SummaryXuanwei ham is a kind of traditional meat product with unique flavour along with abundant of bioactive peptides. This study was to investigate the in vitro stability of bioactive peptide extract from dry‐cured Xuanwei hams and its anti‐inflammatory effect in RAW264.7 cells. Firstly, the Xuanwei ham peptides (XHPs) were extracted and then tested in the lipopolysaccharide (LPS)‐induced macrophage cell model. Furthermore, the XHPs were exposed to different pretreatments including heating, acid‐base environment and simulated gastrointestinal digestion to evaluate their inflammatory regulation capacity. The heating temperature and pH did not show significant effects on the anti‐inflammatory capacity of XHPs, whereas the secretion of NO and TNF‐α was not suppressed after heating at 100 °C for 60 min. After digestion, the numbers of free amino acids and short peptides were enhanced significantly and the XHPs showed better inhibitory effect on LPS‐induced NO and TNF‐α secretion. Overall, the heating and pH treatment had no significant influence on the stability of XHPs but the anti‐inflammatory effect was enhanced after the simulated gastrointestinal digestion along with the releasing of novel bioactive peptides.