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AZD6738 Attenuates LPS-Induced Corneal Inflammation and Fibrosis by Modulating Macrophage Function and Polarization
Longxiang Huang1,2,3,4, Youfang Luo5
1Department of Ophthalmology, The First Affiliated Hospital of Fujian Medical University, Fuzhou, China. huanglx626@163.com.
Abstract:
This study aimed to evaluate the therapeutic potential of AZD6738, an ATR inhibitor, in LPS-induced bacterial keratitis (BK) by targeting macrophage function and polarization. A murine model of LPS-induced BK was established, with AZD6738 (100 µM) administered subconjunctivally and topically. Corneal opacity, edema, and inflammation were assessed using slit-lamp microscopy and histological analysis. Macrophage infiltration and fibrosis were evaluated via immunofluorescence, qPCR, and Western blotting. In vitro, RAW264.7 cells were treated with 2.5 µM AZD6738 to examine its effects on cell viability, oxidative stress, and inflammation-related gene expression. AZD6738 significantly reduced corneal opacity, thickness, and neovascularization in LPS-treated mice. It suppressed macrophage infiltration, collagen deposition, and pro-inflammatory cytokine expression. In RAW264.7 cells, AZD6738 induced cell death, elevated ROS production, and downregulated inflammatory markers. ATR inhibition mitigated NF-κB activation and modulated macrophage polarization, attenuating M1 pro-inflammatory responses. AZD6738 effectively alleviates LPS-induced corneal inflammation and fibrosis by regulating macrophage function and polarization via the NF-κB signaling pathway. ATR inhibition represents a promising therapeutic strategy for the treatment of corneal inflammation.
Insights
AZD6738, an ATR inhibitor, effectively treats bacterial keratitis by reducing corneal inflammation and fibrosis. It targets macrophage function and polarization, offering a promising therapeutic strategy for eye inflammation.
Area of Science:
- Ophthalmology
- Immunology
- Pharmacology
Background:
- Bacterial keratitis (BK) is an inflammatory eye condition often leading to vision loss.
- Macrophage dysfunction and polarization play critical roles in BK pathogenesis.
- Current treatments for BK may have limitations, necessitating novel therapeutic approaches.
Purpose of the Study:
- To evaluate the therapeutic potential of AZD6738, a novel ATR inhibitor, in a murine model of LPS-induced bacterial keratitis.
- To investigate the effects of AZD6738 on macrophage function, polarization, and inflammatory pathways in BK.
- To assess the impact of AZD6738 on corneal opacity, edema, inflammation, and fibrosis.
Main Methods:
- A murine model of LPS-induced bacterial keratitis was established.
- AZD6738 was administered subconjunctivally and topically.
- Corneal opacity, edema, and inflammation were assessed using slit-lamp microscopy and histology.
- Macrophage infiltration, fibrosis, and inflammatory markers were evaluated using immunofluorescence, qPCR, and Western blotting.
- In vitro studies on RAW264.7 cells examined AZD6738's effects on cell viability, oxidative stress, and gene expression.
Main Results:
- AZD6738 significantly reduced corneal opacity, thickness, and neovascularization in LPS-treated mice.
- It suppressed macrophage infiltration, collagen deposition, and pro-inflammatory cytokine expression.
- In vitro, AZD6738 induced cell death, elevated reactive oxygen species (ROS) production, and downregulated inflammatory markers.
- ATR inhibition mitigated NF-κB activation and modulated macrophage polarization, attenuating M1 pro-inflammatory responses.
Conclusions:
- AZD6738 effectively alleviates LPS-induced corneal inflammation and fibrosis.
- The therapeutic effects are mediated by regulating macrophage function and polarization via the NF-κB signaling pathway.
- ATR inhibition represents a promising therapeutic strategy for treating corneal inflammation and related fibrotic conditions.
