重组干扰素γ对变应性鼻炎大鼠鼻黏膜中NLRP3炎症小体活化的影响  

Effect of recombinant interferon-gamma on activation of NLRP3 inflammasome in nasal mucosa of rats with allergic rhinitis

作　　者：李儒佑[1] 冯六连[1] 柯正华[1] 周朕[1] 黄永军 王钢胜 LI Ruyou;FENG Liulian;KE Zhenghua;ZHOU Zhen;HUANG Yongjun;WANG Gangsheng(Department of Respiratory Medicine,Edong Medical Group Huangshi Central Hospital,Affiliated Hospital of Hubei Polytechnic University,Huangshi 435000,China)

机构地区：[1]鄂东医疗集团黄石市中心医院湖北理工学院附属医院呼吸内科

出　　处：《山西医科大学学报》2019年第9期1305-1309,共5页Journal of Shanxi Medical University

基　　金：湖北省自然科学基金资助项目(WJ2015MB278)

摘　　要：目的 探讨重组干扰素γ(IFN-γ)对变应性鼻炎(AR)大鼠鼻黏膜中NLRP3炎症小体活化的影响。方法 8周龄雄性SD大鼠30只,随机分为对照组、AR组和IFN-γ组,每组10只。卵清蛋白(OVA)法制备大鼠AR模型,AR组和IFN-γ组大鼠从激发第1天开始,在给予OVA激发前30 min,分别用PBS和IFN-γ滴鼻,50 μl/侧,1次/d。连续激发20 d,末次激发后30 min内,对大鼠行为学评分;各组大鼠于评价结束后,RT-PCR检测大鼠鼻黏膜中NLRP3、ASC和Caspase-1 mRNA表达,Western blot检测大鼠鼻黏膜中NLRP3、ASC、pro-Caspase-1和Caspase-1 P20蛋白表达,ELISA法检测大鼠双侧鼻腔灌洗液中IL-1β、IL-6和IL-8水平及血清中OVA-sIgE浓度。结果 成功构建了AR大鼠模型。与AR组比较,重组IFN-γ滴鼻可降低AR大鼠行为学评分( P <0.01);AR组大鼠鼻黏膜中NLRP3、ASC和Caspase-1 mRNA相对表达量均高于对照组( P <0.05),NLRP3、ASC和Caspase-1 P20蛋白相对表达量均高于对照组,而pro-Caspase-1蛋白相对表达量低于对照组( P <0.05);重组IFN-γ滴鼻大鼠鼻黏膜中NLRP3、ASC和Caspase-1 mRNA表达量均明显低于AR组( P <0.05),NLRP3、ASC、Caspase-1和Caspase-1 P20蛋白相对表达量均低于AR组,而Pro-Caspase-1蛋白相对表达量高于AR组( P <0.05);AR组大鼠双侧鼻腔灌洗液中IL-1β、IL-6和IL-8水平及血清中OVA-sIgE浓度均高于对照组( P <0.05),重组IFN-γ滴鼻大鼠双侧鼻腔灌洗液中IL-1β、IL-6和IL-8水平及血清中OVA-sIgE浓度均明显低于AR组( P <0.05)。结论 重组IFN-γ滴鼻可影响NLRP3炎症小体介导的炎症反应,从而减轻AR大鼠变应性鼻炎症状。Objective To investigate the effect of IFN-γ on activation of NLRP3 inflammasome in nasal mucosa of rats with allergic rhinitis(AR). Methods Thirty male SD rats aged 8 weeks were randomly divided into control group, AR group and IFN-γ group, with 10 rats in each group. Rat AR model was prepared using ovalbumin(OVA). The rats in AR group and IFN-γ group were nasally given phosphate buffered saline(PBS) and IFN-γ 50 μl/side, once a day, from the 1st day of challenge and 30 min before the OVA challenge, respectively. After continuous excitation for 20 d, the behavioral scores of rats were evaluated within 30 min after the last challenge. After the evaluation, the expression levels of NLRP3, ASC and Caspase-1 mRNA in nasal mucosa of rats were detected by real-time fluorescent quantitative PCR. The expression levels of NLRP3, ASC, pro-Caspase-1 and Caspase-1 P20 proteins in rat nasal mucosa were detected by Western blot. The concentrations of IL-1β, IL-6 and IL-8 in bilateral nasal lavage fluid and OVA-sIgE in serum were detected by ELISA. The AR rat model was successfully constructed. Results Compared with AR group, recombinant IFN-γ nasal drops reduced the behavioral score of AR rats( P <0.01). The expression levels of NLRP3, ASC and Caspase-1 mRNA in the nasal mucosa of rats in AR group were higher than those in control group( P <0.05). The expression levels of NLRP3, ASC and Caspase-1 P20 proteins were higher in AR group than in control group, while the expression level of Pro-Caspase-1 protein was lower( P <0.05). After recombinant IFN-γ nasal drops, the expression levels of NLRP3, ASC and Caspase-1 mRNA and proteins in the nasal mucosa of rats were significantly lower in IFN-γ group than in AR group( P <0.05). The expression levels of NLRP3, ASC, Caspase-1 and Caspase-1 P20 proteins were lower in IFN-γ group than in AR group, while the expression level of pro-Caspase-1 protein was higher( P <0.05). The concentrations of IL-1β, IL-6 and IL-8 in bilateral nasal lavage fluid and OVA-sIgE in serum of rats

关 键 词：变应性鼻炎 干扰素Γ NLRP3炎症小体 大鼠 

分 类 号：R765.21[医药卫生—耳鼻咽喉科]