【作者】 杨宗利；

【导师】 陈子江；

【作者基本信息】 山东大学 ， 妇产科学， 2009， 博士

【摘要】 目的多囊卵巢综合征(PCOS)是育龄期妇女常见的内分泌紊乱性疾病,胰岛素抵抗(IR)和高胰岛素血症与PCOS发病密切相关,它的家族聚集现象提示遗传因素在PCOS发病机制中起重要作用。尾加压素Ⅱ是一种血管活性肽,在胰岛素抵抗中发挥一定作用。本研究的目的是通过分析尾加压素Ⅱ(UTS2)基因rs228648(A/G)、rs2890565(A/G)的多态性在PCOS患者和正常对照之间基因型和等位基因频率的差异,及PCOS患者杂合子父母不同等位基因有无优势传递,比较不同基因型PCOS病人临床、内分泌及代谢特征的差别,研究相关基因多态性与PCOS不同表型的相关性,以探讨UTS2基因在PCOS发病及病理变化过程中的相关作用。方法本研究依据2003年鹿特丹标准选择PCOS患者101例及其父母202例,并选择同期就诊的105名健康妇女作为对照组。测定PCOS组及对照组基础状态下卵泡刺激素(FSH)、黄体生成素(LH)、睾酮(T)、空腹血糖(FPG)、胰岛素水平(FINS),测量身高、体重、腰围和臀围,计算体重指数(BMI)、腰臀比例(WHR)及胰岛素抵抗指数(HOMA-IR)。提取其抗凝血的DNA,根据所研究基因的序列设计相应引物,应用PCR扩增相应片段后,用熔解温度不同的基因分型法检测PCOS患者及其父母和对照组尾加压素Ⅱ基因rs228648(A/G)、rs2890565(A/G)的多态性并测序分析。结果(1)PCOS组与对照组相比,年龄无显著性差异,但PCOS组BMI,WHR高于对照组,LH和T水平也显著高于对照组,差异均有统计学意义;尾加压素Ⅱ基因rs228648A/G多态性在PCOS患者组及其父母组、对照组均符合Hardy-Weinberg平衡定律;PCOS组与对照组比较,基因型频率与等位基因频率均无明显差异;传递不平衡检验(TDT)显示,尾加压Ⅱ素基因rs228648A/G多态性位点杂合子父母2个不同等位基因无优势传递(P>0.05);尾加压素Ⅱ基因的SNPrs228648A/G多态性在PCOS-IR组及NIR组、肥胖组及非肥胖组的基因型分布及等位基因频率均无显著性差异(P>O.05),PCOS组尾加压素Ⅱ基因rs228648(A/G)不同基因型间BMI、WHR、LH、FSH、LH/FSH、TT、FPG、FINS、血脂差异无统计学意义(P>0.05),HOMA-IR在不同基因型间比较,差异有统计学意义(P<0.05)。GG基因型较携带A等位基因的PCOS患者HOMA-IR明显增高(2.33±2.06 vs1.83±1.08,P=0.038)。(2)尾加压素Ⅱ基因rs2890565A/G多态性在PCOS患者组及其父母组、对照组均符合Hardy-Weinberg平衡定律;SNPrs2890565在PCOS组AA基因型11例(10.89%),AG基因型43例(42.57%),GG基因型46例(46.53%);对照组AA基因型5例(4.76%),AG基因型34例(32.38%),GG基因型66例(62.86%),两组基因型频率比较差异有统计学意义(P<0.05)。PCOS组A等位基因频率(32.18%)明显高于对照组(20.95%),差异有统计学意义(P<0.05)。传递不平衡检验(TDT)显示,尾加压素Ⅱ基因rs2890565A/G多态性位点杂合子父母2个不同等位基因传递概率偏离50%(P<0.05),A等位基因优势传递;尾加压素Ⅱ基因的SNPrs2890565A/G多态性在PCOS-IR组AA+AG基因型频率63.6%,GG基因型36.4%,NIR组AA+AG基因型频率41.3%,GG基因型58.7%,IR组AA+AG基因型频率高于NIR组,两组比较差异有统计学意义(P<0.05)。PCOS-IR组A等位基因频率40.9%,G等位基因频率59.1%,NIR组A等位基因频率21.7%,G等位基因频率78.3%,IR组A等位基因频率显著高于NIR组,差异有统计学意义(P<0.01)。尾加压素Ⅱ基因的SNPrs2890565多态性在肥胖组及非肥胖组的基因型分布及等位基因频率无显著性差异(P>0.05),PCOS组尾加压素Ⅱ基因rs2890565(A/G)不同基因型间BMI、WHR、LH、FSH、LH/FSH、TT、血脂差异无统计学意义(P>0.05),FPG、FINS、HOMA-IR在不同基因型间差异有统计学意义。AA和AG基因型FPG较GG基因型明显增高(5.57±0.72 or 5.36±0.52 vs 4.02±0.59mmol/L,P<0.05),FINS较GG基因型明显增高(14.78±10.40 or11.24±6.13 vs 9.57±5.23 U/L,P<0.05)。AA基因型HOMA-IR较GG基因型明显增高(3.53±2.76 vs1.82±1.25 P<0.05)。结论(1)UTS2基因SNPrs228648A/G多态性与PCOS的临床特征,内分泌激素水平无相关性,但与胰岛素抵抗存在关联。(2)UTS2基因SNPrs2890565A/G多态性可能与PCOS发病相关,A等位基因与PCOS的胰岛素抵抗有关,与肥胖无密切关系。目的本研究的目的是通过分析脂联素受体1(ADIPOR1)基因rs1539355A/G多态性在PCOS患者和正常对照之间基因型和等位基因频率的差异,及PCOS患者杂合子父母不同等位基因有无优势传递,比较不同基因型PCOS病人临床、内分泌及代谢特征的差别,研究相关基因多态性与PCOS不同表型的相关性,以探讨ADIPOR1基因在PCOS发病及病理变化过程中的相关作用。方法本研究依据2003年鹿特丹标准选择PCOS患者101例及其父母202例,并选择同期就诊的105名健康妇女作为对照组。测定PCOS患者及对照组基础状态下卵泡刺激素(FSH)、黄体生成素(LH)、睾酮(T)、空腹血糖(FPG)、胰岛素水平(FINS),测量身高、体重、腰围和臀围,计算体重指数(BMI)、腰臀比例(WHR)及胰岛素抵抗指数(HOMA-IR)。提取其抗凝血的DNA,根据所研究基因的序列设计相应引物,应用PCR扩增相应片段后,用熔解温度不同的基因分型法检测PCOS患者及其父母和对照组脂联素受体1基因rs1539355A/G的多态性并测序分析。结果(1)脂联素受体1基因rs1539355A/G多态性在PCOS患者组及其父母组、对照组均符合Hardy-Weinberg平衡定律;PCOS组与对照组比较,基因型频率与等位基因频率均无明显差异,传递不平衡检验(TDT)显示,ADIPOR1基因rs1539355位点杂合子父母2个不同等位基因无优势传递(P>0.05);(2)ADIPOR1基因rs1539355A/G多态性在PCOS-IR组AA基因型为30.9%,AG+GG基因型69.1%;PCOS-NIR组AA基因型60.9%,AG+GG基因型39.1%,PCOS-IR组AG+GG基因型频率高于PCOS-NIR组,差异有统计学意义(P<0.05)。