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3-Methyladenine Alleviates Experimental Subretinal Fibrosis by Inhibiting Macrophages and M2 Polarization Through the
Qiyu Bo1, Mengxi Shen1, Meichun Xiao1
1Department of Ophthalmology, Shanghai General Hospital (Shanghai First People's Hospital), Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Abstract:
To explore the effects of 3-methyladenine (3-MA), a selective inhibitor of phosphatidylinositol-3-kinase (PI3K), on experimental subretinal fibrosis (SRF) in mice. The SRF mouse model was established by 532 nm laser photocoagulation at each fundus of mice on day 0. 3-MA was administered every 2 days from day 0 to 35. Immunofluorescence of choroidal flat mounts was performed to evaluate the size of SRF area, local macrophages, and polarization, respectively. Besides, Western blot analysis was carried out to assess the expression levels of macrophage polarization-related genes, Arg-1, Ym-1, and transforming growth factor-β2 (TGF-β2). Co-culture and migration experiments were used to demonstrate the inhibitory effect of 3-MA on fibroblasts. The gene knockout and Western blot analysis were used to explore the signal pathways related to macrophage polarization. Compared with the control group, the 3-MA-treated group showed significantly less size of SRF area. 3-MA treatment reduced both circulating and local macrophages, and counteracted M2 polarization. Moreover, 3-MA inhibited fibroblast recruitment. Mechanistically, we proved that 3-MA inhibits macrophage M2 polarization by suppressing PI3K/Akt signal pathway rather than the PI3K-autophagy-related signal pathway. 3-MA exerts antifibrotic effects on experimental SRF by targeting circulating and local macrophages and M2 polarization, through PI3K/Akt signal pathway. These results support the potential use of 3-MA as a new therapeutic modality for SRF associated with neovascular age-related macular degeneration.
Insights
3-methyladenine (3-MA) reduces experimental subretinal fibrosis (SRF) by targeting macrophages and M2 polarization via the PI3K/Akt pathway, offering potential therapeutic benefits for related eye conditions.
Area of Science:
- Ophthalmology
- Immunology
- Pharmacology
Background:
- Subretinal fibrosis (SRF) is a significant complication of neovascular age-related macular degeneration.
- Current treatments for SRF are limited, necessitating novel therapeutic strategies.
Purpose of the Study:
- To investigate the antifibrotic effects of 3-methyladenine (3-MA), a phosphatidylinositol-3-kinase (PI3K) inhibitor, on experimental SRF in a mouse model.
- To elucidate the underlying mechanisms of 3-MA's action, focusing on macrophage polarization and signaling pathways.
Main Methods:
- An experimental SRF mouse model was induced using laser photocoagulation.
- 3-MA was administered systemically, and its effects on SRF area, macrophage infiltration and polarization, and fibroblast activity were assessed.
- Immunofluorescence, Western blot, co-culture, and gene knockout techniques were employed to analyze macrophage polarization and signaling pathways (PI3K/Akt).
Main Results:
- 3-MA treatment significantly reduced the size of the SRF area compared to controls.
- 3-MA decreased both circulating and local macrophage populations and inhibited M2 polarization.
- 3-MA suppressed fibroblast recruitment and migration, acting through the PI3K/Akt signaling pathway to modulate macrophage M2 polarization.
Conclusions:
- 3-MA demonstrates potent antifibrotic effects in experimental SRF by targeting macrophages and M2 polarization via the PI3K/Akt pathway.
- These findings suggest 3-MA as a potential therapeutic agent for SRF, particularly in the context of neovascular age-related macular degeneration.
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