摘要
目的探讨多囊卵巢综合征患者卵巢颗粒细胞特异性表达基因甲基化水平.方法选取行体外受体-胚胎移植助孕治疗的女性患者颗粒细胞120例,分为多囊卵巢综合征组(因多囊卵巢综合征行体外受体-胚胎移植治疗)、对照组(因输卵管因素或男方因素行体外受体-胚胎移植治疗),每组60例.在行体外受体-胚胎移植治疗前卵泡期抽取外周静脉血,测定卵泡刺激素、黄体生成素、雌二醇、睾酮水平.采卵后收集卵泡液,测定雌二醇、雄烯二酮、抗苗勒氏管激素(Amh)水平.RT-PCR和Western blot检测细胞色素P45019A1(Cyp19a1)、Amh、抗苗勒氏管激素Ⅱ型受体(Amhr2) mRNA和蛋白表达水平.结果多囊卵巢综合征组卵泡刺激素、雌二醇/睾酮明显低于对照组,黄体生成素、雌二醇、睾酮、黄体生成素/卵泡刺激素明显高于对照组,差异具有统计学意义(P<0.05);卵泡液中雄烯二酮、Amh浓度明显高于对照组,雌二醇/雄烯二酮比值明显低于对照组,差异具有统计学意义(P<0.05).多囊卵巢综合征组卵巢颗粒细胞中Cyp19a1启动子甲基化水平明显高于对照组,Amh,Amhr2启动子甲基化水平明显低于对照组,差异具有统计学意义(P<0.05).Cyp19a1 mRNA和蛋白表达水平明显低于对照组,Amh,Amhr2 mRNA和蛋白表达水平明显高于对照组,差异具有统计学意义(P <0. 05). Pearson分析显示:Cyp19a1,Amh,Amhr2启动子的甲基化与mRNA表达呈负相关;Cyp19a1基因启动子的甲基化与卵泡液中雌二醇/雄烯二酮比值呈负相关.结论多囊卵巢综合征患者颗粒细胞中存在Cyp19a1,Amh,Amhr2基因甲基化,且与对应分子mRNA表达变化相关;Cyp19a1甲基化水平与卵泡液中雌二醇/雄烯二酮比值相关,提示Cyp19a1基因甲基化下调了芳香化酶活性,提升了卵巢雄激素合成水平.
Objective To investigate methylation level of ovarian granulosa cell-specific gene expression in patients with polycystic ovary syndrome( PCOS). Method 120 cases of granulosa cells from female patients undergoing in vitro recipient-embryo transfer were divided into polycystic ovary syndrome group( recipientembryo transfer in vitro for polycystic ovary syndrome) and control group( recipient-embryo transfer in vitro for fallopian tube or male factors),60 cases in each group. Peripheral venous blood was collected before in vitro recipient-embryo transfer in follicular phase. The levels of follicle stimulating hormone,luteinizing hormone,estradiol and testosterone were measured. Follicular fluid was collected after egg collection and the levels of estradiol,androstenedione and anti-Mullerian hormone( Amh) were determined.RT-PCR and Western blot were used to detect expression of cytochrome P45019 A1( Cyp19 a1),Amh and Amhr2. Results The levels of FSH,estradiol/testosterone in PCOS group were significantly lower than those in control group,luteinizing hormone,estradiol,testosterone and luteinizing hormone/FSH in PCOS group were significantly higher than those in control group( P< 0. 05),androstenedione and Amh in follicular fluid were significantly higher than those in control group,estradiol/androne.The diketone ratio was significantly lower than that of the control group( P<0.05).The methylation level of Cyp19 a1 promoter in ovarian granulosa cells of polycystic ovary syndrome group was significantly higher than that of control group,and the methylation level of Amh and Amhr2 promoter was significantly lower than that of control group( P<0.05).The expression of Cyp19 a1 was significantly lower than that of the control group,while the expression of Amh and Amhr2 was significantly higher than that of the control group( P < 0. 05). Pearson analysis showed that methylation of Cyp19 a1,Amh and Amhr2 promoters was negatively correlated with the expression of mRNA,and methylation of Cyp19 a1 promoter was negatively correlat
作者
郁晓冬
李红
Yu Xiaodong;Li Hong(Suzhou Municipal Hospital of Nanjing Medical University,Suzhou 215000,China)
出处
《北华大学学报（自然科学版）》
CAS
2019年第6期750-754,共5页
Journal of Beihua University（Natural Science）
基金
国家自然科学基金青年基金项目(81700913)
江苏省自然科学基金项目(BK2017240)
江苏省“强卫工程”医学重点学科基金项目(ZDXKB2016019)
关键词
多囊卵巢综合征
卵巢颗粒细胞
甲基化
polycystic ovary syndrome
ovarian granulosa cells
methylation
