花生过敏小鼠腹腔肥大细胞SOC活性与ROS相关性研究

The Experiment Study of Peritoneal Mast Cell SOC Activity of Mice That Have Been Therapied by Antioxidant

研究花生过敏状态下的C57BL/6小鼠腹腔肥大细胞的激活与钙离子通道之间的内在联系,以及探讨肥大细胞钙离子通道与ROS(reactive oxygen species)之间的关系.18只雌性C57BL/6小鼠随机分为PBS组(A组)、花生过敏模型组(B组)、vitamin A和vitamin E灌胃治疗组(C组).酶联免疫吸附实验检测各组腹腔上清液中特异性的IgE和IgG2a水平,以及细胞因子的变化,另外测定腹腔上清液与血清中的组胺、ROS含量.腹腔灌洗液中细胞涂片,进行肥大细胞计数和脱颗粒比例统计,荧光共聚焦显微镜观察腹腔肥大细胞钙离子通道SOC(Store oprated channel)的活性.腹腔上清液中花生特异性IgE、IL-4、IL-10、组胺、ROS、血清中的组胺和ROS、腹腔灌洗液中肥大细胞总数、肥大细胞脱颗粒比例,都呈现出A组<C组<B组的趋势.腹腔上清液中IgG2a、1L-6、IL-10的水平B组低于A组,但高于C组.共聚焦显微镜观察肥大细胞内部钙离子荧光强度数据表明B组小鼠腹腔肥大细胞SOC通道有更强的生理活性,而C组肥大细胞SOC通道被抑制,表明ROS的水平与肥大细胞ROS活性是正相关.花生过敏组肥大细胞SOC较PBS组活性更高,抗氧化剂药物可能通过清除ROS来抑制肥大细胞SOC通道的活性,进而抑制肥大细胞脱颗粒,缓解过敏症状.这为进一步研究抗氧化剂治疗花生过敏反应的机理奠定了基础,同时对临床治疗过敏反应也有一定的指导意义.

By establish mice peanut allergy and antioxidant therapy allergy mice model, to investigated correlation ac- tivation of mast cell with SOC channel, and further to study the relationship between mast cell SOC and ROS. 18 fe- male C57BL/6 mice were divided into three groups that one is treated by PBS and the other is treated by lavaging crude peanut protein, another is peanut allergy mice that been therapied by lavaging vitamin A and vilamin E. Peanut protein specific IgE and IgG2a, cytokines, histamin and ROS in supernatant of peritoneal lavaging were assayed by en- zyme-linked immunoassay（ELISA）. In addition, hiatamin and ROS in blood serum were also assayed by ELISA. By peritoneal lavaging, inflammatory cells and mast cell were anlalyzed. Confocal was used to detect SOC activity of peritoneal mast cell. In peritoneal lavaging experiment, the total numbers of cells, eosinophil, mast cell of B group were greatly increased than A group, but these numbers of C groups were lower than B group. The levels of IgE, IL-4,IL-10 in supernatant of peritoneal lavaging liquid were presented a trendency, which C group were lower than B group, but higher than A group. But the levels of IgG2a, 1L-6,IL-10 were differences, which C group were lower than A group, but higher than B group. The levels of histamin and ROS presented a trendency, which C group were lower than B group, but higher than A group. Confocal recorded the fluorescence intensity of intracellular calcium changes. The datas of confocat recorded show that SOC activity of B groups mast cells were increased, but contrast to B group, SOC activity of C group were inhibited. Vitamin A and vitamin C can relieve allergy induced by peanut crud protein. However anti- oxidant can inhibit SOC activity of mast cell by scavenge ROS. Accordingly antioxidant depressed mast cell degranula- tion and then relieve anaphylactic response. This study laid the foundation for further research in the mechanism of an- tioxidants treatment of peanut allergy, while it has some significance for the clinical treatment of anaphylaxis.