Caffeine intake can have numerous effects on neurological and cognitive impairments. However, its influences on offspring still need to be evaluated. This study aimed to investigate the effects of caffeine exposure during gestation on behavioral development, metabolic function, and circadian rhythms in offspring. A total of 96 Sprague-Dawley rats were included, with an equal number of males and females. After confirming pregnancy, the female rats were randomly divided into four groups on average and administered caffeine at three distinct doses (5, 20, and 80 mg/kg/day) or equivalent volumes of normal saline throughout the gestational period. The health status and weight gain of pregnant rats, as well as the litter size and body weight of offspring, were continuously monitored. Open field test, Morris water maze, intraperitoneal glucose tolerance tests (IPGTT) and intraperitoneal insulin tolerance tests (IPITTs) were performed. The expression levels of 7 circadian clock genes (Bmal1, Clock, Cry1, Cry2, Per1, Per2, and Rev-erbα) were quantified. This revealed a significant reduction in maternal weight gain in the 80 mg/kg caffeine group compared to controls. Although the birth weight of offspring was not significantly affected, the postnatal weight gain of offspring was slower in caffeine groups. Behavioral assessments demonstrated elevated spontaneous activity and increased anxiety-like behaviors in caffeine-exposed offspring. Metabolic evaluations indicated impaired glucose tolerance specifically in female offspring, though no significant alterations were observed in insulin sensitivity, spatial learning and memory performance. Maternal caffeine exposure during gestation also disrupted the rhythmicity of hepatic circadian clock gene expression in offspring rats. These findings suggest that gestational caffeine exposure exerts selective adverse effects on offspring development, behavior, glucose metabolism and circadian rhythm. Caffeine consumption during pregnancy necessitates careful consideration.
