Li S X, Li X, Chen X H, Wu X F, Huang J C, Lai X P, Yang L Y, Huang Z Z, Zhang G H, Chang L H
Department of Otorhinolaryngology Head and Neck Surgery, the Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510630, China.
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2021 Feb 3;56(2):124-132. doi: 10.3760/cma.j.cn115330-20200814-00675.
To investigate the correlation between Notch pathway expression in nasal polyps and Treg percentage and Eos infiltration. Patients with chronic sinusitis and simple nasal septum deviation who received nasal endoscopic surgery in the Third Affiliated Hospital of Sun Yat-Sen University between November 2012 and August 2018 were selected and enrolled in CRS group and control group respectively. Nasal mucosa tissues were collected from 30 CRSsNP patients (14 males and 16 females aged from 18 to 63), 58 CRSwNP patients (38 males and 20 females aged from 18 to 65) and 29 patients (19 males and 10 females aged from 20 to 57), who underwent nasal endoscopic surgery for correction of simple nasal septum deviation. Hematoxylin-eosin(HE) staining was used to observe the infiltration of eosinophilic granulocytes in the tissues and to classify chronic sinusitis with polyps (CRSwNP) into eosinophilic chronic rhinosinusitis with nasal polyps (Eos-CRSwNP)and non-eosinophilic chronic rhinosinusitis with nasal polyps (Eos-CRSwNP). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of Notch pathway receptors (Notch-l, 2, 3, 4) and their ligands (Jagded-l, Jagded-2, Delta-l, Delta-3and Delta-4) in the nasal mucosa of each group, as well as the expression of Th2 cytokines (IL-4, IL-5, IL-13), eosinophilic cationic protein (ECP)and the key transcription factor Foxp3 in Treg cells. Finally, flow cytometry was used to detect CD4CD25Foxp3 Treg cells in nasal mucosa of each group. Compared with controls, the expression of Th2 cytokines (IL-4, IL-5, IL-13) in CRSsNP and non-Eos-CRSwNP patients was the highest in Eos-CRSwNP (=16.930,9.197,9.116, all <0.05). Foxp3 had the lowest expression in Eos-CRSwNP patients and was lower than non-Eos-CRSwNP patients (=2.780,<0.05), and was negatively correlated with ECP (=-0.326,<0.05). Compared with controls, Eos-CRSwNP patients in CRSsNP patients and non-Eos-CRSwNP patients exhibited a significantly lower frequency of CD4CD25Foxp3Treg cells (=13.140, all <0.01). The expression of Notch-l and Jagged-l in Eos-CRSwNP was significantly higher than that of the controls, CRSsNP patients and non-Eos-CRSwNP patients (=5.953/=6.380, <0.05). In the nasal polyp group, the expression of Notch-l and Jagged-l showed significantly negative correlation with Foxp3 (=-0.611/-0.346, all <0.05), and positive correlation with Th2 cytokines (IL-4, IL-5, IL-13) and ECP, respectively (=0.781/0.459,0.621/0.601,0.605/0.490,0.464/0.668, all <0.05). There was no significant difference in the expression of receptor and ligand of the other Notch pathway among the groups. Abnormal activation of Notch-l/Jagged-l pathway may be involved in decreasing Treg ratio in Eos-CRSwNP, thereby promoting Th2 inflammatory response and Eosinophil infiltration.
探讨鼻息肉中Notch信号通路表达与调节性T细胞(Treg)百分比及嗜酸性粒细胞浸润之间的相关性。选取2012年11月至2018年8月在中山大学附属第三医院接受鼻内镜手术的慢性鼻窦炎合并单纯鼻中隔偏曲患者,分别纳入慢性鼻窦炎伴鼻息肉(CRSwNP)组和对照组。收集30例CRSsNP患者(男14例,女16例,年龄18~63岁)、58例CRSwNP患者(男38例,女20例,年龄18~65岁)及29例因单纯鼻中隔偏曲行鼻内镜手术矫正的患者(男19例,女10例,年龄20~57岁)的鼻黏膜组织。采用苏木精-伊红(HE)染色观察组织中嗜酸性粒细胞浸润情况,并将鼻息肉型慢性鼻窦炎(CRSwNP)分为嗜酸性粒细胞性鼻息肉型慢性鼻窦炎(Eos-CRSwNP)和非嗜酸性粒细胞性鼻息肉型慢性鼻窦炎(non-Eos-CRSwNP)。采用实时定量聚合酶链反应(qRT-PCR)检测各组鼻黏膜中Notch信号通路受体(Notch-1、2、3、4)及其配体(Jagged-1、Jagged-2、Delta-1、Delta-3和Delta-4)的表达,以及Th2细胞因子(IL-4、IL-5、IL-13)、嗜酸性粒细胞阳离子蛋白(ECP)和Treg细胞中关键转录因子Foxp3的表达。最后,采用流式细胞术检测各组鼻黏膜中CD4+CD25+Foxp3+Treg细胞。与对照组相比,CRSsNP和non-Eos-CRSwNP患者中Th2细胞因子(IL-4、IL-5、IL-13)在Eos-CRSwNP中的表达最高(F=16.930、9.197、9.116,均P<0.05)。Foxp3在Eos-CRSwNP患者中的表达最低,且低于non-Eos-CRSwNP患者(t=2.780,P<0.05),且与ECP呈负相关(r=-0.326,P<0.05)。与对照组相比,CRSsNP患者和non-Eos-CRSwNP患者中的Eos-CRSwNP患者CD4+CD25+Foxp3+Treg细胞频率显著降低(F=13.140,均P<0.01)。Eos-CRSwNP中Notch-1和Jagged-1的表达显著高于对照组、CRSsNP患者和non-Eos-CRSwNP患者(t=5.953/t=6.380,P<0.05)。在鼻息肉组中,Notch-1和Jagged-1的表达与Foxp3呈显著负相关(r=-0.611/r=-0.346,均P<0.05),分别与Th2细胞因子(IL-4、IL-5、IL-13)和ECP呈正相关(r=0.781/0.459、0.621/0.601、0.605/0.490、0.464/0.668,均P<0.05)。各组中其他Notch信号通路受体和配体的表达无显著差异。Notch-1/Jagged-1信号通路的异常激活可能参与降低Eos-CRSwNP中Treg比例,从而促进Th2炎症反应和嗜酸性粒细胞浸润。