Human skin aging is a complex process driven by environmental stressors, most notably ultraviolet-B (UVB) radiation, which triggers the generation of reactive oxygen species (ROS). This oxidative stress leads to the upregulation of proteases like elastase and collagenase, which dismantle the extracellular matrix, resulting in wrinkles and loss of skin resilience. Vitex trifolia, an Indonesian medicinal plant traditionally used for skin disorders, was considered a potential solution due to its high content of polyphenols and flavonoids. These bioactives offer a multi-targeted approach, acting simultaneously as radical scavengers and enzyme inhibitors to provide a rational basis for natural cosmeceutical development.
Methods
Researchers utilized microwave-assisted extraction with 96% ethanol to obtain leaf extracts, which were characterized using LC–HRMS and phytochemical quantification. The study integrated in silico molecular docking to predict binding affinities with in vitro biochemical assays evaluating antioxidant (DPPH, ABTS, FRAP) and anti-protease activities. Finally, HaCaT keratinocytes were used to assess the extract’s cytotoxicity and cytoprotective effects against UVB damage, followed by qRT-PCR analysis of MMP-1 and MMP-9 gene expression.
Key Findings
• Phytochemical Profile: The extract is rich in phenolic compounds (78.52 mg GAE/g), with the methylated flavonoid casticin identified as the predominant bioactive metabolite.
• Antioxidant Potency: It demonstrated robust radical-scavenging activity across multiple mechanisms, achieving an IC50 of 63.47 µg/mL in DPPH assays.
• Enzyme Inhibition: The extract showed superior anti-collagenase activity (IC50 = 27.94 µg/mL) and moderate elastase inhibition, which was corroborated by favorable docking scores for compounds like quercetin and cyclohexane-derived phenolic esters.
• Cellular Safety and Protection: Unlike ascorbic acid, V. trifolia remained non-cytotoxic up to 100 µg/mL and provided dose-dependent cytoprotection against UVB-induced injury at concentrations of 12.5–50 µg/mL.
• MMP Regulation: Pre-treatment with the extract significantly downregulated UVB-induced MMP-1 and MMP-9 expression by 42% and 69%, respectively, outperforming the standard antioxidant ascorbic acid.
This research provides the first integrated evidence—bridging computational and cellular models—for the use of Vitex trifolia as a multifunctional anti-photoaging agent. Its novelty lies in its ability to target skin aging at multiple fronts, including oxidative stress, matrix degradation, and gene expression, while maintaining a broader safety profile than single-compound antioxidants. Future implications involve the translation of these findings into topical cosmeceutical formulations, with further research needed to explore in vivo efficacy and bioavailability.
Link to the study: https://www.mdpi.com/1422-0067/27/4/1913
Image of the below: Molecular docking interactions of selected compounds with MMP-9 and elastase. (a) Three-dimensional binding diagram (Red represents oxygen. Dark blue represents nitrogen. Light blue is carbon) and (b) three-dimensional conformation of compound 7 docked to MMP-9 (PDB ID: 6ESM). (c) Two-dimensional binding diagram and (d) three-dimensional conformation of quercetin docked to elastase (PDB ID: 2FOE). (a) Three-dimensional binding diagram and (b) three-dimensional conformation of compound 7 docked to the MMP-9 protein (PDB ID: 6ESM).
