DHEA诱导小鼠PCOS模型过程中性激素的变化规律

Changes of Sex Hormone Levels in the Process of PCOS Induced by DHEA in Mice

【目的】鉴于在多囊卵巢综合征(polycystic ovary syndrome,PCOS)发病机理和治疗方式上的诸多争议,以及使用人类PCOS材料进行研究的局限性,本研究通过构建PCOS小鼠模型以探究在模型构建过程中性激素水平的变化规律。【方法】皮下注射脱氢表雄酮(dehydroepiandrosterone,DHEA)诱导昆明白雌鼠产生PCOS样临床症状,通过结晶紫染色查看小鼠性周期是否发生停滞;利用HE染色的方式确定卵巢发育状况;利用ELISA技术调查建模过程中血清睾酮和雌二醇浓度的变化规律。【结果】使用结晶紫对小鼠阴道上皮细胞染色可准确区分小鼠各个性周期阶段;使用6 mg/100 g DHEA持续诱导20 d后,昆明白小鼠性周期循环发生了一定的停滞,且在建模过程中体重变化与芝麻油溶剂的加入呈显著相关(P<0.05),与DHEA处理无关;DHEA连续处理后可见卵巢中巨大囊状卵泡出现,血清睾酮水平出现显著升高(P<0.05);自PCOS模型构建的第5天起,血清睾酮水平显著上升(P<0.05),且维持到建模结束;而血清雌二醇水平出现了阶段性变化,在第10、15天均显著高于对照组(P<0.05),第20天时低于对照组。【结论】改进DHEA溶解方式可成功构建昆明白小鼠PCOS模型,在建模过程中血清睾酮持续处于较高水平,而雌二醇水平呈现先增高后降低的趋势,后者在建模中期瞬时升高的病理意义需进一步研究。

【Objective】In view of the controversy over the pathogenesis and treatment of polycystic ovary syndrome(PCOS),and the limited use of human PCOS materials for research,in the present study,PCOS mice model was constructed for further exploring the changes in sex hormone levels in this process.【Method】Subcutaneous injection of dehydroepiandrosterone(DHEA)was chosen to produce KM female mice with PCOS-like clinical symptoms,mouse estrous cycle was identified by crystal violet staining.The ovarian development status was determined with HE staining.The changes of serum testosterone and estradiol concentrations during the modeling process were evaluated using ELISA method.【Result】The vaginal epithelial cells of mice were stained with crystal violet,which could accurately distinguish the different stages of estrous cycle in mice.After 20 days of continuous induction with 6 mg/100 g DHEA,the estrous cycle of KM mice was arrested,and the change of body weight was significantly related to the addition of sesame oil solvent(P<0.05),but there was no relation to DHEA treatment.After continuous treatment with DHEA,the serum testosterone level increased significantly(P<0.05),and the serum testosterone level increased significantly from the 5th day of PCOS model establishment and maintained until the end(P<0.05).The serum estradiol level was significantly higher than that of control group on the 10th and 15th day(P<0.05),and lower than that of control group on the 20th day.【Conclusion】The PCOS KM mouse model was successfully established by improving the DHEA dissolution method.During the modeling process,the concentration of androgen continued to be at a high level,while the concentration of estradiol increased at the early stage and decreased at the end of modeling,however,the possible pathological role on estradiol elevated instantly during the modeling process needed to be further verified.