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Updated: Sep 19, 2025
Author Spotlight: Decoding Corneal Neovascularization with Alkali Burn Model for Future Therapeutic Strategies
Published on: June 30, 2023
RIPK3-Mediated Necroptosis Drives Macrophage Infiltration and Corneal Neovascularization After Alkali Burn
1Department of Ophthalmology, The People's Hospital of Guangxi Zhuang Autonomous Region, Guangxi Key Laboratory of Eye Health, Guangxi Health Commission Key Laboratory of Ophthalmology and Related Systemic Diseases Artificial Intelligence Screening Technology, Institute of Ophthalmic Diseases, Guangxi Academy of Medical Sciences, Guangxi, China.
Purpose:
To reveal the role of receptor-interacting protein kinase 3 (RIPK3) in regulating macrophage inflammation and corneal neovascularization (CoNV) induced by alkali burn.
Methods:
A corneal alkali burn (AB) model was established in C57BL/6J (wild-type) and RIPK3fl/flCx3cr1+/cre (RIPK3-/-, RIPK3 knockout [KO]) mice using sodium hydroxide. Anterior segment optical coherence tomography and hematoxylin and eosin staining were used to evaluate the impact of RIPK3 on corneal edema and morphology. CoNV was detected by slit-lamp microscopy and whole-mount immunofluorescence staining of cornea. Corneal macrophage and necroptotic cell death was analyzed through immunofluorescence staining and propidium iodide (PI) staining. Activation of the necroptosis pathway was examined after corneal AB by western blot.
Results:
Necroptosis-related proteins RIPK1, RIPK3, and mixed lineage kinase domain-like (MLKL) were upregulated and activated following corneal AB. Among these, RIPK3 demonstrated the most pronounced increase. Notably, the elevated level of RIPK3 was prominently colocalized with the infiltrating F4/80+ macrophages. RIPK3 KO significantly alleviated corneal edema and morphology defects. Additionally, as the corneal morphological defects progressed, macrophages became activated, and CoNV and lymphangiogenesis (LyG) were enhanced. RIPK3 KO markedly reduced AB-induced macrophage accumulation, as well as CoNV and LyG. RIPK3 KO mice also showed a meaningful decrease in PI+ necroptotic cells. Mechanistically, AB-induced necroptosis stimulated the expression of MLKL and fibroblast growth factor 2 (FGF2), whereas RIPK3 deficiency decreased their expression.
Conclusions:
This study revealed that RIPK3-mediated necroptosis drives macrophage inflammation and CoNV. Targeting RIPK3 could effectively suppress these responses by inhibiting the MLKL/FGF2 pathway, making it a promising therapeutic strategy for corneal AB.
Insights
Receptor-interacting protein kinase 3 (RIPK3) drives inflammation and corneal neovascularization after alkali burns. Inhibiting RIPK3 suppresses these responses, offering a potential therapeutic strategy for corneal alkali burns.
Area of Science:
- Ophthalmology
- Immunology
- Cell Biology
Background:
- Corneal alkali burns (AB) trigger inflammation and neovascularization.
- The role of receptor-interacting protein kinase 3 (RIPK3) in AB-induced responses is not fully understood.
- Necroptosis is a regulated form of cell death implicated in inflammatory diseases.
Purpose of the Study:
- To investigate the role of RIPK3 in regulating macrophage inflammation and corneal neovascularization (CoNV) following alkali burn injury.
- To elucidate the molecular mechanisms by which RIPK3 influences these pathological processes.
Main Methods:
- Established a corneal alkali burn model in wild-type and RIPK3 knockout mice.
- Assessed corneal edema, morphology, neovascularization, and lymphangiogenesis using imaging and histological techniques.
- Analyzed macrophage infiltration, necroptotic cell death, and necroptosis pathway activation via immunofluorescence, PI staining, and western blot.
Main Results:
- Corneal alkali burns upregulated and activated necroptosis pathway proteins, with RIPK3 showing the most significant increase and colocalizing with macrophages.
- RIPK3 knockout significantly reduced corneal edema, improved morphology, and decreased macrophage accumulation.
- RIPK3 deficiency markedly reduced AB-induced corneal neovascularization (CoNV) and lymphangiogenesis (LyG) by inhibiting MLKL and FGF2 expression.
Conclusions:
- RIPK3-mediated necroptosis is a key driver of macrophage inflammation and CoNV in corneal alkali burns.
- Targeting RIPK3 and its downstream MLKL/FGF2 pathway presents a promising therapeutic strategy for managing corneal alkali burns.
