Title : Effects of metformin, mitochondrial supplements and Hyperbaric Oxygen Therapy (HBOT) on biological aging biomarkers in aged BALB/c mice
Abstract:
Background: Aging is driven by interconnected hallmarks including telomere attrition, mitochondrial dysfunction, cellular senescence, and chronic low-grade inflammation that collectively reduce tissue resilience and regenerative capacity. Metformin, mitochondrial-support supplements (CoQ10, NMN, PQQ), and Hyperbaric Oxygen Therapy (HBOT) have been proposed as interventions targeting these pathways; however, the biological impact of short-duration exposure, and whether combining metabolic and oxygen-based strategies yields added benefit without compromising safety, remains uncertain. This study investigated whether these interventions, alone or in combination, modulate key biological aging biomarkers in aged BALB/c mice.
Methods: Aged female BALB/c mice (37–38 weeks) were allocated to five groups: control; metformin (100 mg/kg/day); supplement cocktail (CoQ10, NMN, PQQ); HBOT (100% O? at 1.53 ATA for 90 min/day); and a combined-treatment group. Interventions were administered for 30 days. Telomere length was quantified by qPCR. Senescence-associated gene expression (p16, p21, MMP12, MCP-1) was measured by qRT-PCR across tissues. Serum inflammatory cytokines (IL-1β, IL-6, IL-10, TNF-α, CRP) were assessed by ELISA. Histopathological evaluation of liver, kidney, lung, heart, and spleen was performed using H&E staining.
Results: No statistically significant changes in telomere length were observed in any intervention group compared with controls. Inflammatory cytokines demonstrated variable, non-significant fluctuations. Senescence gene expression showed organ-specific variability without consistent suppression of senescence pathways. Histological assessment did not demonstrate structural improvement or reversal of aging-associated tissue changes. Importantly, no safety concerns or adverse tissue responses were detected with single or combined interventions over the 30-day period.
Conclusion: In this aged mouse model, a 30-day course of metformin, mitochondrial supplements, HBOT or their combination did not significantly alter major biological aging markers. The heterogeneity across organs suggests differential tissue responsiveness and supports the need for longer treatment durations, dose/pressure optimization, and incorporation of more sensitive functional or stem cell–related assays to better capture regenerative and anti-aging effects.
