Intrinsic regeneration mechanisms of murine lacrimal gland epithelial cells for therapeutic applications in aqueous-deficient dry eye disease

IF 6.6 1区医学 Q1 OPHTHALMOLOGY

Ocular Surface Pub Date : 2026-07-01 Epub Date: 2026-06-13 DOI:10.1016/j.jtos.2026.06.007

Ingrid Zahn, Hannah Franke, Sophie Gleixner, Nicolas Blavet, Jana Dietrich, Philipp Arnold, Friedrich Paulsen

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引用次数: 0

Abstract

Purpose

Aqueous-deficient dry eye disease (ADDE) results from lacrimal gland dysfunction leading to tear film instability and chronic ocular surface inflammation. Despite the gland's known regenerative potential, the intrinsic molecular mechanisms enabling epithelial recovery remain poorly understood. This study aimed to characterize intrinsic regeneration processes of murine lacrimal gland epithelial cells (mLGECs) and identify pathways with therapeutic potential.

Methods

Primary mLGECs were isolated from neonatal C57BL/6J mice and characterized. Acute injury was induced by brief exposure to 20% ethanol. Metabolic activity, apoptosis, and transcriptional responses were analyzed during early recovery phases. Pathway enrichment was conducted using Gene Ontology and KEGG annotations. Regeneration-associated growth factors were validated by qPCR, ELISA, and metabolic activity and wound closure assays in the presence or absence of the MAPK inhibitor SCH772984. MAPK activation was assessed by ERK1/2 phosphorylation.

Results

mLGECs retained expression of epithelial and lacrimal-specific markers and exhibited regulated secretory activity. RNA sequencing and ERK1/2 phosphorylation revealed activation of MAPK signaling, accompanied by strong upregulation of regenerative ligands including Hbegf, Areg, Ngf, and Gdnf. Transient protein induction of HBEGF and AREG was confirmed. Functionally, HBEGF, AREG, NGF, and GDNF markedly enhanced metabolic recovery and wound closure, whereas pharmacological MAPK inhibition markedly impaired recovery.

Conclusion

These results identify MAPK signaling as a major component of the intrinsic epithelial injury response in lacrimal gland cells. HBEGF and AREG emerged as prominent mediators associated with enhanced recovery. The findings provide a mechanistic basis for developing defined growth factor–based approaches to support lacrimal gland epithelial regeneration.

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小鼠泪腺上皮细胞内在再生机制在缺水型干眼病治疗中的应用

目的：缺水性干眼病（ADDE）是由泪腺功能障碍导致泪膜不稳定和慢性眼表炎症引起的。尽管已知腺体具有再生潜能，但使上皮细胞恢复的内在分子机制仍然知之甚少。本研究旨在表征小鼠泪腺上皮细胞（mLGECs）的内在再生过程，并确定具有治疗潜力的途径。方法：从新生小鼠C57BL/6J中分离原发mlgec并进行特征鉴定。短暂暴露于20%乙醇诱导急性损伤。在早期恢复阶段分析代谢活性、细胞凋亡和转录反应。利用Gene Ontology和KEGG注释进行途径富集。在MAPK抑制剂SCH772984存在或不存在的情况下，通过qPCR、ELISA、代谢活性和伤口闭合试验验证再生相关生长因子。通过ERK1/2磷酸化来评估MAPK的激活。结果：mlges保留了上皮和泪腺特异性标志物的表达，并表现出调节的分泌活性。RNA测序和ERK1/2磷酸化显示MAPK信号被激活，并伴有再生配体（包括Hbegf、Areg、Ngf和Gdnf）的强烈上调。证实了HBEGF和AREG的瞬时蛋白诱导作用。在功能上，HBEGF、AREG、NGF和GDNF显著促进代谢恢复和伤口愈合，而药理MAPK抑制显著损害恢复。结论：这些结果表明MAPK信号是泪腺细胞内禀上皮损伤反应的主要组成部分。HBEGF和AREG是与增强恢复相关的重要介质。研究结果为开发明确的生长因子为基础的方法来支持泪腺上皮再生提供了机制基础。

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

Ocular Surface 医学-眼科学

CiteScore

11.60

自引率

14.10%

发文量

审稿时长

39 days

期刊介绍： The Ocular Surface, a quarterly, a peer-reviewed journal, is an authoritative resource that integrates and interprets major findings in diverse fields related to the ocular surface, including ophthalmology, optometry, genetics, molecular biology, pharmacology, immunology, infectious disease, and epidemiology. Its critical review articles cover the most current knowledge on medical and surgical management of ocular surface pathology, new understandings of ocular surface physiology, the meaning of recent discoveries on how the ocular surface responds to injury and disease, and updates on drug and device development. The journal also publishes select original research reports and articles describing cutting-edge techniques and technology in the field. Benefits to authors We also provide many author benefits, such as free PDFs, a liberal copyright policy, special discounts on Elsevier publications and much more. Please click here for more information on our author services. Please see our Guide for Authors for information on article submission. If you require any further information or help, please visit our Support Center