量表现出脱粒的肥大细胞的百分比。肥大细胞的激活定义为挤出的颗粒的存在靠近所讨论的细胞表面，或用甲苯胺蓝的一半或更少的细胞染色。

统计处理

The results were analyzed using analysis of variance type 1 (ANOVA-1) followed by the Tukey-Kramer test. P <0.05 was considered statistically significant.

The food additives tartrazine, sodium benzoate and sodium bisulphite did not stimulate the release of β-hexosaminidase from mast cells at any of the concentrations used (Figures 1-4). In contrast, tartrazine at concentrations of 0.1 μM and 1 μM, and sodium benzoate and sodium bisulphite at concentrations of 0.1 μM, 1 μM, 10 μM and 100 μM, significantly inhibited the basal release of β-hexosaminidase from mast cells (Figures 1-4). Higher concentrations of the additives did not inhibit this release, probably due to cell death.

食物添加剂的抑制作用不伴随细胞活力的变化。锥虫蓝排放测试表明，在抑制浓度下的生存力大于80％，在非抑制剂量下的生存力低于80％（数据未显示）。

考虑到某些常用的食物添加剂（酸酸盐，苯甲酸钠和亚硫酸钠浓度为0.1μm，1μm）没有刺激，但抑制了腹膜桅杆细胞的脱粒，我们决定确定这些添加剂对脱粒的影响由钙离子载体A23187诱导的肥大细胞。在该实验模型中，亚硫酸钠（单独并与苯甲酸钠和/或亚硫酸钠结合）抑制了由钙离子载体A23187诱导的肥大细胞活化。获得的结果如图5所示。

我们关于β-己糖胺酶释放的生化结果与光和显微镜获得的结果一致（图6）。

The present study shows that the commonly used food additives (tartrazine, sodium benzoate and sodium bisulphite) do not stimulate basal degranulation of mast cells in an in vitro model of peritoneal mast cells. By the contrary, these additives inhibit the basal release of β-hexosaminidase from mast cells at low concentrations. This means that some physiological effects of mast cells could be affected by these food additives.

However, sodium bisulphite inhibits mast cell activation induced by the calcium ionophore A23187 in this same experimental model. It is the first time that a stabilizing action at mast cell level has been demonstrated for sodium bisulphite.

亚硫酸钠抑制作用的机理与该化合物与羰基化合物反应并抑制FcεRI受体的能力有关[8,9]。但是，需要进一步的研究来解释这些作用的确切分子机制。

该化合物在治疗和 /或预防由肥大细胞介导的病理以及营养生物化学领域的病理方面可以代表有趣的替代方法。