Dendrobium nobile is a well-known traditional Chinese herbal medicine, and DNLA (Dendrobium nobile Lindl. alkaloids) is the active ingredient to have anti-brain aging effects in laboratory animals. The aim of the present study is to examine the anti-aging effects of DNLA in D-galactose induced primary hippocampal neuron senescence. Primary hippocampal neuron cultures were pretreated with DNLA (35 and 350 ng/mL) for 24 h, then exposed to D-galactose (50 mM) for 48 h. Cell viability was evaluated by the CCK-8 assay. Flow cytometry analysis was performed to evaluate the apoptosis rate of hippocampal neurons. β-Galactosidase staining was used to detect the senescence of neurons. Western blot analysis was used to detect the protein expression. The results show that DNLA markedly recovered the reduction in cell viability, inhibited the D-gal-induced hippocampal neuron apoptosis, decreased the senescence-associated β-galactosidase activity, and attenuated the protein expression of senescence marker p53 and p16 in D-galactose induced senescent hippocampal neurons. Furthermore, DNLA enhanced autophagy activity by increasing LC3-II/LC3-I ratio, Beclin1, and by decreasing p62 in D-galactose induced senescent hippocampal neurons. Moreover, DNLA upregulated the expression of autophagy-regulator SIRT1 and FoxO1. These novel findings demonstrate that DNLA is effective in protecting against D-galactose induced senescent hippocampal neurons in vitro. The anti-aging mechanisms of DNLA appear to be mediated, at least in part, through activated Sirt1-FoxO1-autophagy axis. © 2023 The Author(s)