Synchronize SIRT1-Mediated Oxidative Stress Resolution and HIF-1α Deacetylation to Combat Age-Related Macular Degeneration

Background: Age-related macular degeneration (AMD) therapy faces challenges in simultaneously targeting oxidative stress and hypoxia-driven angiogenesis. While Gynostemma pentaphyllum is traditionally used for "ocular stagnation", its active component gypenosides (Gps) and their dual-pathway mechanisms remain unexplored. This study investigates how Gps modulate SIRT1-mediated dual deacetylation to address AMD multifactorial pathogenesis. Methods: Network pharmacology identified 99 overlapping targets between Gps and AMD, with SIRT1 as the core node. Molecular docking (-9.3 kcal/mol binding energy) and 100-ns molecular dynamics simulations (RMSD <0.09 nm) confirmed stable Gps-SIRT1 binding at TYR-325/ARG-440 residues. Functional studies included: hypoxia/oxidative-stressed ARPE-19 cells and laser-induced choroidal neovascularization (CNV) mice (n=8/group), assessing SIRT1/SOD1/HIF-1α pathways via qPCR, Western blot, and fluorescein angiography. Results: Gps uniquely activated dual SIRT1 functions: 1) oxidative stress resolution via SIRT1 upregulation (2.1-fold, P<0.01) and SOD1 deacetylation (activity↑2.1-fold, P<0.01), reducing IL-1β by 67% (vs. Resveratrol P<0.05); 2) hypoxic signaling blockade through HIF-1α deacetylation (↓58%, P<0.001), suppressing VEGF-A (0.38-fold↓) and ANGPT2 (0.45-fold↓). In vivo, Gps achieved 42.3% CNV area reduction – outperforming anti-VEGF monotherapy (28.7%, P<0.05) with strong SIRT1-efficacy correlation (r=0.89). Conclusions: This work first demonstrates that Gps synchronize SIRT1-mediated mitochondrial redox regulation and hypoxia response inhibition through differential deacetylation, structurally anchored by TYR-325/ARG-440 interactions. The dual mechanism (-9.3 kcal/mol binding stability) validates G. pentaphyllum’s traditional use while providing a blueprint for developing multi-target botanical drugs against AMD.