IL-17A激活NLRP3炎性小体在CRSwNP中的作用及临床意义  

Effects and clinical significance of NLRP3 inflammasome activated by IL-17A in CRSwNP

作　　者：李越 张雅娜 常利红[1] 黄伟强 吴浩天 吴喜福[1] 黄子真[1] 张革化[1] Li Yue;Zhang Yana;Chang Lihong;Huang Weiqiang;Wu Haotian;Wu Xifu;Huang Zizhen;Zhang Gehua(Department of Otorhinolaryngology Head and Neck Surgery,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China)

机构地区：[1]中山大学附属第三医院耳鼻咽喉头颈外科,广州510630

出　　处：《中华耳鼻咽喉头颈外科杂志》2023年第7期690-698,共9页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：国家自然科学基金(81970859);广东省自然科学基金面上项目(2019A1515011093);广州市科技计划项目(202002030034)。

摘　　要：目的探讨慢性鼻窦炎伴鼻息肉(CRSwNP)中白细胞介素(IL)-17A激活NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor family pyrin domain containing 3,NLRP3)的作用及临床意义。方法收集自2020年1月至2021年12月期间在中山大学附属第三医院接受鼻内镜手术治疗的患者标本,其中CRSwNP患者28例(男性19例,女性9例,年龄19~67岁),慢性鼻窦炎不伴鼻息肉(CRSsNP)患者22例(男性14例,女性8例,年龄26~66岁),对照组患者22例(男性13例,女性9例,年龄20~57岁)。实时定量聚合酶链反应(qRT-PCR)检测IL-17A、NLRP3、IL-1β及IL-18在3组中的表达,分析其相关性;组织免疫荧光三染定位分析IL-17A、NLRP3、IL-18在鼻黏膜组织中的位置;蛋白免疫印迹法及酶联免疫吸附试验检测IL-17A刺激原代鼻黏膜上皮细胞及使用IL-17A受体抑制剂后NLRP3、IL-1β及IL-18在细胞中的表达;细胞免疫荧光观察IL-17A刺激原代鼻黏膜上皮细胞后,NLRP3、IL-1β、IL-18蛋白的表达,以及使用IL-17A受体抑制剂和NLRP3抑制剂(MCC950)后,IL-17A刺激原代鼻黏膜上皮细胞NLRP3、IL-1β、IL-18蛋白的表达。分析NLRP3、IL-1β及IL-18与CRSwNP患者CT评分、鼻内镜评分、视觉模拟量表(VAS)评分和鼻腔鼻窦结局测试(SNOT)22评分之间的相关性。采用SPSS 20.0软件对数据进行统计。结果CRSwNP组IL-17A、NLRP3、IL-1β及IL-18 mRNA表达明显高于CRSsNP组(P=0.018,P<0.001,P=0.005,P<0.016)及对照组(P值均<0.001),且IL-17A与NLRP3、IL-1β、IL-18的表达存在正相关(r值分别为0.643、0.650、0.629,P值均<0.05)。息肉组织中,IL-17A与NLRP3、IL-18共同定位于上皮层。IL-17A刺激原代鼻黏膜上皮细胞后,NLRP3、IL-1β、IL-18表达增高;使用IL-17A受体抑制剂后,可明显下调NLRP3、IL-1β、IL-18的表达;使用NLRP3抑制剂MCC950后则明显下调IL-17A促进NLRP3、IL-1β及IL-18表达的作用。NLRP3、IL-1β、IL-18的表达与CRSwNP患者CT、鼻内镜、VAS和SNOT22评分存在正相关。结论CRSwNObjective To investigate the effects and clinical significance of NOD-like receptor family pyrin domain containing 3(NLRP3)inflammasome activated by interleukin(IL)-17A in chronic rhinosinusitis with nasal polyps(CRSwNP).MethodsPatients underwent nasal endoscopic surgery in the Third Affiliated Hospital of Sun Yat-sen University from January 2020 to December 2021 were collected,including 28 CRSwNP(including 19 males and 9 females,aged 19 to 67 years),22 chronic rhinosinusitis without nasal polyps(CRSsNP)and 22 controls.qRT-PCR was used to detect the expressions of IL-17A,NLRP3,IL-1βand IL-18 in the three groups,and their correlations were analyzed.The positions of IL-17A,NLRP3 and IL-18 in nasal polys were analyzed by immunofluorescence.Western Blotting and ELISA were employed to detect the expression of NLRP3,IL-1βand IL-18 in the human nasal epithelial cells after using IL-17A stimulation or IL-17A receptor inhibitor.Immunofluorescence was used to observe the NLRP3,IL-1β,and IL-18 protein expression after IL-17A stimulating human nasal epithelial cells,and after the use of IL-17A receptor inhibitor and NLRP3 inhibitor MCC950.The correlations between NLRP3,IL-1β,IL-18 and CT scores,nasal endoscopic scores,visual analogue scale(VAS)scores,and sino-nasal outcome test(SNOT)22 scores of CRSwNP patients were analyzed.SPSS 20.0 software was used for statistical analysis.ResultsThe expressions of IL-17A,NLRP3,IL-1βand IL-18 in the tissues of CRSwNP patients were significantly higher than those in CRSsNP group(P=0.018,P<0.001,P=0.005,P=0.016)and the control group(all P<0.001).IL-17A was positively correlated with the expression of NLRP3,IL-1β,and IL-18(r ralue was 0.643,0.650,0.629,respectively,all P<0.05).IL-17A,NLRP3,and IL-18 were co-localized in the epithelial propria of polyp tissue.IL-17A stimulated the expressions of NLRP3,IL-1β,and IL-18 in human nasal epithelial cells.After the use of IL-17A receptor inhibitor,the expressions of NLRP3,IL-1β,and IL-18 were significantly down-regulated.After the use of NL

关 键 词：鼻窦炎 鼻息肉 IL-17A NLRP3炎性小体 

分 类 号：R765.9[医药卫生—耳鼻咽喉科]