染料木素对性成熟前小鼠卵巢卵泡发生动力学的影响

Effects of Genistein on Ovarian Folliculogenesis in Presexual Mice

【目的】旨在探究染料木素对性成熟前小鼠卵巢卵泡发生动力学的影响,为畜牧生产尤其是种猪饲养过程豆粕型日粮的合理饲喂以及黄酮类替抗产品的科学应用提供参考。【方法】将40只21日龄昆明小鼠随机分为4组,其中对照组处以0.1 mL的溶剂(含5%DMSO的玉米油),试验组分别处以等体积不同浓度的染料木素(25 mg/kg、50 mg/kg和100 mg/kg),连续腹腔注射7 d。处理结束后,对部分小鼠进行PMSG-HCG促排并记录输卵管壶腹部的卵子数量,评估卵巢排卵潜能;剩余小鼠采集卵巢制成组织切片进行HE染色,统计分析卵巢内各级卵泡生长和闭锁状态。【结果】输卵管壶腹部成熟卵子数统计显示,染料木素处理组的卵子数量较对照组均显著增加(P<0.05),其中50 mg/kg染料木素组卵子排出数量最多,且显著高于25 mg/kg和100 mg/kg剂量组(P<0.05),而25 mg/kg和100 mg/kg剂量组之间差异不显著(P>0.05)。卵泡计数结果显示,染料木素处理组生长卵泡总数较对照组均无显著差异,但健康生长卵泡显著高于对照组(P<0.05),闭锁生长卵泡则反之(P<0.05)。另外,3个剂量的染料木素处理组中正常原始卵泡的数量均高于对照组,且50 mg/kg和100 mg/kg剂量组较对照组增加显著(P<0.05);而异常原始卵泡的数量在染料木素处理组中均显著下降(P<0.05)。再者,50 mg/kg和100 mg/kg染料木素处理组的多卵母卵泡数量显著高于对照组和25 mg/kg剂量组(P<0.05)。【结论】染料木素以剂量依赖性方式抑制原始卵泡消亡和生长卵泡闭锁,促进生长卵泡发育成熟,从而提高卵巢排卵潜能,其中以50 mg/kg剂量的染料木素效果最佳。

[Objective]The purpose of this study was to explore the effects of genistein on ovarian folliculogenesis in presexual mice,which would provide a reference for the rational feeding of soybean meal diet and the scientific application of flavonoids substitute antibiotic products in livestock production,especially in pig production.[Method]Forty 21-day-old Kunming mice were randomly divided into four groups.One group was treated with 0.1 mL solvent(corn oil containing 5%DMSO)as the control,the other three groups were treated with 0.1 mL genistein at different concentrations(25 mg/kg,50 mg/kg and 100 mg/kg respectively).After 7 days of continuous intraperitoneal injection,a few mice were ovulated by PMSG-HCG and collected mature oocytes in the ampulla of fallopian tube to assess the ovulatory potential of their ovaries,whereas the other part of mice were used to harvest their ovaries for making tissue sections by HE staining.The follicular development and atresia status in the ovaries were then statistically analyzed.[Results]The numbers of mature oocytes in the oviductal jugular showed a significant increase in the genistein-treated groups compared with the control group(P<0.05),and the 50 mg/kg group had significantly higher number of oocytes than that of 25 mg/kg and 100 mg/kg groups(P<0.05).There was no significant difference between the 25 mg/kg and 100 mg/kg groups(P>0.05).The results of follicle counts showed that the total number of growing follicles in the genistein-treated groups were not significantly different from that of the control group,but the number of healthy growing follicles were all significantly higher than that of the control group(P<0.05),while the number of atretic growing follicles showed an opposite trend(P<0.05).In addition,the numbers of normal primordial follicles in 50 mg/kg and 100 mg/kg groups were significantly higher compared with the control group(P<0.05),while the number of abnormal primordial follicles decreased significantly in all genistein-treated groups(P<0.05).Furthermore,the number of multi-oocyte follicle in the 50 mg/kg and 100 mg/kg genistein-treated groups were significantly higher than that of the control group and the 25 mg/kg group.[Conclusion]The genistein inhibits primordial follicle extinction and growth follicle atresia in a dose-dependent manner and promotes the development and maturation of growth follicles,thereby potentially increasing ovarian ovulatory.Our findings also revealed that the dose of 50 mg/kg of genistein has the highest effective on mouse ovarian folliculogenesis.