PCOS-IR组A等位基因频率61.8%,G等位基因频率38.2%;PCOS-NIR组A等位基因频率78.3%,G等位基因频率21.7%。PCOS组-IR组G等位基因频率高于PCOS-NIR组,差异有统计学意义(P<0.05)。ADIPOR1基因rs1539355A/G多态性在肥胖组与非肥胖组、高雄激素组与非高雄激素组基因型频率及等位基因频率无显著性差异(P>0.05)。(3)携带G等位基因的PCOS患者BMI、HOMA-IR较AA基因型明显升高(26.26±4.09 vs24.13±3.07 kg/m~2, 2.32±1.07 vs1.85±0.56,),差异有统计学意义(P<0.05),经一般线形模型控制BMI的影响后,携带G等位基因的PCOS患者HOMA-IR与AA基因型比较,差异仍有统计学意义(P=0.038)。结论(1)ADIPORI基因rs1539355A/G多态性可能与PCOS的胰岛素抵抗相关,并对BMI有一定影响。(2)ADIPOR1基因rs1539355A/G多态性与内分泌激素水平,脂代谢水平无相关性。目的探讨多囊卵巢综合征(PCOS)患者糖耐量受损(IGT)和糖尿病(NIDDM)的发生率及其高危因素。方法采用回顾性分析的方法,对101例PCOS患者行口服葡萄糖耐量(OGTT)实验后的临床资料进行分析,多因素logistic回归分析方法探讨PCOS患者糖耐量异常的危险因素。结果(1)根据葡萄糖耐量实验结果分成糖耐量异常组(AGT)24例(IGT22例、NIDDM2例)与糖耐量正常组(NGT)77例,IGT发生率21.8%,NIDDM发生率1.98%。(2)AGT组与NGT组相比,年龄、腰臀比(WHR)、体重指数(BMI)、睾酮、空腹血糖(FPG)、2小时血糖增高,差异有统计学意义(P<0.05),空腹胰岛素(FINS)、2小时胰岛素、稳态模式胰岛素抵抗指数(HOMA-IR)升高,差异具有统计学意义(P<0.01)。初潮年龄、黄体生成素/卵泡刺激素(LH/FSH)两组比较,差异无统计学意义(P>0.05)。(3)糖耐量异常组一级亲属有糖尿病家族史者15例,占62.5%(15/24),糖耐量正常组一级亲属有糖尿病家族史者24例,占31.2%(24/77),两组比较,糖耐量异常组一级亲属有糖尿病家族史者发生率明显增高,差异有统计学意义(P<0.01)。(4)多因素logistic回归分析显示,年龄、BMI、2型糖尿病家族史、空腹胰岛素为PCOS糖耐量异常的高危因素。结论多囊卵巢综合征发生糖耐量受损、糖尿病的危险性增加,葡萄糖耐量2小时血糖水平是PCOS糖耐量异常的较好监测指标。年龄、BMI、2型糖尿病家族史、空腹胰岛素是PCOS糖耐量异常发生的危险因素。更多还原

【Abstract】 Objective Polycystic ovary syndrome (PCOS) is one of the most common endocrine diseases of women of fertile age. Insulin resistance (IR) has a crucial role in the pathogenesis of this disorder. There is evidence for a genetic component in PCOS based on familial clustering of cases . UrotensinⅡhas been found to be a potent vasoactive peptide in humans and may be implicated in the development of insulin resistance. The objective of this study was to investigate the distribution of the single nucleotide polymorphisms of rs228648 and rs2890565 in urotension 2(UTS2) gene in PCOS patients and controls in a Chinese population , to reveal biased transmission of two different allels from heterozygous parents to affected daughters,to analyze the association of two single nucleotide polymorphisms with the clinical, the hormone and metabolic characteristics of PCOS, and to evaluate genetic influence of the two SNPs of UTS2 gene on the development of PCOS.Methods 101 Chinese Han families trios consisting of fathers , mothers, affected daughters with PCOS are recruited .The affected patients with PCOS were recruited according to the revised diagnostic criteria, announced in the 2003 American Society for Reproductive Medicine/European Society for Human Reproduction and Embryology (ASRM/ESHRE) Roterdam consensus. 