Couple conflict in unhappy marriages is suggested to impair individual health via chronic psychophysiological stress reactions in couples' everyday lives. As a consequence, we hypothesized that standard couples relationship education (CRE) would decrease psychophysiological stress, namely salivary cortisol levels, during couple conflict in the laboratory as compared to a standard psychological stress paradigm. We considered cortisol to be of particular interest in this context, as it mediates endocrine and immune responses to stress, and thus might influence couples' health. Salivary cortisol was repeatedly investigated in 61 couples during (a) a standard psychological stress test with no relevance for the couples, and (b) a standard couple conflict discussion in the laboratory before and after CRE. In addition, increases in self-evaluated relationship quality were analyzed with regard to their influence on salivary cortisol. Data were analyzed using multilevel modeling. Cortisol responses to the couple-external psychological stress test were unaffected by CRE, but specifically cortisol responses during the couple conflict discussion were significantly reduced following CRE compared to pre-intervention levels. Moreover, cortisol decreases during conflict were partially mediated by increases in self-reported relationship quality following CRE. These data suggest that CRE might buffer the harmful effects of repeated conflict in close relationships. Rather than ameliorating overall stress resilience, CRE might thus specifically improve individual health through increased relationship quality and reduced HPA axis activity during couple conflict.