中国汉族 CYP11a微卫星多态性与多囊卵巢综合征高雄性激素血症关系的研究

目的探讨胆固醇侧链裂解酶CYP11α基因微卫星多态性与多囊卵巢综合征(polycystic ovary syndrome，PCOS)患者高雄激素血症形成的关系。方法应用聚合酶链反应及聚丙烯酰胺凝胶电泳分离技术检测86例PCOS患者及50名对照妇女CYP11α基因微卫星多态性及其分布，比较CYP11α(tttta)n各等位基因与PCOS高雄激素之间的关系。结果CYP11α基因有4种等位基因(4、6、8、9)；PCOS及对照组基因频率分布为0．17，0．31，0．39，0．13和0．22，0．35，0．33，0．10，差异无统计学意义；CYP11α(tttta)n各等位基因与PCOS高雄激素无明显关系。结论中国汉族CYP11α微卫星多态性对PCOS高雄激素血症的形成无明显关系，不是PCOS的主要致病因素。     

Association study of CYP17 and HSD11B1 in polycystic ovary syndrome utilizing comprehensive gene coverage

Cytochrome P450-C17 enzyme (CYP17) is an important component of the androgen synthesis pathway, a pathway that is dysfunctional in polycystic ovary syndrome (PCOS). Variation in 11-beta hydroxysteroid dehydrogenase (HSD11B1) is associated with cortisone reductase deficiency, a condition with a phenotype similar to PCOS. Both CYP17 and HSD11B1 genes have been previously studied for their possible relationship with PCOS, yielding inconsistent results. In this study, we evaluated the association between variation in these genes and PCOS. Two-hundred and eighty-seven Caucasian PCOS women and 187 Caucasian controls were genotyped for single nucleotide polymorphisms (SNPs) that were specifically chosen to allow full coverage of CYP17 and HSD11B1, including four SNPs in CYP17 and eight SNPs in HSD11B1. SNP and haplotype association analyses were conducted. Our results indicate that variants in the two genes are not associated with PCOS, or with the quantitative traits characteristic of PCOS, suggesting that these genes are not major risk factors for the syndrome.     

CYP11α基因微卫星多态性与多囊卵巢综合征关系的研究

目的探讨胆固醇侧链裂解酶CYP11α基因启动子中（TTTTA）n多态性和多囊卵巢综合征的关系。方法测定63例PCOS患者和40例正常妇女血清中的性激素水平,同时检测CYP11α基因微卫星多态性及其分布。结果 CYP11α基因在受检人群中存在5种等位基因;各基因型在PCOS患者与正常人之间差异无统计学意义;PCOS患者主要基因型之间血清性激素水平也无统计学差异。结沦CYP11α基因微卫星多态性与PCOS的发病之间无直接相关性,不是P-COS的主要致病因素。     

A common polymorphism in the human aromatase gene alters the risk for polycystic ovary syndrome and modifies aromatase activity in vitro

Aromatase is a key enzyme involved in estradiol and estrone biosynthesis. Given that polymorphisms of the CYP19A1 gene encoding aromatase have been correlated with plasma testosterone levels, CYP19A1 may therefore act as a genetic modifier of the hyperandrogenic phenotype of polycystic ovary syndrome (PCOS). However, no functional CYP19A1 polymorphisms that predict the risk of PCOS have been identified. We explored the role of CYP19A1 genetic variation in a large case-control study involving 1078 samples, in which five common genetic polymorphisms were scored. Human embryonic kidney 293 cells were transiently transfected with a vector encoding either the CYP19A1 wild-type (WT) allele or an Arg(264)Cys variant to evaluate aromatase activity. Cells were cultured with androstenedione and estrone levels were measured using a specific ELISA. The Arg(264)Cys variant of CYP19A1 (rs700519) is associated with PCOS (P= 0.004, corrected P = 0.02). In this functional study, when cells were cultured in varying concentrations of androstenedione (100, 400 and 500 nM), transfection with the Arg(264)Cys variant resulted in increased conversion of androstenedione to estrogen when compared with transfection with the WT construct (P< 0.001). Our data suggest that the common missense polymorphism rs710059 is associated with susceptibility to PCOS and that the Arg(264)Cys variant may increase aromatase enzymatic activity. Overall, these findings imply that aromatase plays an important role in PCOS.     

来曲唑诱导大鼠多囊卵巢综合征模型卵巢组织中CYP19基因的表达

目的:研究来曲唑诱导的多囊卵巢综合征(PCOS)模型大鼠卵巢组织中CYP19基因的表达,为该模型的应用提供一定的理论依据.方法:SD大鼠随机分为模型组和对照组,模型组应用来曲唑诱导大鼠PCOS模型.放射免疫法测定大鼠血清性激素水平;H-E染色观察卵巢组织学变化,免疫组织化学、Real time-PCR和免疫印迹法检测CYP19基因在卵巢组织中的表达.结果:模型组血清性激素水平睾酮、黄体生成素、卵泡刺激素浓度显著增高,雌二醇、孕酮浓度降低.模型组CYP19mRNA及蛋白表达均显著高于对照组.结论:来曲唑诱导的PCOS大鼠卵巢组织中CYP19基因表达水平增高.     

