Bequignon Emilie, Mangin David, Bécaud Justine, Pasquier Jennifer, Angely Christelle, Bottier Mathieu, Escudier Estelle, Isabey Daniel, Filoche Marcel, Louis Bruno, Papon Jean-François, Coste André
Service d'Oto-Rhino-Laryngologie et de Chirurgie cervico-faciale, AP-HP, Hôpital Henri Mondor et Centre Hospitalier Intercommunal de Créteil, 94010, Créteil, France.
INSERM, U955, Equipe 13, Faculte de Medecine, 8 rue du General Sarrail, 94010, Créteil, France.
J Transl Med. 2020 Mar 24;18(1):136. doi: 10.1186/s12967-020-02309-9.
Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by an alteration in airway epithelial cell functions including barrier function, wound repair mechanisms, mucociliary clearance. The mechanisms leading to epithelial cell dysfunction in nasal polyps (NPs) remain poorly understood. Our hypothesis was that among the inflammatory cytokines involved in NPs, IL-6 could alter epithelial repair mechanisms and mucociliary clearance. The aim of this study was to evaluate the in vitro effects of IL-6 on epithelial repair mechanisms in a wound repair model and on ciliary beating in primary cultures of Human Nasal Epithelial Cells (HNEC).
Primary cultures of HNEC taken from 38 patients during surgical procedures for CRSwNP were used in an in vitro model of wound healing. Effects of increasing concentrations of IL-6 (1 ng/mL, 10 ng/mL, and 100 ng/mL) and other ILs (IL-5, IL-9, IL-10) on wound closure kinetics were compared to cultures without IL-modulation. After wound closure, the differentiation process was characterized under basal conditions and after IL supplementation using cytokeratin-14, MUC5AC, and β tubulin as immunomarkers of basal, mucus, and ciliated cells, respectively. The ciliated edges of primary cultures were analyzed on IL-6 modulation by digital high-speed video-microscopy to measure: ciliary beating frequency (CBF), ciliary length, relative ciliary density, metachronal wavelength and the ciliary beating efficiency index.
Our results showed that: (i) IL-6 accelerated airway wound repair in vitro, with a dose-response effect whereas no effect was observed after other ILs-stimulation. After 24 h, 79% of wounded wells with IL6-100 were fully repaired, vs 46% in the IL6-10 group, 28% in the IL6-1 group and 15% in the control group; (ii) specific migration analyses of closed wound at late repair stage (Day 12) showed IL-6 had the highest migration compared with other ILs (iii) The study of the IL-6 effect on ciliary function showed that CBF and metachronal wave increased but without significant modifications of ciliary density, length of cilia and efficiency index.
The up-regulated epithelial cell proliferation observed in polyps could be induced by IL-6 in the case of prior epithelial damage. IL-6 could be a major cytokine in NP physiopathology.
伴鼻息肉的慢性鼻-鼻窦炎（CRSwNP）的特征是气道上皮细胞功能改变，包括屏障功能、伤口修复机制、黏液纤毛清除功能。导致鼻息肉（NP）上皮细胞功能障碍的机制仍知之甚少。我们的假设是，在参与NP的炎性细胞因子中，白细胞介素-6（IL-6）可改变上皮修复机制和黏液纤毛清除功能。本研究的目的是评估IL-6在伤口修复模型中对上皮修复机制以及在人鼻上皮细胞（HNEC）原代培养物中对纤毛摆动的体外作用。
从38例接受CRSwNP手术的患者身上获取的HNEC原代培养物用于伤口愈合的体外模型。将浓度递增的IL-6（1 ng/mL、10 ng/mL和100 ng/mL）及其他白细胞介素（IL-5、IL-9、IL-10）对伤口闭合动力学的影响与未进行IL调节的培养物进行比较。伤口闭合后，在基础条件下以及补充IL后，分别使用细胞角蛋白-14、MUC5AC和β微管蛋白作为基底细胞、黏液细胞和纤毛细胞的免疫标志物来表征分化过程。通过数字高速视频显微镜分析IL-6调节下原代培养物的纤毛边缘，以测量：纤毛摆动频率（CBF）、纤毛长度、相对纤毛密度、同步波长和纤毛摆动效率指数。
我们的结果表明：（i）IL-6在体外加速气道伤口修复，呈剂量反应效应，而其他IL刺激后未观察到效果。24小时后，IL6-100处理的受伤孔中有79%完全修复，而IL6-10组为46%，IL6-1组为28%，对照组为15%；（ii）在修复后期（第12天）对闭合伤口的特异性迁移分析表明，与其他IL相比，IL-6的迁移率最高；（iii）对IL-6对纤毛功能影响的研究表明，CBF和同步波增加，但纤毛密度、纤毛长度和效率指数无显著变化。
在先前存在上皮损伤的情况下，息肉中观察到的上皮细胞增殖上调可能由IL-6诱导。IL-6可能是NP病理生理学中的主要细胞因子。
