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This study aimed to investigate the anti-aging effects and underlying molecular mechanisms of reserve mesenchyme cells （RMCs） derived from the velvet antler of the sika deer on hydrogen peroxide-induced senescent human dermal fibroblasts （HDFs）. An HDF senescence model was established by treatment with 200 µmol·L-1 H₂O₂ and confirmed via SA‑β‑galactosidase staining. Senescent HDFs were then co‑cultured with RMCs， and subsequent assays evaluated cell proliferation， migration， superoxide dismutase （SOD） activity， and malondialdehyde （MDA） content. The 4D-DIA proteomics technology was used to identify differential proteins between co-culture medium and senescent HDF medium， and bioinformatics enrichment analysis was performed. Functional analyses revealed that， compared with the model group， RMCs co‑culture significantly enhanced HDFs proliferation and migration （P<0.01）， increased SOD activity （P<0.05）， and decreased MDA levels （P<0.05）. Using 4D‑DIA proteomics， 289 differentially expressed proteins were identified in the conditioned media of the co‑culture versus senescent HDF alone， with key molecules such as ATP1A1， COL7A1， IGF2R， and MMP3 showing significant expression changes. KEGG enrichment analysis highlighted the proteasome， PPAR signaling pathway， extracellular matrix-receptor interaction， and focal adhesion pathways as principal regulatory axes. These findings suggest that velvet-antler-derived RMCs from the sika deer alleviate oxidative stress-induced dysfunction in senescent HDFs. It was speculated that RMCs may exert their effects by enhancing proteasome‑mediated clearance of damaged proteins， activating PPARγ to suppress inflammation and oxidative stress， promoting COL7A1‑dependent dermal-epidermal junction reconstruction， and inhibiting MMP3‑mediated extracellular matrix degradation， thus maintaining HDF homeostasis and function. This work provides experimental evidence and a theoretical foundation for developing deer antler-derived secretome-based strategies for skin anti‑aging.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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