105 women were selected as healthy controls. The physiological and biochemical parameters including height,weight, abdominal and hip circumference, levels of serum follicular stimulating hormone (FSH), luteinizing hormone (LH), testosterone, fasting glucose and insulin are measured. DNA was extracted from human leukocyte nuclei isolated from whole blood. The genotypes of rs228648 and rs2890565 in urotensionⅡgene were determined through polymerase chain reaction Tm-shift genotyping method and were further confirmed by DNA sequence analyses.Results (1)There is no significant difference in age between the PCOS patient and the control groups. The levelof T ,LH,BMI,and WHR is significantly higher in the PCOS group than that in control group.The distribution of genotypic frequency of SNP rs228648 in controls, PCOS group and their parents group did not deviate from the Hardy-Weinberg equilibrium; The distribution of three genotype and two allelotypes of SNP rs228648 in urotension gene were similar between PCOS group and control group. Transmission disequilibrium test ( TDT)did not show significantly biased transmission of two different allels from parents to affected daughters at rs228648 locus( P > 0.05); No significant differences between the distributions of the SNP rs228648 polymorphisms were observed when PCOS-IR group and PCOS-NIR group were compared The same results were found in obese PCOS women compared to non-obese PCOS women. There was no significant difference of clinical, hormone,lipids metabolism characteristics of different genenotype of the SNP rs228648 in PCOS women , but the carriers with GG genotype of SNP rs228648(A/G) had significantly higher HOMA-IR compared to those with AA and AG genotypes( 2.33±2.06 vsl.83±1.08,P=0.038).(2) The distribution of genotypic frequency of SNP rs2890565 in PCOS group and their parents group and control group did not deviate from the Hardy-Weinberg equilibrium; The frequency of AA,AG,GG genotype of SNP rs2890565 in PCOS group was 10.89%, 42.57%,46.53% respectively, while in control group was4.76%, 32.38%,62.86% respectively. The distribution of genotype of SNP rs2890565 in urotension gene differed significantly between PCOS group and control group (P<0.05) .The frequency of A allele in PCOS group and control group was 32.18%,20.95%, while G allele was 67.82%,79.05%. The frequency of A allele of SNP rs2890565 in urotension gene was significantly higher in PCOS group than in control group (P<0.05) .TDT analysis showed an excess of A allele from heterozygous parents to affected offspring (P<0.05). The frequency of AA+AG genotype of SNP rs2890565 in PCOS-IR group and PCOS-NIR group was 63.6%, 41.3% respectively, while GG genotype was36.4%,58.7% respectively. The distribution of genotype of SNP rs2890565 in urotension gene differed significantly between PCOS-IR group and PCOS-NIR group (P<0.05) .The frequency of A allele in PCOS-IR group and PCOS-NIR group was 40.9%,21.7%, while G allele was 59.1%,78.3%. The distribution of two alleles of SNP rs2890565 in urotension gene was different between PCOS-IR group and PCOS-NIR group (P<0.05) . There was no significant difference of hormone,lipids metabolism characteristics of different genenotype of the SNP rs2890565 in PCOS women ,but the carriers with AA or AG genotype of SNP rs2890565(A/G) had significantly higher fasting plasma glucose(5.57±0.72 or 5.36±0.52 vs 4.02±0.59mmol/L, PO.05), higner fasting insulin(14.78±10.40 or 11.24±6.13 vs 9.57±5.23 U/L, P<0.05 ) compared to those with GG genotypes . HOMA-IR with AA genotype was significantly higher than those with GG genotype (3.53±2.76 vsl.82±1.25 P<0.05) .Conclusions There was no association of SNP rs228648 with clinical,hormone characteristics in PCOS women, but was found to be associated with insulin resistance in PCOS women. The genetic polymorphism of urotensionⅡgene rs2890565(A/G) may be associated with PCOS , the higher frequency of A allele was associated with insulin resistance in PCOS but had no association with obesity. Objective The objective of this study was to investigate the distribution of the single nucleotide polymorphism of rsl539355 in adiponectin receptor 1(ADIPOR1) gene in PCOS patients and controls in a Chinese population , to reveal biased transmission of two different allels from parents to affected daughters,to analysis the association of the single nucleotide polymorphism with the clinical, the hormone and metabolic characteristics of PCOS, and to evaluate genetic influence of the SNP of ADIPOR1gene on the development of PCOS.Methods 101 Chinese Han families trios consisting of fathers , mothers, affected daughters with PCOS were recruited .The affected patients with PCOS were recruited according to the revised diagnostic criteria, announced in the 2003 American Society for Reproductive Medicine/European Society for Human Reproduction and Embryology (ASRM/ESHRE) Roterdam consensus. 105 women were selected as healthy controls .The physiological and biochemical parameters including height,weight, abdominal and hip circumference, levels of serum follicular stimulating hormone (FSH), luteinizing hormone (LH), testosterone, fasting glucose and insulin were measured. DNA was extracted from human leukocyte nuclei isolated from whole blood. The genotypes of rs1539355 locus in ADIPOR1 gene were determined through polymerase chain reaction Tm-shift genotyping method and were further confirmed by DNA sequence analyses.Results There was no significant difference in age between the PCOS patients and the control group . The levelof T ,LH,BMI,and WHR was significantly higher in the PCOS group than that in control group.The distribution of genotypic frequency of SNP rs1539355 in controls, PCOS group and their parents group did not deviate from the Hardy-Weinberg equilibrium; The distribution of three genotype and two allelotypes of SNP rs1539355 inADIPOR1 gene were similar between PCOS group and control group. Transmission disequilibrium test ( TDT) did not show significantly biased transmission of two different allels from parents to affected daughters at rs1539355 locus( P > 0.05); (2) The frequency of AG+GG genotype of