双氢睾酮联合高脂饮食诱导多囊卵巢综合征样伴糖脂代谢异常的小鼠模型

A mouse model of polycystic ovary syndrome with abnormal glucose and lipid metabolism induced by dihydrotestosterone combined with high-fat diet

目的:联合双氢睾酮(dihydrotestosterone,DHT)和高脂饮食建立一种多囊卵巢综合征(polycystic ovary syndrome,PCOS)样伴糖脂代谢异常的小鼠模型。方法:将16只3周龄清洁级雌性C57BL/6小鼠随机分为2组干预6周:实验组(n=8)予皮下注射DHT+高脂饮食干预,对照组(n=8)予皮下注射有机溶剂+普通饮食干预。干预结束后比较2组小鼠动情周期、性激素水平、卵巢病理、体质量、血糖、血脂、皮下及内脏脂肪库等,判定模型成功与否。结果:干预完毕后,实验组血清DHT水平约为对照组的3倍(P=0.000)。连续观察14 d,实验组均无动情周期,而对照组均有规律动情周期。实验组动情间期明显长于对照组(P=0.001),而动情前期(P=0.006)和动情期(P=0.000)的持续时间均短于对照组。光镜下,实验组小鼠卵巢呈多囊性改变,颗粒细胞层数减少,膜细胞增生、厚度增加,未见黄体,而小窦卵泡(P=0.043)和大窦卵泡(P=0.000)数量明显增多,小窦卵泡中不健康卵泡比例(P=0.011)和大窦卵泡中不健康卵泡比例(P=0.001)也明显升高。实验组空腹血糖水平(P=0.001)、腹腔葡萄糖耐量实验(P=0.012)和腹腔胰岛素耐量实验(P=0.007)曲线下面积均明显大于对照组。实验组小鼠增加的体质量明显大于对照组(P=0.005),其腹膜后(P=0.007)、子宫旁(P=0.006)和腹股沟(P=0.018)脂肪库的质量,以及血清甘油三酯(P=0.009)和总胆固醇(P=0.000)均大于对照组。结论:联合DHT和高脂饮食可诱导小鼠卵巢多囊样改变及动情周期改变,伴发糖脂代谢异常,为PCOS发病机制的实验研究提供了一种较好的小鼠模型。

Objective:To establish a mouse model of polycystic ovary syndrome(PCOS)with abnormal glucose and lipid metabolism by combining dihydrotestosterone(DHT)and high-fat diet.Methods:Sixteen 3-week-old clean grade female C57BL/6 mice were randomly divided into two groups for intervention for 6 weeks:the experimental group(8 mice)was treated with subcutaneous injection of DHT + high-fat diet,and the control group(8 mice)was treated with subcutaneous injection of organic solvent + ordinary diet. After the intervention,the estrous cycle,sex hormone level,ovarian pathology,body mass,blood glucose,blood lipid,subcutaneous and visceral fat depot of the two groups were compared to determine the success of the model.Results:After the intervention,the level of serum DHT in the experimental group was about 3 times higher than that in the control group(P=0.000). After continuous observation for 14 days,the experimental group had no estrous cycle,while the control group had regular estrous cycle. The time of diestrus in the experimental group was significantly longer than that in the control group(P=0.001),while the duration of proestrous(P=0.006)and estrous(P=0.000)was shorter than that in the control group. Under light microscope,the ovaries in the experimental group showed polycystic changes,the layers of granulosa cells decreased,the proliferation and thickness of theca cells increased,no corpus luteum was found,the number of small antral follicles(P=0.043)and large antral follicles(P=0.000)increased significantly,and the proportion of unhealthy follicles in small antral follicles(P=0.011)and large antral follicles(P=0.001)also increased significantly. The fasting blood glucose level(P=0.001),area under the curve of IPGTT(P=0.012)and IPITT(P=0.007)in the experimental group were significantly larger than those in the control group. The weight increase of mice in the experimental group was significantly higher than that in the control group(P=0.005),and the mass of retroperitoneal(P=0.007),parametral(P=0.006)and inguinal(P=0.018)fat depot,serum triglyceride(P=0.009)and total cholesterol(P=0.000)were higher than those in the control group.Conclusion:The combination of DHT and high-fat diet can induce polycystic ovarian changes,estrous cycle changes and abnormal glucose and lipid metabolism in mice,which provides a mouse model for the experimental study of the pathogenesis of PCOS.