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Danggui Buxue Decoction Attenuates H2O2-Induced HaCaT Cells by Modulating SOD2 Antioxidant and P21 Senescence
Xingyu Chen1, Huilan Zheng2, Hongbin Cheng1
1Department of Dermatology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan, 610075, People's Republic of China.
Background:
Skin aging is linked to oxidative stress, and functional decline of keratinocytes is a core feature. Common chemical anti-aging agents, such as retinoic acid, often cause skin irritation, whereas traditional Chinese medicine formulas, with multiple components and relatively low toxicity, may offer alternative strategies.
Methods:
To investigate its antiaging effects and mechanisms, an in vitro oxidative stress model was established using H2O2-treated human HaCaT cells. The cellular safety of Danggui Buxue Decoction was assessed by CCK-8 assay and cell morphology. The effects of pretreatment on cell viability, intracellular reactive oxygen species, superoxide dismutase and glutathione peroxidase activities, and malondialdehyde content were evaluated. Real-time quantitative PCR was used to determine the mRNA expression of senescence- and oxidative stress-related genes.
Results:
Exposure to 600 μmol/L H2O2 for 1 h established an oxidative stress model, reducing cell viability to 66.3%, increasing ROS and MDA levels, and decreasing SOD and GSH-Px activities. Danggui Buxue Decoction showed no cytotoxicity at 6.25-1600 μg/mL. Pretreatment with 400 μg/mL Danggui Buxue Decoction provided protection against oxidative damage, improving cell viability, lowering ROS and MDA levels, and restoring SOD and GSH-Px activities. Specifically, 400 μg/mL Danggui Buxue Decoction increased cell viability by 31.7%, enhanced SOD and GSH-Px activities by 91.3% and 32.2%, respectively, and reduced MDA content by 27.9%. Gene expression analysis indicated that pretreatment prevented the H2O2-induced decrease in SOD2 and MMP1 expression and attenuated the H2O2-induced overexpression of PPARα and P21.
Conclusion:
Danggui Buxue Decoction exerts preventive antiaging protective effects on HaCaT cells by scavenging excess reactive oxygen species, preventing impairment of the endogenous antioxidant enzyme system, inhibiting lipid peroxidation, and regulating key genes such as SOD2, MMP1, PPARα, and P21, thereby interrupting the vicious cycle of oxidative stress-induced cellular senescence and supporting its potential as a natural anti-aging agent. In vivo and clinical studies are needed to validate the translational applicability of these findings in the future.
Insights
Danggui Buxue Decoction protects skin cells from oxidative stress and aging. This traditional Chinese medicine enhances antioxidant defenses and regulates key aging genes, showing potential as a natural anti-aging agent.
Area of Science:
- Dermatology
- Cell Biology
- Traditional Chinese Medicine
Background:
- Skin aging is associated with oxidative stress and keratinocyte dysfunction.
- Conventional anti-aging chemicals can cause irritation.
- Traditional Chinese Medicine offers potential alternatives with lower toxicity.
Purpose of the Study:
- To investigate the anti-aging effects and mechanisms of Danggui Buxue Decoction (DBD) on human keratinocytes under oxidative stress.
- To establish an in vitro model of oxidative stress using hydrogen peroxide (H2O2)-treated HaCaT cells.
Main Methods:
- An in vitro oxidative stress model was created using H2O2-treated HaCaT cells.
- Cellular safety of DBD was assessed using CCK-8 assay and morphology.
- Effects on cell viability, reactive oxygen species (ROS), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) were evaluated.
- mRNA expression of senescence and oxidative stress-related genes was analyzed via real-time quantitative PCR.
Main Results:
- H2O2-induced oxidative stress reduced cell viability and antioxidant enzyme activity while increasing ROS and MDA levels.
- DBD showed no cytotoxicity at concentrations up to 1600 μg/mL.
- DBD pretreatment (400 μg/mL) protected cells by improving viability, reducing ROS and MDA, and restoring SOD and GSH-Px activities.
- DBD regulated gene expression, preventing decreases in SOD2 and MMP1 and attenuating increases in PPARα and P21.
Conclusions:
- Danggui Buxue Decoction demonstrates preventive anti-aging effects on HaCaT cells.
- It scavenges ROS, preserves antioxidant enzyme systems, inhibits lipid peroxidation, and modulates key aging-related genes.
- DBD shows potential as a natural anti-aging agent, warranting further in vivo and clinical investigation.
