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Title: [IL-25-regulated type 2 innate lymphoid cells activation promote allergic fungal rhinosinusitis].
Author: Zhou MH, Zhang SJ, Wang F, Zhao YL, Liu YQ, Zhang YJ, Liu S.
Journal: Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi; 2019 Jan 07; 54(1):38-45. PubMed ID: 30704168.
Abstract:
Objective: To investigate the regulation of IL-25 on type Ⅱ innate lymphoid cells (ILC2s) activation in the pathogenesis of allergic fungal rhinosinusitis (AFRS). Methods: Nasal mucosa tissues were collected from 16 AFRS patients and 12 patients, who underwent nasal endoscopic surgery for cerebrospinal rhinorrhea or skull base benign tumor during the period from June 2016 to June 2017 in Department of Rhinology, the First Affiliated Hospital of Zhengzhou University. Firstly, flow cytometry was used to detect ILC2s in nasal mucosa of both groups. Secondly, the expression of IL-25, IL-5 and IL-13 in nasal mucosa was detected by immunofluorescence and/or Western Blot assay. Finally, fungal extracts, IL-25 and glucocorticoids were used to stimulate nasal mucosal epithelial cells and tissues in vitro respectively to detect the regulatory effect of IL-25 on ILC2s. SPSS 16.0 software was used to analyze the data. Results: The prevalence of ILC2s in nasal tissues was higher in patients with AFRS than those of the control group ((3.85±1.52)%(Mean±SD) vs (0.32±0.10)%, U=9.00, P<0.05). There was a positive correlation between the prevalence of ILC2s and the number of eosinophils in nasal mucosa of patients with ARFS (r=0.80, P<0.05). The expression of IL-25, IL-5 and IL-13 in nasal mucosa epithelium of AFRS group was significantly higher than that of the control group (0.49±0.13 vs 0.23±0.09, 0.23±0.05 vs 0.10±0.04, 0.31±0.08 vs 0.14±0.07, t value was 5.90, 7.21, 5.69, respectively, all P<0.05). Fungal stimulation enhanced the expression of IL-25 protein in nasal epithelial cells of both groups (0.67±0.19 vs 0.25±0.12 (AFRS group), 0.62±0.17 vs 0.27±0.16 (control group), q value was 8.65, 9.26, respectively, all P<0.05). In the IL-25 stimulated nasal mucosa at a concentration of 1, 10 and 100 ng/ml, the expression level of retinoid acid-related orphan receptor α (RORα) mRNA was 2.07±1.53, 5.06±0.93, 7.38±2.30, respectively; the expression level of GATA binding protein 3 (GATA3) mRNA was 3.58±1.29, 6.14±1.55, 7.64±2.28, respectively; the expression level of IL-5 protein was 0.21±0.06, 0.32±0.06, 0.38±0.10, respectively; the expression level of IL-13 was 0.52±0.13, 0.69±0.22, 0.82±0.21, respectively, which were significantly higher than that in the unstimulated nasal mucosa (1.00±0.00, 1.00±0.00, 0.11±0.05, 0.35±0.15, F value was 63.45, 59.27, 49.35, 20.20, respectively, all P<0.05). The up-regulation could be inhibited by dexamethasone (F value was 89.20, 92.47, 99.63, 49.82, respectively, all P<0.05). Conclusions: Epithelial-derived IL-25 up-regulates the expression of IRC2s-dependent transcription factors RORα and GATA3 mRNA, which are positively correlated with elevated IL-13 and IL-5 expression levels in tissues, may be involved in AFRS inflammatory response, and are inhibited by glucocorticoids. 目的： 探讨白细胞介素（IL）25调控Ⅱ型固有淋巴细胞（type Ⅱ innate lymphoid cells，ILC2s）活化在变应性真菌性鼻窦炎（allergic fungal rhinosinusitis，AFRS）发病机制中的作用。 方法： 收集2016年6月至2017年6月期间，于郑州大学第一附属医院鼻科诊治的16例AFRS患者的鼻黏膜组织，并选取同期行鼻内镜脑脊液鼻漏修补术或颅底良性肿瘤切除术的12例患者的鼻黏膜组织作为对照组。首先应用流式细胞术检测两组鼻黏膜组织中的ILC2s，然后应用免疫荧光和（或）Western Blot的方法检测IL-25、IL-5和IL-13在鼻黏膜组织中的表达，最后应用真菌提取物、IL-25和糖皮质激素分别刺激体外培养的鼻黏膜上皮细胞和组织，检测IL-25对ILC2s的调控作用。以SPSS 16.0软件对数据进行统计学分析。 结果： AFRS组鼻黏膜组织中ILC2s百分比为（3.85±1.52）%（x±s，下同），明显高于对照组（0.32±0.10）%，二者差异具有统计学意义（U=9.00，P<0.05）。ARFS患者鼻黏膜组织中ILC2s百分比和嗜酸粒细胞数量呈正相关（r=0.80，P<0.05）。两组患者鼻黏膜上皮组织均有IL-25、IL-5和IL-13的表达，AFRS组的表达强度明显高于对照组（0.49±0.13比0.23±0.09，0.23±0.05比0.10±0.04，0.31±0.08比0.14±0.07，t值分别为5.90、7.21、5.69，P值均<0.05）。真菌刺激体外原代培养的鼻黏膜上皮细胞后，两组患者鼻黏膜上皮细胞的IL-25蛋白表达强度均升高（AFRS组为0.67±0.19比0.25±0.12，对照组为0.62±0.17比0.27±0.16，q值分别为8.65、9.26，P值均<0.05）。以浓度为1、10和100 ng/ml的IL-25刺激鼻黏膜组织12 h后，维甲酸相关孤儿核受体α（retinoid acid-related orphan receptor α，RORα）mRNA的表达量分别为2.07±1.53、5.06±0.93、7.38±2.30，GATA结合蛋白3（GATA binding protein 3，GATA3）mRNA的表达量分别为3.58±1.29、6.14±1.55、7.64±2.28，IL-5蛋白的表达量分别为0.21±0.06、0.32±0.06、0.38±0.10，IL-13蛋白的表达量分别为0.52±0.13、0.69±0.22、0.82±0.21，均高于刺激前（1.00±0.00、1.00±0.00、0.11±0.05、0.35±0.15），差异具有统计学意义（F值分别为63.45、59.27、49.35、20.20，P值均<0.05），地塞米松可抑制四者表达的上调（F值分别为89.20、92.47、99.63、49.82，P值均<0.05）。 结论： 上皮细胞衍生的IL-25上调了ILC2s依赖的转录因子RORα和GATA3 mRNA的表达，二者与组织中IL-13和IL-5表达水平升高呈正相关，可能参与了AFRS的炎性反应，并可受糖皮质激素抑制。.

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