Shi Le, Ma Yue, Zheng Chunquan, Zhang Qingzhao
Department of Otorhinolaryngology, Eye & Ear Nose and Throat Hospital, Fudan University, Shanghai, 200031, People's Republic of China.
Department of Otorhinolaryngology, Eye & Ear Nose and Throat Hospital, Fudan University, Shanghai, 200031, People's Republic of China.
Int J Pediatr Otorhinolaryngol. 2017 Jul;98:32-38. doi: 10.1016/j.ijporl.2017.04.036. Epub 2017 Apr 24.
This study aimed to investigate the effect of blocking Notch signaling by γ-secretase inhibitor (GSI) on allergic rhinitis.
GSI, N-[N-(3,5-difluorophenacetyl-L-alanyl)]-S-phenylglycine t-butylester (DAPT) was administered to ovalbumin-induced AR mice models intranasally. We observed symptoms of sneezing and nose rubbing. To detect the inflammatory state, the serum OVA-specific-IgE, IFN-γ, IL-4, and IL-5 were analyzed by ELISA, and Th cell cytokines in nasal mucosa were analyzed by RT-PCR, including T-bet, IFN-γ, GATA-3, IL-4, and IL-5. In addition, hematoxylin-eosin (HE) and periodic acid-Schiff (PAS) were applied for histopathological examination. As for the evaluation of Notch signaling, we analyzed the Notch-1, Notch signaling target Hes-1, and Hes-5 in mucosa by RT-PCR, besides, used western blotting and immunohistochemistry to assess NICD (Notch intracellular domain).
The results showed that the DAPT ameliorated the development of AR and suppressed Th2 cytokine levels significantly, alleviating eosinophils infiltration and goblet cells metaplasia, suggesting that the GSI can regulate Th2 response and weaken airway inflammation in AR.
Our findings provide evidence that blocking Notch signaling by GSI offers high value in treating AR.
本研究旨在探讨γ-分泌酶抑制剂（GSI）阻断Notch信号通路对变应性鼻炎的影响。
将GSI，N-[N-(3,5-二氟苯乙酰基-L-丙氨酰)]-S-苯基甘氨酸叔丁酯（DAPT）经鼻给予卵清蛋白诱导的变应性鼻炎小鼠模型。观察打喷嚏和擦鼻症状。为检测炎症状态，采用酶联免疫吸附测定法分析血清卵清蛋白特异性免疫球蛋白E、干扰素-γ、白细胞介素-4和白细胞介素-5，并采用逆转录-聚合酶链反应分析鼻黏膜中的辅助性T细胞细胞因子，包括T盒转录因子、干扰素-γ、GATA结合蛋白3、白细胞介素-4和白细胞介素-5。此外，采用苏木精-伊红染色和过碘酸-希夫染色进行组织病理学检查。至于Notch信号通路的评估，我们通过逆转录-聚合酶链反应分析黏膜中的Notch-1、Notch信号通路靶点Hes-1和Hes-5，此外，还采用蛋白质免疫印迹法和免疫组织化学法评估Notch胞内结构域（NICD）。
结果显示，DAPT改善了变应性鼻炎的发展，并显著抑制了辅助性T2细胞因子水平，减轻了嗜酸性粒细胞浸润和杯状细胞化生，表明GSI可调节辅助性T2细胞反应并减轻变应性鼻炎中的气道炎症。
我们的研究结果提供了证据，证明GSI阻断Notch信号通路在治疗变应性鼻炎方面具有很高的价值。
