ORIGINAL ARTICLE The effects of caffeine on ovarian tissue in rats Maryam Yadegari Dehnavi 1,2 & Aghdas Mirjalili 1,2 & Hengameh Dortaj 1,3 & Abolghasem Abbasi Sarcheshmeh 1,2 & Fatemeh Zare Mehrjerdi 1,4 Received: 2 January 2020 /Accepted: 20 October 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020 Abstract Caffeine consumption increases during early adulthood, which has adverse effects on the reproductive system. This study aimed to assess the impact of embryonic caffeine exposure on rat ovary in adulthood. Female Wistar rats (240–270 g) were divided into 5 groups (n =7): experimental groups were exposed to 26, 45, 100, and 150 mg/kg of caffeine via drinking water during pregnancy and the control group only received drinking water. The ovaries of the offspring were taken out on days 7, 14, 28, 60, 90, and 120 of postnatal development, and then, they were fixed in 10% formaldehyde solution. Ovarian follicles were studied using stereological methods, and data were analyzed using one-way ANOVA followed by the Tukey test in SPSS software. A value of p < 0.05 was considered significant. The body weight, the weight of the ovaries, the ovarian volume, and the number of primordial follicles decreased significantly (p < 0.05) in 45 and 100 mg/kg, and (p < 0.001) in 150 mg/kg caffeine- treated groups at all stages of postnatal development. Significant decreases were observed in the number of primary and secondary follicles in 45 and 100 mg/kg (p < 0.05) and (p < 0.001) in 150 mg/kg caffeine-treated groups on days 7, 14, 28, and 60 compared to the control group. The number of Graafian follicles also decreased significantly (p < 0.001) in 45, 100, and 150 mg/kg caffeine-treated groups on days 14 and 28. Moreover, the mean volume of the oocyte in Graafian follicles reduced considerably in 45, 100, and 150 mg/kg caffeine-treated groups compared to other groups (p < 0.05). The thickness of the zona pellucida (ZP) in the secondary follicles (p < 0.02) and Graafian follicles (p < 0.05) showed a significant reduction in 100 and 150 mg/kg caffeine-treated groups on the 14th, 28th, and 60th days. In conclusion, high-dose caffeine consumption during gestation affects all stages of ovarian follicle development in rat offspring. Keywords Caffeine . Ovary . Follicle . Stereology . Rat offspring Abbreviations cAMP Cyclic adenosine monophosphate Cdc2 kinase Cyclin-dependent protein kinase MAP kinase Mitogen-activated protein kinase H&E Hematoxylin and eosin ZP Zona pellucida Introduction Some chemical and biological agents induced irreversible ef- fects on the reproductive system and fertility (Dorostghoal et al. 2011; Mehranjani and Mansoori 2016). Caffeine is one of the available nutrients used extensively by adoles- cents and exists in coffee and tea as well as chocolate, colas, energy drinks, and certain medications (Al- Shoshan 2007; Dorostghoal et al. 2011). The total daily intake of caffeine from any source is estimated to be approximately 3–7 mg/kg/day to 200 mg/day (Barone and Roberts. 1996; Dorostghoal et al. 2011). Peak plas- ma level of caffeine is reached between 15 and 120 min after oral ingestion in humans. Concerns have been raised about the decrease in reproductive health since some studies have pointed to the adverse effects of caf- feine on the female and male reproductive system (Pollard et al. 1990; Rhee et al. 2015; Yadegari et al. 2016; Belali Kharaji et al. 2019). * Maryam Yadegari Dehnavi yadegarimy@yahoo.com 1 Herbal Medicine Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran 2 Department of Anatomy and Cell Biology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran 3 Department of Tissue Engineering and Applied Cell Science, School of Advanced Medical Science and Technologies, Shiraz, Iran 4 Department of Physiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran Naunyn-Schmiedeberg's Archives of Pharmacology https://doi.org/10.1007/s00210-020-02008-8