关键词: 红石榴提取物, 牡丹花提取物, 网络药理学, 晚期糖基化终末产物, 皮肤衰老, AGE-RAGE/TGF-β1信号通路

Abstract:

The research aims to explore the anti-skin aging effect and mechanism of action of the red pomegranate-peony flower extract (RPP) through network pharmacology, molecular docking and experimental verification. Ultra-high performance liquid chromatography-tandem mass spectrometry was employed to identify the main bioactive components of RPP. Potential gene targets associated with skin glycation were screened based on the TTD and GeneCards database, and core targets were filtered via protein-protein interaction network analysis. Further,GO and KEGG pathway enrichment analysis were performed on the core targets to construct a multi-level "bioactive component-target-pathway" network, and molecular docking technology was used to verify the binding ability between bioactive components and key targets. Zebrafish oxidative stress models were induced by tert-butyl hydroperoxide and menadione K₃ respectively, and reactive oxygen species levels and relative opaqueness levels were detected; an in vitro glycation model was established, and anti-glycation activity was evaluated by biochemical index detection; a Caenorhabditis elegans glycation model was induced with glucose-fructose mixed solution to assess its effect on glycation-related phenotypes; a human dermal fibroblast glycation model was induced by methylglyoxal to detect glycation damage and intervention effects at the cellular level.Network pharmacology and molecular docking predict that RPP may slow down the advanced glycation end products – receptor for advanced glycation end products (AGE-RAGE) signaling pathway reduce the fibrosis of the extracellular matrix, regulate the expression of matrix metalloproteinase (MMP), and thereby prevent the degradation of collagen and elastin in the extracellular matrix, thereby slowing down skin aging. The experimental results show that RPP can significantly reduce the accumulation of reactive oxygen species in zebrafish (P < 0.001), and can significantly inhibit the production of pigment in the yolk sac of zebrafish induced by oxidative stress (P < 0.05), demonstrating an antioxidant effect. RPP can also inhibit the generation of advanced glycation end products (inhibition rate is 62.25%±0.17%), significantly reduce the content of pentose in Caenorhabditis elegans (P < 0.001) and increase the expression level of collagen XII(col-12) mRNA (P < 0.05); It significantly reduced the content of pentose glycoside and carboxymethyl lysinase in human skin fibroblasts (P < 0.001) and had an anti-skin glycation effect. RPP exerts anti-skin aging effects by regulating the AGE-RAGE/ transforming growth factor-beta 1(TGF-β1) signaling pathway, synchronously inhibiting the expressions of MMP1 and MMP3, maintaining extracellular matrix homeostasis, and reducing oxidative stress and glycation damage.

Key words: red pomegranate extract; peony flower extract, network pharmacology, advanced glycation end products; skin aging, AGE-RAGE/TGF-β1 signaling pathway