Blueberry Anthocyanins Alleviate Testicular Aging in Middle‐Aged Mice by Targeting Inflammaging and Reprogramming the cAMP–Wnt and Methionine–NAD + Metabolic Axes

ABSTRACT Emerging evidence suggests that inflammaging contributes significantly to male reproductive senescence and oxidative stress, yet safe, non‐invasive interventions that act on the upstream regulatory circuitry of the spermatogonial niche remain elusive. Here we investigated the restorative potential of a high‐anthocyanin Blueberry Anthocyanin Extract (BAE) in a middle‐aged murine model, integrating physiological monitoring, histopathology, and paired testicular transcriptomics and metabolomics to resolve the molecular logic of BAE‐mediated rejuvenation. BAE prevented age‐associated physiological deterioration without detectable toxicity, reversed seminiferous tubule degeneration, and restored sperm kinematics. Mechanistically, multi‐omics integration converged on three coordinated regulatory modules rather than a single pathway. First, BAE engaged adenylyl cyclase–cAMP–PKA signaling to license canonical Wnt /β‐catenin activity, driving spermatogonial proliferation (PLZF, PCNA, c‐Myc , Ccnd1 ). Second, BAE suppressed ROS‐dependent IKK/NF‐κBNF (p65) activation, lowering testicular and systemic TNF‐α and IL‐6 and resolving the pro‐inflammatory niche. Third, BAE reprogrammed the methionine cycle and elevated NAD + , activating the AMPK–SIRT1–PGC1α axis while restraining leucine‐ and methionine‐driven mTORC1 signaling, thereby coupling redox balance to cell‐cycle re‐entry. Collectively, these data identify BAE as a dual‐action, immunometabolic modulator that simultaneously reactivates the spermatogonial proliferative program and dismantles the inflammaging circuit. By concurrently targeting the cAMP–Wnt, ROS–NF‐κB, and methionine–NAD + axes, high‐anthocyanin BAE offers a mechanism‐based functional‐food strategy to delay male reproductive aging and preserve fertility.