Lin Lin1, Xia Zhao, Wenhong Yan, Weidong Qi. 1. Department of Otorhinolaryngology-Head and Neck Surgery, Huashan Hospital of Fudan University, Shanghai, China. linlinhs@yahoo.com.cn
Abstract
BACKGROUND: Orai1 is crucial for store-operated Ca2(+) entry and Ca2(+) release-activated Ca2(+) channel activities. However, little is known about its function in allergic diseases. OBJECTIVE: To assess the influence of Orai1 intervention on mouse airway epithelium reactions in vivo and in vitro. METHODS: We used immunohistochemical staining, enzyme-linked immunosorbent assay, and real-time reverse transcription-polymerase chain reaction to evaluate Orai1 expression in nasal and tracheal mucosa epithelium of nonsensitized, control, and 2-aminoethoxydiphenyl borate (2-APB)-treated groups in vivo and in vitro. In addition, we analyzed concentrations of interleukin 1β, interleukin 6, macrophage inflammatory protein 2, and tumor necrosis factor α in nasal lavage fluid, bronchoalveolar lavage fluid, and culture supernatant and their messenger RNAs in nasal and tracheal mucosa and cultured nasal and tracheal epithelium. RESULTS: Administration of 2-APB into the nostrils suppressed Orai1 expression in nasal and tracheal mucosa of treated mice compared with that in control mice and restrained the mediators in nasal lavage fluid, bronchoalveolar lavage fluid, and airway mucosa of treated groups compared with those in control groups. Similarly, the 2-APB intervention also alleviated Orai1 and the production of the mediators in culture supernatant and cultured airway epithelium under allergic conditions. CONCLUSIONS: Our results indicate that 2-APB could effectively ameliorate reactions of upper and lower airway epithelial cells in mice in allergic states in vivo and in vitro.
BACKGROUND:Orai1 is crucial for store-operated Ca2(+) entry and Ca2(+) release-activated Ca2(+) channel activities. However, little is known about its function in allergic diseases. OBJECTIVE: To assess the influence of Orai1 intervention on mouse airway epithelium reactions in vivo and in vitro. METHODS: We used immunohistochemical staining, enzyme-linked immunosorbent assay, and real-time reverse transcription-polymerase chain reaction to evaluate Orai1 expression in nasal and tracheal mucosa epithelium of nonsensitized, control, and 2-aminoethoxydiphenyl borate (2-APB)-treated groups in vivo and in vitro. In addition, we analyzed concentrations of interleukin 1β, interleukin 6, macrophage inflammatory protein 2, and tumor necrosis factor α in nasal lavage fluid, bronchoalveolar lavage fluid, and culture supernatant and their messenger RNAs in nasal and tracheal mucosa and cultured nasal and tracheal epithelium. RESULTS: Administration of 2-APB into the nostrils suppressed Orai1 expression in nasal and tracheal mucosa of treated mice compared with that in control mice and restrained the mediators in nasal lavage fluid, bronchoalveolar lavage fluid, and airway mucosa of treated groups compared with those in control groups. Similarly, the 2-APB intervention also alleviated Orai1 and the production of the mediators in culture supernatant and cultured airway epithelium under allergic conditions. CONCLUSIONS: Our results indicate that 2-APB could effectively ameliorate reactions of upper and lower airway epithelial cells in mice in allergic states in vivo and in vitro.