滋阴明目方对干性年龄相关性黄斑变性模型小鼠视网膜的保护作用及机制OA

Objective To explore the protective effect of Ziyin Mingmu Formula(ZY)on mice with dry age-related macular degeneration(dAMD)and its underlying mechanism.Methods A mouse dAMD model was established using a high-fat diet combined with sodium iodate injection.Sixty mice were randomly divided into six groups:blank group,model group,ZY-low-dose group(ZY-L group),ZY-medium dose group(ZY-M group),ZY-high-dose group(ZY-H group),and Lu-tein group,with 10 mice in each group.Mice in the blank group were fed with regular feed for 3 months and then intraperito-neally injected with physiological saline,while mice in the other groups were fed with high-fat feed for 3 months and then intraperitoneally injected with sodium iodate(20 mg·kg-1).Mice in the ZY-L,ZY-M,and ZY-H groups were given ZY tradi-tional Chinese medicine solution at doses of 11,22,and 44 g·kg-1,respectively,by gavage once a day for 2 consecutive weeks,while mice in the blank group and the model group were treated with physiological saline by gavage.The retinal inju-ry of mice was evaluated using optical coherence tomography,fundus autofluorescence,and hematoxylin-eosin staining;dif-ferentially expressed genes in mouse eyeball tissue were analyzed using transcriptomics;the expression level of phosphoryl-ated ribosomal protein S6 kinase 1 in mouse eyeball tissue was detected using immunofluorescence staining;the expression levels of key inflammatory factor related proteins[interleukin-6(IL-6),monocyte chemoattractant protein-1(MCP-1),and matrix metalloproteinase-9(MMP-9)]in the serum of mice with age-related secretory phenotypes were measured using en-zyme-linked immunosorbent assay(ELISA);the expression levels of mTOR pathway related proteins in mouse retina,inclu-ding phosphorylated mammalian target of rapamycin(p-mTOR),p-p70S6K,tumor protein 53(P53),and cyclin dependent kinase inhibitor 1A(P21),were determined using Western blot.Results The imaging results showed that ZY intervention significantly alleviated retinal damage in the dAMD mouse model,reduced atrophy area,decreased verrucous deposition on the vitreous membrane,and improved retinal thinning.The transcriptomic analysis results showed that compared with the model group,there were 1 592 differentially expressed genes in the ZY-H group,and according to the KEGG enrichment analysis,they were mainly enriched in glycerophospholipid metabolism pathway,mTOR signaling pathway,glutathione metabolism pathway,adipocytokine signaling pathway,and vascular endothelial growth factor signaling pathway.The immu-nofluorescence staining results showed that compared with the model group,the immunofluorescence staining intensity of p-p70S6K decreased in both the ZY-H group and the Lutein group,and the differences were statistically significant(both P＜0.01).The ELISA results showed that compared with the model group,the expression levels of IL-6,MMP-9,and MCP-1 in the serum of mice in the ZY-H and Lutein groups decreased,and the differences were statistically significant(all P＜0.05).The Western blot results showed that compared with the model group,the expression levels of p-mTOR and p-p70S6K in the retinal tissue of mice in the ZY-H and Lutein groups decreased;the expression levels of P53 and P21 in the retinal tissue of mice in the ZY-M,ZY-H,and Lutein groups decreased,and the differences were statistically significant(all P＜0.05).Con-clusion ZY exerts a protective effect on the retina of the dAMD mouse model by inhibiting the mTORC1/p-70S6K path-way and reducing cell aging-related key factors such as P53 and P21,as well as aging-related secretory phenotypes.