柠檬酸对小鼠卵泡组织形态的影响

摘要

通过观察小鼠卵泡形态结构的变化,研究柠檬酸对雌性小鼠卵泡发育的影响.将60只20～22 g昆明小鼠随机分为生理盐水对照组和柠檬酸低、中、高剂量处理组,各组小鼠每天每只依次分别腹腔注射0.2mL的0.9％生理盐水、7.5 mg/mL、15 mg/mL及30 mg/mL的柠檬酸.第21天在乙醚麻醉条件下,每组随机取10只小鼠断颈处死,摘取卵巢浸泡于4％的甲醛溶液中固定24 h,制作常规石蜡切片,HE染色,光学显微镜进行组织切片观察.结果表明:对照组小鼠各级卵泡的形态正常,7.5 mg/mL、15 mg/mL柠檬酸处理组可观察到正常形态的原始卵泡、初级卵泡,30 mg/mL柠檬酸处理组因原始卵泡、初级卵泡退化,使原始卵泡、初级卵泡的数量显著减少;15 mg/mL柠檬酸处理组主要使次级卵泡发育受阻,某些小的有腔卵泡部分颗粒细胞凋亡,卵泡膜细胞分化障碍,大的有腔卵泡的卵母细胞和卵泡内膜细胞退化,以及卵泡膜层血管数量减少或消失;30 mg/mL柠檬酸处理组卵母细胞发育周期阻滞或退化、消失,导致原始卵泡和初级卵泡数量显著减少,以及次级卵泡、有腔卵泡的闭锁.结果提示:低、中剂量的柠檬酸对原始卵泡、初级卵泡的发育无影响;中剂量柠檬酸使次级卵泡发育受阻,有腔卵泡退化;高剂量柠檬酸对各级卵泡的发育均有显著影响,造成卵泡的闭锁.%This research was conducted to investigate the effects of citric acid on various stages of development and morphology for follicles on female mice.60 Kunming female mice weighing 20-22 g were randomly divided into four groups including control group and low-dose,middle-dose,high-dose Citric acid (CA)-treated groups.Experiment group mice were respectively injected intraperitoneally with 0.2 ml CA ranging from 7.5,15 to 30 mg/ml once daily,and the same amount of physiological saline to control group.10 mice were randomly chosen from each group and were sacrificed at 21 d after injected decapitation under light ether anesthesia.The ovaries immersed in 4％ formaldehyde solution for 24 h,then was made into paraffin tissue section with HE stained and optical microscope observation.The results show that the morphology and quantity of follicles at different levels were normal in control group,the normal form of primitive follicles and primary follicles were observed for 7.5 mg/ml or 15 mg/ml citric acid group.Because of primordial follicle and primary follicles degraded,the number of primordial follicle or primary follicle significantly reduced in 30 mg/ml citric acid group,there were atresia on secondary follicle and antral follicle.It is concluded that low-dose and middle-dose citric acid don't affect development of original follicle and primary follicle,secondary follicle development were blocked by middle-dose citric acid and antral follicle cellular structure degradation.High-dose citric acid can significantly impact development of various follicle and cause follicular atresia.