胰岛素和人绒毛膜促性腺激素建立高雄激素多囊卵巢大鼠模型

A Rat Polycystic Ovary Model Established by Combined use of Insulin and Human Chorionic Gonadotrophin

目的:尝试通过联合注射胰岛素和人绒毛膜促性腺激素(hCG),尝试建立一种与多囊卵巢综合征(PCOS)发病机制相近,并有高雄激素分泌的大鼠动物模型。方法:将出生2个月的雌性SD大鼠分为实验组和对照组,第1～10天实验组大鼠注射胰岛素,剂量由0.5IU/d逐步增加至6.0IU/d,第11～22天注射固定剂量胰岛素6.0IU/d,并同时加用6.0IU/d的hCG分2次注射。第23天将大鼠处死,称重后取卵巢分析组织形态学,采集躯干血测定激素水平。应用RT-PCR和蛋白质印迹比较两组大鼠卵巢组织细胞CYP17基因的mRNA和蛋白表达。结果:用药结束与对照组相比,实验组大鼠体质量增加(53.9±5.6)g(P<0.05),卵巢呈多囊样变质量为(0.13±0.017)g(P<0.05),雄稀二酮和睾酮的水平升高分别为(08±0.16)μg/L和(4.98±0.5)μmol/L(P<0.05),17-羟孕酮和孕酮的水平也升高分别为(10.79±1.78)μg/L和(108.04±24.83)μg/L(P<0.05),脱氢表雄酮硫酸盐无差异(P>0.05)。实验组大鼠卵巢组织CYP17的RNA水平与对照组相比升高且蛋白表达也增加(P<0.05)。结论:联合使用胰岛素和hCG可建立与PCOS发病类似且CYP17表达异常增高的大鼠模型,该模型在研究PCOS高雄激素血症方面具有独特优势。

Objective： To establish a rat model of hyperandrogenism with pathogenesis similar to PCOS by combined administration of insulin and hCG. Methods：Two month old female SD rats were divided into experimental and control groups. From days 1 to 10, the experimental rats were injected with slowly increasing doses of insulin, from 0.5 IU/d to 6.0 IU/d. Between days 11 and 22, the insulin was injected at a fixed dose of 6.0 IU/d concurrently with hCG （6.0 IU/d） injected twice per day. Rats were sacrificed on day 23. Ovarian histology was examined, and trunk blood was collected for detection of hormone levels. RT-PCR and Western-blot analyses were used to compare the mRNA and protein expression of CYP17 in ovarian cells. Results： Compared with rats from the control group, the rats in the experimental group gained more weight [ （53.9±5.6） g, P〈0.05 ], the weight of the ovaries increased [ （0.13±0.017 ）g, P〈0.05 ], and the ovaries become polycystic. Androstenedione and testosterone levels were increased to [ （0.8±0.16）μg/L] and [ （4.98±0.5）μmol/L（P〈0.05） 1, respectively, and the levels of 17-hydroxyprogesterone and progesterone were also elevated to （ 10.79 ±1.78）μg/L and （ 108.04±24.83 ） μg/L （P〈0.05）. However, there were no significant differences in dehydroepiandrosterone sulfate levels between the two groups （P〉0.05）. Both the CYP17 mRNA and protein expression in the ovaries of rats from the experimental group were higher than in the control group （P〈0.05）. Conclusions： A rat model that capitulates the features of PCOS and has amplified CYP17 gene function, can be established by combined administration of insulin and hCG. This model may provide a unique advantage for research about hyperandrogenism and PCOS.