Ageing is a complex and multifactorial process resulting in several functional and aesthetic changes to the skin. We found that α‐Naphthoflavone (α‐NF) concentration‐dependently induced pro‐collagen type I protein expression and inhibited MMP‐1 protein expression, in both normal and UVB‐irradiated cells. SB431542 and SIS3 – inhibitors of TGF‐β and Smad3, respectively – significantly alleviate α‐NF‐caused response of MMP‐1 and pro‐collagen. LY294002 (PI3K inhibitor) can reverse α‐NF‐induced ERK, Akt, Smad‐3 activation, pro‐collagen synthesis and α‐NF‐suppressed AP‐1 activation. PD (ERK inhibitor) was not involved in pro‐collagen generation and MMP‐1 inhibition. We concluded that α‐NF promotes pro‐collagen production and inhibits MMP‐1 expression via the activation of a PI3K/Akt/Smad‐3 pathway in normal and UVB‐irradiated human skin fibroblasts, while TGF‐β may play an important role in transducing this pathway. These results suggest that α‐NF, a natural plant product, has the potential to become a novel anti‐ageing skin application.