There is evidence to suggest that microgravity/weightlessness can induce changes in lung physiology/function. We hypothesized that microgravity, induced by head-down bed rest (HDBR), would induce changes in lung function and that exercise training with artificial gravity (AG) would prevent these changes from occurring. Twelve participants were randomly assigned to a control or AG exercise countermeasure (CM) group (n = 6 per group) and 96 h of 6° HDBR. Participants in the CM group were exposed to AG (alternating 2 min intervals of +1·0 and +2·0 G) for 30 min, twice daily, during which time ergometric exercise (40 W intensity) was performed. Pulse rate, oxygen saturation (SO2) and lung function were measured and compared between groups. The CM and control groups were similar in mean age, height and weight. There were no significant within or between group differences over time in pulse rate, SO2, vital capacity, inspiratory capacity, tidal volume, expiratory reserve volume, inspiratory reserve volume, minute ventilation, forced vital capacity, forced expiratory volume in 1 s, peak expiratory flow, maximal expiratory flow in 25%, 50% and 75% vital capacity, forced inspiratory vital capacity, forced inspiratory volume in 1 s and maximal voluntary ventilation. Microgravity induced by 96 h of HDBR does not appear to affect lung function in humans. Further, AG with exercise training does not change lung function during 96 h of HDBR in humans.