BACKGROUND: While romantic jealousy may help to maintain relationships, following partner infidelity and an irretrievable loss of trust it can also promote break-ups. The neuropeptide oxytocin can enhance the maintenance of social bonds and reduce couple conflict, although its influence on jealousy evoked by imagined or real infidelity is unclear. AIMS: This study aimed to investigate the effects of intranasal oxytocin (24 IU) on romantic jealousy in both males and females in imagined and real contexts. METHODS: Seventy heterosexual couples participated in this double-blind, placebo-controlled, between-subject design study. Jealousy was firstly quantified in the context of subjects imagining partner infidelity and secondly in a Cyberball game where their partner interacted preferentially with an opposite-sexed rival stranger to simulate partner exclusion, or rejected a neutral stranger but not the partner. RESULTS: Oxytocin primarily decreased jealousy and arousal ratings towards imagined emotional and sexual infidelity by a partner in both sexes. During the Cyberball game, while male and female subjects in both groups subsequently threw the ball least often to the rival stranger, under oxytocin they showed reduced romantic jealousy and arousal ratings for stranger players, particularly the rival one, and reported reduced negative and increased positive feelings while playing the game. CONCLUSIONS: Together, our results suggest that oxytocin can reduce the negative emotional impact of jealousy in established romantic partners evoked by imagined or real infidelity or exclusive social interactions with others. This provides further support for oxytocin promoting maintenance of relationships.