...Here, we showed that blood donation increased dermal thickness and collagen content and decreased the number of senescent cells in old mice. Transcriptomic and metabolomic studies revealed blood donation significantly altered aging-related pathways in the skin of old mice. Molecular genes analysis indicated blood donation decreased the expression of genes associated with inflammation such as Fols1, Cox-2, and IL-1β, and increased the expression of collagen-associated genes including TGF-β1, TGF-β2, and Col3a1. The improvement of skin aging by blood donation was associated with the reduction of iron deposits and the increase of TGF-β1 in elderly skin. Our results suggested that appropriate blood donation could promote collagen re-synthesis and improve skin aging.