Tu-Si-Zi-Wan reduces D-galactose-induced hepatic and cerebral oxidative damage in aging mice via the Nrf2/ARE pathway.

The global aging population necessitates safe, multi-target strategies to combat age-related functional decline. Tu-Si-Zi-Wan (TSZW), a classical medicinal-dietary formula documented in ancient Chinese medical texts and composed of Cuscuta chinensis and Dioscorea opposita, demonstrates multitargeted potential, yet its systemic antiaging mechanisms remain unclear. This study investigated the protective effects of TSZW on hepatic and neurological aging through the Nrf2/ARE pathway while exploring its systemic impact on metabolic and immune homeostasis. Firstly, network pharmacology predicted the targets and pathways of TSZW, identifying 113 shared TSZW-aging targets, with Nrf2, IL-6, and HIF-1α as core nodes enriched in oxidative stress-related pathways. Secondly, a D-galactose-induced aging mouse model was established, followed by 4 months of TSZW intervention (low, medium, and high doses). Behavioral tests (open field, novel object recognition, and grip strength), metabolic and inflammatory marker detection, histopathology, and molecular analyses (western blot, quantitative PCR, and immunofluorescence) were used to assess systemic aging phenotypes, oxidative damage, and Nrf2/ARE pathway activity. TSZW dose-dependently improved motor activity, cognition, and muscle strength; restored liver/kidney function (ALT, AST, Cr, and urea); reduced the serum levels of tumor necrosis factor-α, interleukin 6, and interleukin-1β; and attenuated hepatic/neuronal damage. It suppressed the expression of senescence markers (p16, p21, and p53) while increasing the expression of antioxidant enzymes (SOD and GSH-Px) and reducing oxidative damage (MDA and ROS). Mechanistically, TSZW promoted Nrf2 nuclear translocation; upregulated Hmox1, Nqo1, and Gclm expression; and inhibited Keap1. Notably, high-dose TSZW outperformed vitamin E in reversing immune organ atrophy (thymus and spleen indices) and preserving hippocampal neurons. Collectively, TSZW alleviates hepatic and neurological aging via Nrf2/ARE-mediated antioxidant signaling, with concurrent mitigation of systemic aging phenotypes, including immune organ atrophy and chronic inflammation. Its multicomponent synergy highlights Traditional Chinese medicinal cuisine's potential for systemic anti-aging intervention by targeting oxidative stress-inflammation-metabolic signaling crosstalk.