Department of Otolaryngology-Head and Neck Surgery, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.
Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, Ill.
J Allergy Clin Immunol. 2022 Aug;150(2):337-351. doi: 10.1016/j.jaci.2022.02.031. Epub 2022 Mar 26.
Pyroptosis is closely related to inflammation. However, the molecular mechanisms and pathologic contributions of pyroptotic epithelial cell are not yet fully understood.
This study aimed to explore the function and molecular mechanisms of IL-17A on human nasal epithelial cell (hNEC) pyroptosis.
The expression of pyroptosis-related biomarkers and IL-17A was assessed in sinonasal mucosa from control individuals, patients with chronic rhinosinusitis without nasal polyps, and patients with chronic rhinosinusitis with nasal polyps (CRSwNP) by using quantitative RT-PCR. Their localization was analyzed via immunohistochemistry and immunofluorescence. The ultrastructural characteristics of IL-17A-induced pyroptosis in hNECs were visualized by using electron microscopy. IL-17A functional assays were performed on hNECs and airway epithelial cell lines. Cytokine levels were quantified via ELISA. The signaling pathways involved in IL-17A-induced pyroptosis were studied via unbiased RNA sequencing and Western blotting.
The expression of IL-17A and the pyroptotic biomarkers NOD-like receptor family, pyrin domain containing 3 (NLRP3), caspase-1, gasdermin D, and IL-1β was increased in nasal mucosa from patients with CRSwNP compared with in those with chronic rhinosinusitis without nasal polyps and the control subjects. IL-17A was positively correlated and colocalized with the pyroptotic biomarkers. IL-17A treatment induced pyroptosis in the hNECs and cell lines analyzed, primarily through the extracellular signal-regulated kinase (ERK)-NLRP3/caspase-1 signaling pathway, and increased IL-1β and IL-18 secretion in hNECs. Moreover, IL-17A-induced pyroptosis contributed to steroid resistance by affecting glucocorticoid receptor-α and glucocorticoid receptor-β expression, and the inhibition of pyroptotic proteins partially abolished IL-17A-induced steroid resistance in hNECs.
Elevated IL-17A level promotes pyroptosis in hNECs through the ERK-NLRP3/caspase-1 signaling pathway and contributes to glucocorticoid resistance by affecting glucocorticoid receptor homeostasis in patients with CRSwNP.
细胞焦亡与炎症密切相关。然而,细胞焦亡的上皮细胞的分子机制和病理贡献尚不完全清楚。
本研究旨在探讨白细胞介素 17A(IL-17A)对人鼻上皮细胞(hNEC)细胞焦亡的作用及其分子机制。
采用实时定量 RT-PCR 检测对照组、慢性鼻-鼻窦炎伴鼻息肉(CRSwNP)患者和慢性鼻-鼻窦炎不伴鼻息肉(CRSsNP)患者鼻黏膜中细胞焦亡相关标志物和 IL-17A 的表达,免疫组织化学和免疫荧光法分析其定位,电子显微镜观察 IL-17A 诱导 hNEC 细胞焦亡的超微结构特征。采用 hNEC 和气道上皮细胞系进行 IL-17A 功能实验,酶联免疫吸附试验检测细胞因子水平。采用无偏 RNA 测序和 Western blot 研究 IL-17A 诱导细胞焦亡涉及的信号通路。
与 CRSsNP 患者相比,CRSwNP 患者鼻黏膜中 IL-17A 和细胞焦亡标志物 NOD 样受体家族、含pyrin 结构域蛋白 3(NLRP3)、半胱天冬酶 1、Gasdermin D 和白细胞介素 1β的表达增加。IL-17A 与细胞焦亡标志物呈正相关且共定位。在本研究分析的 hNEC 和细胞系中,IL-17A 诱导细胞焦亡,主要通过细胞外信号调节激酶(ERK)-NLRP3/半胱天冬酶 1 信号通路,并增加 hNEC 中白细胞介素 1β和白细胞介素 18 的分泌。此外,IL-17A 诱导的细胞焦亡通过影响糖皮质激素受体-α和糖皮质激素受体-β的表达导致类固醇耐药,抑制细胞焦亡蛋白部分消除了 hNEC 中 IL-17A 诱导的类固醇耐药。
在 CRSwNP 患者中,升高的 IL-17A 水平通过 ERK-NLRP3/半胱天冬酶 1 信号通路促进 hNEC 细胞焦亡,并通过影响糖皮质激素受体的动态平衡导致糖皮质激素耐药。