SNP rsl539355 in PCOS-IR group and PCOS-NIR group was 69.1%, 39.1% respectively, while AA genotype was30.9%,60.9% respectively. The distribution of genotype of SNP rsl539355 in urotension gene differed significantly between PCOS-IR group and PCOS-NIR group(P<0.05) .The frequency of A allele in PCOS-IR group and PCOS-NIR group was 61.8%,78.3%, while G allele was 38.2%,21.7%. The distribution of two alleles of SNP rs1539355 in ADIPOR1 gene were different between PCOS-IR group and PCOS-NIR group (P<0. 05) . The distribution of frequency of genotypes and alleles was similar in hyperandrogenism group and non- hyperandrogenism group,obese group and non-obese group (3) The carriers with G allele had higher body mass index (BMI) ( P<0. 05) and HOMA-IR ( P <0. 05) compared to AA genotype in the patients with PCOS.(26. 26±4.09 vs24. 13±3. 07 kg/m~2, 2.32±1.07 vs1. 85±0. 56) . A univariate general linear model analysis, introducing BMI as a covariate in the model, was perfomed , the results showed that SNP rsl539355 still influenced HOMA-IR( P <0.05).Conclusions SNP rs1539355 in ADIPOR1 gene was found to be associated with insulin resistance and had some influence on BMI, but there was no association with hormonal and lipid characteristics in PCOS women. Objective To analyse the prevalence of impaired glucose tolerance (IGT) and diabetes mellitus(NIDDM) in women with polycystic ovary syndrome and to identify associated risk factors within this population.Method Retrospective study was performed in 101 women with PCOS and clinical data were analysed after oral glucose tolerance tests to identify risk factors for abnormal glucose tolerance by using multivariate logistic regression.Results Based on the criteria for IGT and NIDDM, PCOS women were divided into two groups :77 cases with normal glucose tolerancr(NGT) and 24 cases with abnormal glucose tolerance. The prevalence for IGT was 21.8% and that of NIDDM was 1.98%. The mean age, body mass index(BMI), waist hip ratio(WHR), total testosterone, fasting plasma glucose, 2-h plasma glucose level were significantly different between AGT and NGT group(P<0.05). Fasting insulin, 2-h insulin, homeostasis model assessment of insulin resistance were significantly higher in AGT group(P＜0.01). Menarche age and LH/FSH made no difference between the two groups(P>0.05). The incidence of a family history of diabetes was higher in AGT patients(P<0.01). Multivariate logistic regression showed that age, BMI, , a family history of diabetes,fasting insulin levels were independent risk factors for AGT.Conclusion Women with PCOS are at increased risks for abnormal glucose metabolism. 2-h plasma glucose level after oral glucose toerance test was a significant predictor for abnormal glucose tolerance in women with PCOS. Age, BMI, a family history of diabetes, fasting insulin levels were independent risk factors for abnormal glucose tolerance.更多还原