NAD⁺ flux is maintained in aged mice despite lower tissue concentrations

NAD⁺ is an essential coenzyme for all living cells. NAD⁺ concentrations decline with age, but whether this reflects impaired production or accelerated consumption remains unclear. We employed isotope tracing and mass spectrometry to probe age-related changes in NAD⁺ metabolism across tissues. In aged mice, we observed modest tissue NAD⁺ depletion (median decrease ∼30%). Circulating NAD⁺ precursors were not significantly changed, and isotope tracing showed the unimpaired synthesis of nicotinamide from tryptophan. In most tissues of aged mice, turnover of the smaller tissue NAD⁺ pool was modestly faster such that absolute NAD⁺ biosynthetic flux was maintained, consistent with more active NAD⁺-consuming enzymes. Calorie restriction partially mitigated age-associated NAD⁺ decline by decreasing consumption. Acute inflammatory stress induced by LPS decreased NAD⁺ by impairing synthesis in both young and aged mice. Thus, the decline in NAD⁺ with normal aging is relatively subtle and occurs despite maintained NAD⁺ production, likely due to increased consumption.