Increased inflammatory mediators in the ocular surface tissue in keratoconus.

IF 1.8 3区医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Vision Pub Date : 2024-09-02 eCollection Date: 2024-01-01

Albert Santos, José A P M Filho, Marcos A Cenedeze, Meire I Hiyane, Mariane T Amano, Mario C Cruz, Flavio E Hirai, Niels O S Camara, Luciene B de Sousa, Lauro A de Oliveira

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Abstract

Purpose: This study aimed to characterize the inflammatory mediators present in the tear film of patients with keratoconus (KC). It also aimed to investigate the gene expression of these mediators in corneal epithelial cells and their immune activity in conjunctival epithelial cells in patients with KC compared to a control group.

Methods: This transversal study included 30 patients with KC and 23 control group participants. Tear samples were collected by washing the ocular surface with 60 μL of sterile buffered saline solution. The levels of interleukin IL-5, IL-13, IL-2, IL-6, IL-10, interferon-gamma, tumor necrosis factor-alpha, and IL-4 were measured using a LEGEND plex HU Th1/Th2 panel kit and analyzed using flow cytometry. Corneal epithelial samples were obtained via manual keratectomy from KC patients scheduled for corneal crosslinking and from individuals scheduled for photorefractive keratectomy (control group). These samples were immediately stored at -70 °C for mRNA extraction and subsequent reverse transcription polymerase chain reaction analysis to measure IL-5 and IL-6 gene expression. Conjunctival epithelium samples were collected using impression cytology and analyzed using immunohistochemistry and confocal microscopy to detect IL-5 and IL-6 immunoreactions.

Results: Our study found no statistically significant differences in the tear film cytokine concentrations between the two groups. In addition, the gene expression of IL-5 and IL-6 in the corneal epithelium was higher in the KC group than in the control group, with IL-5 showing a 50% increase and IL-6 showing a 20% increase. Immunohistochemical analysis revealed a greater immunostaining of IL-5 and IL-6 in the conjunctival epithelium of patients with KC compared to the control group.

Conclusions: In this study, despite higher levels of IL-5 and IL-6 in the tear film of patients with KC, there was no statistically significant difference compared to the control group. However, there was heightened immune activity in the corneal and conjunctival epithelial cells of patients with KC based on IL-5 and IL-6 gene expression and their immunodetection, respectively.

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角膜炎患者眼表组织中的炎症介质增多。

目的：本研究旨在确定角膜炎（KC）患者泪膜中炎症介质的特征。与对照组相比，本研究还旨在调查 KC 患者角膜上皮细胞中这些介质的基因表达及其在结膜上皮细胞中的免疫活性：这项横向研究包括 30 名 KC 患者和 23 名对照组参与者。用 60 μL 无菌缓冲生理盐水冲洗眼表后收集泪液样本。使用 LEGEND plex HU Th1/Th2 面板试剂盒测量白细胞介素 IL-5、IL-13、IL-2、IL-6、IL-10、γ 干扰素、肿瘤坏死因子-α 和 IL-4 的水平，并使用流式细胞术进行分析。通过人工角膜切除术从计划接受角膜交联术的 KC 患者和计划接受光屈光性角膜切除术的患者（对照组）处获取角膜上皮样本。这些样本被立即保存在零下 70 摄氏度的环境中，用于提取 mRNA 和随后的反转录聚合酶链反应分析，以测量 IL-5 和 IL-6 基因的表达。使用印模细胞学方法收集结膜上皮样本，并使用免疫组化和共聚焦显微镜分析以检测 IL-5 和 IL-6 的免疫反应：结果：我们的研究发现，两组患者的泪膜细胞因子浓度在统计学上没有显著差异。此外，KC 组角膜上皮细胞中 IL-5 和 IL-6 的基因表达量高于对照组，其中 IL-5 增加了 50%，IL-6 增加了 20%。免疫组化分析显示，与对照组相比，KC 患者结膜上皮中的 IL-5 和 IL-6 免疫染色更高：在这项研究中，尽管 KC 患者泪膜中的 IL-5 和 IL-6 水平较高，但与对照组相比并无统计学意义上的显著差异。然而，根据 IL-5 和 IL-6 基因表达及其免疫检测结果，KC 患者角膜和结膜上皮细胞的免疫活性增强。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Molecular Vision 生物-生化与分子生物学

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

1 months

期刊介绍： Molecular Vision is a peer-reviewed journal dedicated to the dissemination of research results in molecular biology, cell biology, and the genetics of the visual system (ocular and cortical). Molecular Vision publishes articles presenting original research that has not previously been published and comprehensive articles reviewing the current status of a particular field or topic. Submissions to Molecular Vision are subjected to rigorous peer review. Molecular Vision does NOT publish preprints. For authors, Molecular Vision provides a rapid means of communicating important results. Access to Molecular Vision is free and unrestricted, allowing the widest possible audience for your article. Digital publishing allows you to use color images freely (and without fees). Additionally, you may publish animations, sounds, or other supplementary information that clarifies or supports your article. Each of the authors of an article may also list an electronic mail address (which will be updated upon request) to give interested readers easy access to authors.