To prevent lifestyle related diseases, it is important to modify lifestyle behavior. The control of mental stress level and prevention of mental stress-related diseases have become one of the most important problems in Japan. To check mental stress level objectively during the early stage of stress-related diseases and determine appropriate coping methods, it is necessary to design a useful index for mental stress. Cortisol is a steroid hormone secreted by the adrenal cortex. This is an essential hormone to human survival, and plays a key role in adaptation to stress. In another review, we concluded that cortisol appears to be an adequate index for mental stress.However, lifestyle factors such as alcohol drinking, smoking, lack of exercise etc., are strongly associated with mental stress. Thus, in this review, we focus on the relationship between cortisol and lifestyle.The present findings suggested that lifestyle factors; smoking, alcohol drinking, exercise, sleep and nutrition are strongly associated with cortisol levels, and it may be impossible to determine whether alterations in cortisol levels are due to mental stress.It was suggested that those lifestyle effects on not only mental stress itself but also cortisol levels should be considered, when assessing mental stress by cortisol levels.