Influence And Mechanism Discussion Of Isoflurane Exposure On Female Mice Ovarian Function

PART I Influence of 15-Day-Consecutive Isoflurane Exposure on Female Ovarian FunctionObjective To investigate whether isoflurane exposure may negatively influence adult female ovarian functionMethods A 15-day-consecutive exposure model is introduced to explore the impact of isoflurane on adult female C57 mice ovarian function.One hundred adult female C57 mice were randomly allocated into 5 groups with atmosphere containing 0,2500,5000,10000 and 20000 ppm isoflurane inhaled.Estrous cycle,follicle counting and serum hormones were performed to assess potential ovarian-toxicity of isofluraneResults Prolonged adult female mice estrous cycle were observed in 10000,20000ppm groups,but only significant extension of estrous phase in 20000ppm group.Serum hormone fluctuations(AMH,FSH and LH level)were remarkable in 5000,10000 and 20000ppm groups.Additionally,reduced developing follicles as well as increased atresia follicles recognized after isoflurane exposure but with no statistical significanceConclusion Thus we concluded higher concentration isoflurane exposure interfered physiological ovulation and sexual hormone endocrines,which reminded us of adverse effect of the drug on ovarian reserve in sexual-mature female micePART Ⅱ Mechanism Discussion of Isoflurane-Induced Ovarian Function DeclineObjective To investigate isoflurane-induced oxidative stress fluctuations,apoptosis-associated molecules expressions and related mitochondrial function alternations in granulosa cells.Methods The ROS measurement was introduced to evaluate oxidative stress in C57 mice primary granulosa cells after isofurane exposure(2%isoflurane plus 21%O2 and 5%CO2.)or controlled environment exposure(21%O2 and 5%CO2.)for 6 hours.In avoid of cell death,granulosa cells were exposed in 3-hour isoflurane environment(2%isoflurane plus 21%O2 and 5%CO2.)or 3-hour controlled environment exposure(21%02 and 5%C02)for cellular ATP level assessment.Moreover,ratio of bax,bcl2 were analyzed in transcriptional and translational levels in both-group granulocytes after 6-hour exposure in respected environmentResults Compared with control,abnormally increased ROS level was observed in isoflurane-exposed granulocytes,significant decline of ATP production was also noticed in granulocytes after 6-hour-isoflurane exposure.Moreover,up-regulated bax/bcl2 in transcriptional and translational levels were identified in isoflurane-exposed granulocytesConclusion Isoflurane possibly promoted granulocytes apoptosis through abnormally overloading ovarian granulosa cells with excess oxidative stress,activation of bax,bcl2 mitochondrial-apoptosis pathway and ultimately deprivation of cellular energy.