Department of Special Medicine, School of Basic Medicine, Qingdao University, Qingdao, China.
Department of Anthropotomy and Histo-Embryology, School of Basic Medicine, Qingdao University, Qingdao, China.
Invest Ophthalmol Vis Sci. 2022 Jun 1;63(6):1. doi: 10.1167/iovs.63.6.1.
To investigate the expression of corneal epithelium-derived netrin-1 (NTN-1) and its immunoregulatory function in dry eye disease (DED) using a DED mouse model.
We generated DED mouse models with desiccating stress under scopolamine treatment. RNA sequencing was performed to identify differentially expressed genes (DEGs) in the corneal epithelium of DED mice. NTN-1 expression was analyzed via real-time PCR, immunofluorescence staining, and immunoblotting. The DED mice were then treated with recombinant NTN-1 or neutralizing antibodies to investigate the severity of the disease, dendritic cell (DC) activation, and inflammatory cytokine expression.
A total of 347 DEGs (292 upregulated and 55 downregulated) were identified in the corneal epithelium of DED mice: corneal epithelium-derived NTN-1 expression was significantly decreased in DED mice compared to that in control mice. Topical recombinant NTN-1 application alleviated the severity of the disease, accompanied by restoration of tear secretion and goblet cell density. In addition, NTN-1 decreased the number of DCs, inhibited the activation of the DCs and Th17 cells, and reduced the expression of inflammatory factors in DED mice. In contrast, blocking endogenous NTN-1 activity with an anti-NTN-1 antibody aggravated the disease, enhanced DC activation, and upregulated the inflammatory factors in the conjunctivae of DED mice.
We identified decreased NTN-1 expression in the corneal epithelium of DED mice. Our findings elucidate the role of NTN-1 in alleviating DED and impeding DC activation, thereby indicating its therapeutic potential in suppressing ocular inflammation in DED.
通过干燥性眼病（DED）小鼠模型，研究角膜上皮细胞衍生的神经导向因子-1（NTN-1）的表达及其在 DED 中的免疫调节功能。
我们通过东莨菪碱处理下的干燥应激，生成 DED 小鼠模型。通过 RNA 测序鉴定 DED 小鼠角膜上皮细胞中的差异表达基因（DEGs）。通过实时 PCR、免疫荧光染色和免疫印迹分析 NTN-1 的表达。然后用重组 NTN-1 或中和抗体处理 DED 小鼠，以研究疾病的严重程度、树突状细胞（DC）激活和炎症细胞因子的表达。
在 DED 小鼠的角膜上皮中鉴定出 347 个 DEGs（292 个上调，55 个下调）：与对照小鼠相比，DED 小鼠的角膜上皮源性 NTN-1 表达明显降低。局部应用重组 NTN-1 可减轻疾病严重程度，同时恢复泪液分泌和杯状细胞密度。此外，NTN-1 减少了 DC 的数量，抑制了 DC 和 Th17 细胞的激活，并降低了 DED 小鼠中炎症因子的表达。相反，用抗 NTN-1 抗体阻断内源性 NTN-1 活性会加重疾病，增强 DC 激活，并上调 DED 小鼠结膜中的炎症因子。
我们发现 DED 小鼠角膜上皮细胞中 NTN-1 表达降低。我们的研究结果阐明了 NTN-1 在缓解 DED 和抑制 DC 激活中的作用，表明其在抑制 DED 中的眼部炎症方面具有治疗潜力。
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