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

Objective: 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). Methods: Patients 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-1β and 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-1β and 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. Results: The expressions of IL-17A, NLRP3, IL-1β and 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 NLRP3 inhibitor MCC950, IL-17A was significantly down-regulated to promote the expression of NLRP3, IL-1β, and IL-18. The expressions of NLRP3, IL-1β and IL-18 were positively correlated with CT, nasal endoscopy, VAS, and SNOT22 scores in patients with CRSwNP. Conclusions: IL-17A promotes the release of IL-1β and IL-18 by activating the NLRP3 inflammasome and aggravates the severity of the disease in CRSwNP.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： CRSwNP中，IL-17A可通过激活NLRP3炎性小体，促进IL-1β及IL-18的释放，加重疾病的严重程度。.