高雄激素PCOS动物模型中抗苗勒管激素Ⅱ型受体蛋白及其基因甲基化分析  

Expression of anti-müllerian hormone type Ⅱ receptor and its gene methylation in rat model of PCOS

作　　者：郝栋栋 殷倩[1] 钟兴明[2] 韦相才[1,2] 苗竹林[2] 王永霞[2] 

机构地区：[1]暨南大学附属第一医院妇产科,广州510632 [2]广东省计划生育科学技术研究所,广州510000

出　　处：《生殖医学杂志》2016年第9期832-837,共6页Journal of Reproductive Medicine

基　　金：广东省科技计划项目(2009B030801270);广东省自然科学基金项目(10151008010000000)

摘　　要：目的通过建立高雄激素诱导的多囊卵巢综合征(PCOS)大鼠模型,探讨抗苗勒管激素Ⅱ型受体(AMHRⅡ)在卵巢、子宫内膜中的表达及其基因启动子区域甲基化与PCOS发病及局部病理变化的关系。方法 45只雌性SD大鼠随机分两组:PCOS组皮下注射脱氢表雄酮(DHEA)建立PCOS模型,对照组同期皮下注射等量生理盐水;测定血清激素水平,结合卵巢组织HE染色切片结构改变评估PCOS模型;免疫组化法检测卵巢、子宫内膜组织AMHRⅡ蛋白的表达与定位;通过甲基化特异性PCR(MSP)定性分析两组大鼠血液、卵巢组织AMHRⅡ基因启动子区域的甲基化状态。结果血清性激素水平及卵巢切片结果均提示成功建立PCOS动物模型;免疫组化检测显示AMHRⅡ蛋白定位表达于细胞膜,在卵巢间质及卵泡颗粒细胞中均存在阳性表达,且在模型组中的表达强于对照组(P<0.05);AMHRⅡ在子宫内膜中也存在表达,两组之间表达无统计学差异(P>0.05);MSP结果显示两组中血液及卵巢组织所有DNA样本均检出部分甲基化条带,两组之间无统计学差异(P>0.05)。结论 AMHRⅡ可能参与了PCOS中AMH调控卵泡发育的过程,但与子宫内膜病变无明显相关性;AMHRⅡ基因在启动子区域存在着部分甲基化,但这种甲基化状态与PCOS发病及局部组织病理变化无明显相关。Objective： To study localization, distribution and expression of anti-mullerian hormone type Ⅱ receptor （AMHR Ⅱ ） in ovary and endometrium based on rat model of polycystic ovary syndrome （PCOS）,and explore the correlation between AMHR Ⅱ gene methylation and the local histopathological changes. Methods： Forty five female Sprague Dawley rats were grouped randomly into model group （PCOS model） subcutaneously injected with dehydroepiandrosterone （DHEA） and control group with sodium chloride instead. The levels of related reproductive hormones in serum were measured. The ovarian tissue of rats in every group underwent paraffin embedding, microsection, HE staining, and then patho-histological changes were observed to assess the establishment of PCOS model. The localization, distribution and semi quantitative analysis of AMHR Ⅱ protein were determined by immunohistochemistry （IHC）. The qualitative analysis of AMHR Ⅱ gene methylation in blood and ovarian tissues was performed by methylation-specific PCR （MS-PCR）. Results： The reproductive hormone levels and morphology analysis of tissue HE stained indicated that PCOS animal model was successfully established. IHC analysis showed that AMHR Ⅱ mainly located and expressed in cell membrane, and ovarian stroma and of granulosa cells of follicle presented positively expression. The expression was significantly higher in the model group than the control group （P〈0.05）. AMHR Ⅱwere also expressed in endometrial glandular epithelial cells and stromal cells, and there were no significant differences between the two groups （P 〈 0.05）. MSP qualitative analysis showed partial methylation occurred in the promoter region of AMHR Ⅱgenes in the blood and ovarian tissues in both groups,but without significant differences （P〈0.05）. Conclusions： AMHR Ⅱ may be involved in the follicular development process regulated by AMH, but it may not be related to endometrial lesions. AMHR Ⅱ gene partial methylation in the promoter

关 键 词：抗苗勒管激素Ⅱ型受体 多囊卵巢综合征 DNA甲基化 动物模型 

分 类 号：R[医药卫生]