Ameliorative effect of selective NLRP3 inflammasome inhibitor MCC950 in an ovalbumin-induced allergic rhinitis murine model
Ameliorative effect of selective NLRP3 inflammasome inhibitor MCC950 in an ovalbumin-induced allergic rhinitis murine model
复制标题
选择性NLRP3炎症小体抑制剂MCC950对卵清蛋白诱导的过敏性鼻炎小鼠模型的改善作用
DOI：
10.1016/j.intimp.2020.106394
复制
发表时间：
2020-06-01
影响因子：
5.6
通讯作者：
Lin, Hai
中科院分区：
文献类型：
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作者：
Zhang, Weitian;Ba, Guangyi;Lin, Hai
变应性鼻炎（AR）是一种复杂的ige介导的鼻腔变应性炎症性疾病。核苷酸结合域（NOD）样受体蛋白3 （NLRP3）在过敏和炎症反应过程中至关重要。MCC950是一种选择性NLRP3抑制剂。然而，其在AR中的作用和机制尚未确定。本研究旨在探讨MCC950对卵清蛋白（OVA）诱导的AR小鼠模型的影响及其机制。采用OVA构建AR BALB/c小鼠，经鼻给药MCC950。采用酶联免疫吸附法（ELISA）检测ova特异性IgE、组胺、白三烯C4 （LTC4）及ova特异性IgE、ECP、ifn - γ、IL-4、IL-5、IL-13、IL-1 β和IL-18在鼻腔灌洗液（NLF）中的浓度。NLF中计数炎性细胞。HE和PAS染色用于评价嗜酸性粒细胞和杯状细胞。采用免疫组化（IHC）染色法评价小鼠鼻黏膜NLRP3、Caspase-1、ASC、IL-1 β和IL-18的免疫标记。Real-time PCR检测NLRP3、Caspase-1、ASC、IL-1 β和IL-18 mRNA水平。体外实验采用western blotting、real-time PCR和ELISA检测OVA和NLRP3抑制剂MCC950对脾单核细胞的影响及机制。我们发现，与未治疗的AR小鼠相比，MCC950治疗小鼠打喷嚏、擦鼻、炎症细胞因子、炎症细胞和NLRP3、Caspase-1、ASC、IL-1 β和IL-18的表达显著下调。在脾单核细胞培养和刺激实验中，卵细胞可上调NLRP3、Caspase-1、ASC、IL-1 β和IL-18水平，MCC950可抑制其表达。综上所述，MCC950可通过抑制NLRP3有效发挥其对小鼠AR的改善作用，导致Caspase-1、ASC、IL-1 β和IL-18的减少，从而减弱过敏和炎症反应。
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-gamma, IL-4, IL-5, IL-13, IL-1 beta 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-1 beta and IL-18 in nasal mucosas of mice. Real-time PCR was conducted to assay NLRP3, Caspase-1, ASC, IL-1 beta 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-1 beta 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-1 beta 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-1 beta and IL-18, resulting in the attenuation of the allergic and inflammatory responses.