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Updated: Jun 27, 2025
Establishment of A Mouse Model of Aqueous Deficiency Dry Eye
Published on: November 1, 2024
Role of NADPH Oxidase 4 on Dry Eye Syndrome in Mice
Mian Guo1,2,3,4, Taixiang Liu1,2,3,4, Yuan Miao1,2,3,4
1Department of Ophthalmology, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou Province, China.
Abstract:
Objective: This study aims to investigate the effect of NADPH oxidase 4 (NOX4)-mediated inflammation on concanavalin A (ConA)-induced dry eye syndrome (DES) in mice. Methods: Thirty-six mice were randomly divided into Control, Model, no-load Control, and NOX4 interference group. Adenovirus was injected (10 μL) into the lacrimal glands of both eyes of mice in no-load Control group and NOX4 interference group. Four days after adenovirus injection, the Control group was injected with phosphate-buffered saline, and the other groups were injected with ConA (200 μg) in the lacrimal glands of mice to establish DES models. The tear secretion rate was estimated by phenol red thread test. Lissamine green eye staining was used to evaluate conjunctival damage. The corneal surface was observed by hematoxylin-eosin (HE) staining and scanning electron microscopy (SEM). The morphology and quantity of conjunctival epithelial cells and goblet cells were observed by Periodic acid-Schiff staining. The expression of NOX4, NOD-like receptor thermal protein domain-associated protein 3 (NLRP3), interleukin-1β (IL-1β), and mucin 5 subtype AC (MUC5AC) was detected by immunohistochemistry. Results: Compared with the Control group, the Model group showed a significant decrease in tear secretion and an upregulation in microscopic image score. The HE staining and SEM showed corneal and conjunctiva damage in the Model group. The protein expression of NOX4, NLRP3, and IL-1β was upregulated, but MUC5AC was downregulated in the Model group. After interfering with NOX4, all these indicators were reversed. Conclusion: The pathological process of concanavalin A-induced DES appears to be related to NOX4.
Insights
Concanavalin A-induced dry eye syndrome (DES) in mice involves NADPH oxidase 4 (NOX4)-mediated inflammation. Inhibiting NOX4 reversed key indicators, suggesting NOX4
Area of Science:
- Ophthalmology
- Immunology
- Molecular Biology
Background:
- Dry eye syndrome (DES) is a prevalent ocular surface disease characterized by inflammation and damage.
- NADPH oxidase 4 (NOX4) is implicated in inflammatory processes, but its specific role in DES remains unclear.
Purpose of the Study:
- To investigate the role of NOX4-mediated inflammation in concanavalin A (ConA)-induced dry eye syndrome (DES) in a mouse model.
- To determine if interfering with NOX4 can ameliorate the pathological features of DES.
Main Methods:
- Concanavalin A (ConA) was used to induce DES in mice, with a NOX4 interference group established using adenovirus.
- Tear secretion, conjunctival damage, corneal surface morphology, and goblet cell counts were assessed.
- The expression of NOX4, NLRP3 inflammasome components (NLRP3, IL-1β), and MUC5AC was analyzed via immunohistochemistry.
Main Results:
- ConA-induced DES mice exhibited reduced tear secretion, increased conjunctival/corneal damage, and elevated NOX4, NLRP3, and IL-1β expression.
- MUC5AC expression was decreased in the DES model.
- Interference with NOX4 led to a reversal of these pathological changes, improving tear secretion and reducing inflammation.
Conclusions:
- NOX4-mediated inflammation plays a significant role in the pathogenesis of ConA-induced dry eye syndrome.
- Targeting NOX4 presents a potential therapeutic strategy for managing dry eye disease.
