Department of Otolaryngology-Head and Neck Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, PR China; Otolaryngological Institute, Shanghai Jiao Tong University, Shanghai, PR China; Shanghai Key Laboratory of Sleep Disordered Breathing, Shanghai, PR China.
Department of Otolaryngology-Head and Neck Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, PR China; Otolaryngological Institute, Shanghai Jiao Tong University, Shanghai, PR China; Shanghai Key Laboratory of Sleep Disordered Breathing, Shanghai, PR China.
Int Immunopharmacol. 2020 Jun;83:106394. doi: 10.1016/j.intimp.2020.106394. Epub 2020 Mar 16.
Allergic rhinitis (AR) is a complex IgE-mediated nasal allergic and inflammatory disease. Nucleotide-binding domain (NOD)-like receptor protein 3 (NLRP3) is essential in the process of allergic and inflammatory responses. MCC950 is a selective NLRP3 inhibitor. However, its role and mechanism in AR remains undetermined. The present study aimed to explore the effect and mechanism of MCC950 on an ovalbumin (OVA) induced mouse model of AR. The AR BALB/c mice were constructed using OVA and administrated intranasally with MCC950. Concentrations of OVA-specific IgE, histamines and leukotrienes C4 (LTC4) in serum, and OVA-specific IgE, ECP, IFN-γ, IL-4, IL-5, IL-13, IL-1β and IL-18 in nasal lavage fluid (NLF) were assayed by enzyme-linked immunosorbent assay (ELISA). Inflammatory cells were counted in NLF. HE and PAS staing were used for evaluating eosinophils and goblet cells. Immunohistochemistry (IHC) staining were employed to evaluate immunolabeling of NLRP3, Caspase-1, ASC, IL-1β and IL-18 in nasal mucosas of mice. Real-time PCR was conducted to assay NLRP3, Caspase-1, ASC, IL-1β and IL-18 mRNA levels. In vitro studies, western blotting, real-time PCR and ELISA were performed to evaluate the effects and mechanisms of OVA and NLRP3 inhibitor MCC950 on spleen mononuclear cells. We found significant downregulation of sneezing, nasal rubbing, inflammatory cytokines, inflammatory cells and NLRP3, Caspase-1, ASC, IL-1β and IL-18 expression in MCC950 treated mice compared with untreated AR mice. In spleen mononuclear cells culture and stimulation experiment, NLRP3, Caspase-1, ASC, IL-1β and IL-18 levels were upregulated by OVA but inhibited by MCC950. In conclusion, MCC950 could effectively exert its ameliorative effect in murine AR by inhibiting NLRP3 and leads to reduction of Caspase-1, ASC, IL-1β and IL-18, resulting in the attenuation of the allergic and inflammatory responses.
变应性鼻炎（AR）是一种复杂的 IgE 介导的鼻过敏和炎症性疾病。核苷酸结合域（NOD）样受体蛋白 3（NLRP3）是过敏和炎症反应过程中的必需物质。MCC950 是一种选择性 NLRP3 抑制剂。然而，其在 AR 中的作用和机制尚不清楚。本研究旨在探讨 MCC950 对卵清蛋白（OVA）诱导的 AR 小鼠模型的作用及其机制。采用 OVA 构建 AR BALB/c 小鼠模型，并鼻内给予 MCC950 进行干预。通过酶联免疫吸附试验（ELISA）检测血清中 OVA 特异性 IgE、组胺和白三烯 C4（LTC4）的浓度，以及鼻洗液（NLF）中 OVA 特异性 IgE、ECP、IFN-γ、IL-4、IL-5、IL-13、IL-1β 和 IL-18 的浓度。通过计数 NLF 中的炎性细胞来评估炎症细胞。采用 HE 和 PAS 染色评估嗜酸性粒细胞和杯状细胞。采用免疫组织化学（IHC）染色评估小鼠鼻黏膜中 NLRP3、Caspase-1、ASC、IL-1β 和 IL-18 的免疫标记。通过实时 PCR 检测 NLRP3、Caspase-1、ASC、IL-1β 和 IL-18 mRNA 水平。进行体外研究，采用 Western blot、实时 PCR 和 ELISA 检测 OVA 和 NLRP3 抑制剂 MCC950 对脾单核细胞的作用及其机制。结果发现，与未经治疗的 AR 小鼠相比，MCC950 治疗组小鼠的喷嚏、鼻擦、炎性细胞因子、炎性细胞和 NLRP3、Caspase-1、ASC、IL-1β 和 IL-18 表达均显著下调。在脾单核细胞培养和刺激实验中，OVA 可上调 NLRP3、Caspase-1、ASC、IL-1β 和 IL-18 水平，而 MCC950 可抑制其表达。综上所述，MCC950 通过抑制 NLRP3 有效发挥其对 AR 小鼠的治疗作用，导致 Caspase-1、ASC、IL-1β 和 IL-18 减少，从而减轻过敏和炎症反应。
