The mechanism of serpinA3N alleviates retinal neural injury in diabetic mice by inhibiting Müller cell inflammation
10.13389/j.cnki.rao.2025.0163
- VernacularTitle:SerpinA3N通过抑制Müller细胞炎症减轻糖尿病小鼠视网膜神经损伤的作用机制研究
- Author:
Shilei SUN
1
;
Tao LÜ
;
Zhongfu ZUO
;
Chuang FENG
Author Information
1. 118000 辽宁省丹东市,丹东市第一医院眼科
- Publication Type:Journal Article
- Keywords:
diabetic retinopathy;
Müller cells;
SerpinA3N;
inflammatory response
- From:
Recent Advances in Ophthalmology
2025;45(12):949-955
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the mechanism of serine protease inhibitor A3N(SerpinA3N)in alleviating reti-nal neural injury in diabetic mice by inhibiting Müller cell inflammation.Methods Thirty-six db/db mice(72 eyes)were randomly divided into the db/db group,the db/db+SerpinA3N-overexpressing adeno-associated virus(AAV-SerpinA3N)group,and the db/db+empty vector adeno-associated virus(NC-SerpinA3N)group,with 12 mice in each group.Twelve age-matched healthy male littermate mice were randomly selected as the healthy control group(db/m group).Mice in each group were sacrificed 4 weeks after the corresponding treatments.Immunofluorescence staining was used to detect the co-localization of SerpinA3N and glial fibrillary acidic protein(GFAP)in the mouse retina.Hematoxylin-eosin(HE)staining was used to observe histopathological changes in the retinal tissue.TUNEL staining was used to detect the apoptosis of reti-nal ganglion cells(RGCs).Immunohistochemistry was used to detect GFAP expression.ELISA was used to measure the levels of inflammatory factors[interleukin(IL)-1 β,IL-6,IL-18,and tumor necrosis factor-α(TNF-α)]in the retinal tis-sue.The predicted target genes of SerpinA3N were imported into the STRING database to construct a protein-protein inter-action(PPI)network.The highest-scoring target was selected based on the scores for molecular docking.Western blot was used to detect the expression levels of SerpinA3N,nuclear factor kappa B(NF-κB),and spleen focus-forming virus proviral integration oncogene(Spi1)proteins in the retinal tissue.Results Immunofluorescence staining showed co-lo-calized expression of SerpinA3N and GFAP in the retinal tissue.Compared with the db/m group,the db/db,db/db+NC-SerpinA3N,and db/db+AAV-SerpinA3N groups showed decreased retinal thickness and RGC count,and increased number of TUNEL-positive cells,relative GFAP-positive expression intensity,levels of all inflammatory factors,and expression lev-els of NF-κB and Spi1 proteins,while SerpinA3N protein expression was decreased(all P＜0.05).Compared with the db/db group,the db/db+AAV-SerpinA3N group showed increased retinal thickness and RGC count,and decreased number of TUNEL-positive cells,relative GFAP-positive expression intensity,levels of all inflammatory factors,and expression levels of NF-κB and Spi1 proteins,while SerpinA3N protein expression was increased(all P＜0.05).Compared with the db/db+AAV-SerpinA3N group,the db/db+NC-SerpinA3N group showed decreased retinal thickness and RGC count,and increased number of TUNEL-positive cells,relative GFAP-positive expression intensity,levels of all inflammatory factors,and ex-pression levels of NF-κB and Spi1 proteins,while SerpinA3N protein expression was decreased(all P＜0.05).The PPI re-sults indicated an interaction between SerpinA3N and Spi1.Molecular docking results showed that Spi1 could form hydro-gen bonds with residues surrounding SerpinA3N.Conclusion Overexpression of SerpinA3N can inhibit Müller cell in-flammation and ameliorate retinal neural injury in diabetic mice,and the mechanism may be associated with the inhibition of the Spi1/NF-κB signaling pathway.
