细胞膜钙离子通路Orail的阻滞对实验性小鼠变应性鼻炎的影响

发布时间：2018-05-13 01:13

本文选题：Orai1 + 变应性鼻炎　；参考：《复旦大学》2010年博士论文

【摘要】： 目的：Orai1是组成钙离子释放激活的钙离子通道(CRAC通道)至关重要的亚单位,对存储调控性钙离子内流(SOCE)起着关键性作用。关于Orai1的研究目前已比较深入,但对于此通路在变应性鼻炎方面的作用目前尚未有研究涉及。本实验意欲检测Orai1在变应性鼻炎小鼠模型中的表达以及评估Orai1通路的阻滞对免疫球蛋白E (IgE)介导的过敏反应的影响。材料和方法：采用BALB/c品系小鼠,通过重复腹腔内注射卵清蛋白致敏、反复鼻腔内滴入卵清蛋白激惹建立变应性鼻炎小鼠模型,然后用免疫组化染色法、蛋白印迹法(western blotting)和实时反转录聚合酶链反应法(real-time RT-PCR)检测Orai1在对照组、实验组和2-氨基乙酯二苯基硼酸(2-APB)干预组小鼠鼻粘膜和鼻相关淋巴组织(NALT)中的表达,另外用酶联免疫吸附实验(ELISA)法分析上述几组小鼠鼻腔灌洗液中的白细胞三烯C4(LTC4)、嗜酸性粒细胞阳离子蛋白(ECP)、卵清蛋白特异性免疫球蛋白E (sIgE)和白细胞介素4(IL-4)的含量,并用real-time RT-PCR法检测几组小鼠鼻粘膜中的白细胞三烯C4合酶(LTC4S)信使核糖核酸(mRNA)、ECP mRNA的含量和小鼠NALT中的胚系Cε转录体、IL-4 mRNA的表达。结果：研究发现Orai1在鼻粘膜上皮细胞、粘膜下腺上皮细胞和NALT的免疫系统细胞中均有表达,在实验组表达尤为显著；2-APB鼻内滴入干预组小鼠相对于实验组小鼠的喷嚏数、挠鼻次数以及鼻粘膜内浸润的嗜酸性粒细胞数都明显减少,而且干预组Orai1在小鼠鼻粘膜和NALT中的表达也明显少于实验组,同时研究还发现,干预组小鼠鼻腔灌洗液中的LTC4、ECP、slgE和IL-4以及这些物质相应的mRNA的含量相对于实验组亦有显著抑制。结论：本实验证实Orai1在小鼠鼻粘膜和NALT中确有表达,2-APB小鼠鼻内滴入的干预措施可以有效地缓解小鼠变应性鼻炎的发作。
[Abstract]:Objective: Orai1 is a critical subunit of the CRAC channel, which is a calcium channel activated by calcium release. It plays a key role in the storage regulation of calcium ion influx (SOCEE). The research on Orai1 has been deeply studied, but the role of this pathway in allergic rhinitis has not been studied. The purpose of this study was to detect the expression of Orai1 in allergic rhinitis mice and to evaluate the effect of Orai1 pathway block on immunoglobulin E (IgE) -mediated allergic reaction. Materials and methods: the allergic rhinitis mouse model was established by repeated intraperitoneal injection of ovalbumin sensitized by repeated intraperitoneal injection of ovalbumin and repeated instillation of ovalbumin to induce allergic rhinitis in BALB/c strain mice. Western blotting and real-time RT-PCR were used to detect the expression of Orai1 in the nasal mucosa and naso-associated lymphoid tissue of mice in control group, experimental group and 2-APB intervention group. In addition, the contents of leukocyte triene C4Tc 4, eosinophil cationic protein (ECPP), ovalbumin specific immunoglobulin E (IgE) and interleukin-4 (IL-4) in the nasal lavage fluid of the above mentioned groups were analyzed by Elisa. The real-time RT-PCR method was used to detect the content of mRNA of leukocyte triene C4 synthase (LTC4S) in mouse nasal mucosa and the expression of IL-4 mRNA in mouse NALT. Results: the expression of Orai1 was found in nasal epithelial cells, submucosal gland epithelial cells and immune system cells of NALT, especially in experimental group. The number of nasal scratches and the number of eosinophils infiltrated in the nasal mucosa were significantly decreased, and the expression of Orai1 in the nasal mucosa and NALT in the intervention group was significantly lower than that in the experimental group. The contents of LTC4 ECPG E and IL-4 and their corresponding mRNA in nasal lavage fluid in the intervention group were also significantly inhibited compared with the experimental group. Conclusion: this study confirmed that the intervention of Orai1 expression in nasal mucosa and NALT could effectively relieve allergic rhinitis in mice.
【学位授予单位】：复旦大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R765.21

【共引文献】