Candidate genes in polycystic ovary syndrome

The candidate gene approach has already paid some dividends in trying to understand the complex genetics of polycystic ovary syndrome (PCOS). In terms of steroidogenic abnormalities, CYP11a-encoding P450 side chain cleavage-appears to be a major susceptibility locus. In relation to the well-described metabolic disturbances in PCOS, the insulin gene variable number tandem repeat (VNTR) appears to be a promising candidate, at least in populations studied in the UK. Finally, genes implicated in ovarian follicular development may have a role in the aetiology of PCOS, as demonstrated by recent identification of the follistatin gene as a potential disease locus. It seems unlikely that PCOS can be explained on the basis of a single gene disorder although, in a given family, one gene may have a predominant effect. An oligogenic model seems the most appropriate basis on which to understand the genetic origins of this very common disorder. The candidate gene approach has been useful to date, but it may prove important in the near future to perform an anonymous genome-wide scan to identify hitherto unheralded susceptibility loci.     

Association study for single nucleotide polymorphisms in the CYP17A1 gene and polycystic ovary syndrome

Women with polycystic ovary syndrome (PCOS) are characterized by excess androgen secretion and anovulatory infertility as a cause of follicular maturation arrest, and they are also associated with insulin resistance and obesity. Recently, it was suggested that one of the etiologies for PCOS is an abnormality of steroid hormones, and excessive secretion of androgen. The endoplasmic reticular cytochrome P450, 17alpha-hydroxylase (CYP17A), plays a key role in the mechanism of steroid hormones such as adrenal and gonadal steroid biosynthesis. Therefore, we studied the association between single nucleotide polymorphisms (SNPs) of the A1 allelic variant of the CYP17 gene and PCOS in a Korean population. The study recruited 134 Korean women with PCOS and 100 healthy women as controls. Using the HapAnalyzer, the genotype of the CYP17A1 polymorphism in PCOS and control patients were analyzed. We considered a p-value lower than 0.05 to be statistically significant. After genotypic analysis, we found seven SNPs of the CYP17A1 gene in a large population of subjects. The frequency of seven SNPs had no significant association with PCOS. However, one haplotype (ht3) had a p-value of p=0.001, suggesting that it may be associated with the pathogenesis of PCOS in a Korean population.     

The genetic basis of polycystic ovary syndrome

Polycystic ovary syndrome (PCOS) is the most common endocrinopathy in women of reproductive age. Familial clustering of cases suggests that genetic factors play an important part in its aetiology. A number of studies of families with several cases of PCOS have produced results suggesting an autosomal dominant trait. Detailed analysis of a large number of affected families has, however, cast some doubt about the mode of inheritance. An autosomal dominant trait remains possible but a more complex aetiology seems more likely. The results of our recent studies support the concept of an oligogenic disorder in which genes affecting metabolic pathways in glucose homeostasis and steroid biosynthesis are both involved. We review evidence for an important role for the insulin gene minisatellite in the aetiology of anovulatory PCOS and for the gene coding for P450 cholesterol side chain cleavage (CYP11a) in the mechanism of excessive androgen secretion in women with polycystic ovaries. We propose that the heterogeneity of clinical and biochemical features in PCOS can be explained by the interaction of a small number of key genes with environmental, particularly nutritional, factors.      

No evidence of mutations in the P450 aromatase gene in patients with polycystic ovary syndrome

BACKGROUND: Etiology and inheritance pattern in polycystic ovary syndrome (PCOS) remain uncertain. Granulosa cells from follicles of women with PCOS have little, if any, aromatase (encoded by the CYP19 gene) activity; follicles contain low levels of estradiol, P450arom mRNA and aromatase stimulating bioactivity. Mice with targeted disruption of the CYP19 gene present cystic follicles. It has been proposed that chronic exposure to high levels of LH, because of aromatase deficiency, determines the development of ovarian cysts. Herein, we investigated if mutations in the CYP19 gene and/or its ovary promoter are causal in patients with PCOS. METHODS: Twenty-five patients with PCOS and 50 control women were studied. PCR analysis of genomic DNA and complete sequence of all exons of the aromatase gene and its ovary promoter were performed. RESULTS: No heterozygous or homozygous mutant alleles were present in any of the patients studied. CONCLUSIONS: In the population studied, mutations of the P450arom gene or its promoter are not the cause of PCOS. However, these findings do not preclude the possible importance of an aromatase disorder in PCOS etiology. Variations in aromatase complex function could play a role in PCOS etiology, but the determinants of such variations might be located in other genes.     

Polymorphism in the peroxisome proliferator-activated receptor-gamma gene in women with polycystic ovary syndrome

BACKGROUND: In view of the strong evidence implicating peroxisome proliferator-activated receptor-gamma (PPARgamma) in adiposity and insulin resistance a study was carried out to investigate PPARgamma genotype frequencies in women with polycystic ovary syndrome (PCOS) and to elucidate its role in the pathogenesis of the syndrome. METHODS: The study involved 135 women with PCOS and 115 healthy control women who were genotyped for a known functional variant of the PPARgamma gene using single strand conformation polymorphism (SSCP) analysis. RESULTS: A significantly different allele distribution of the Pro12 Ala polymorphism of the PPARgamma gene was observed between the two groups, with the frequency of the variant Ala isoform being significantly reduced in the PCOS group (12.6%) when compared with the control group (19.1%) (P = 0.045), at an odds ratio of 0.609 (95% confidence interval: 0.374-0.991). The genotype distributions of the Pro12 Ala polymorphism in the PCOS and control groups were different with borderline significance (P = 0.051). CONCLUSIONS: Our data support a role for PPARgamma gene polymorphism in the pathogenesis of PCOS, the presence of the Ala isoform being protective against the development of PCOS.     

Association of polycystic ovarian syndrome with human leukocyte antigen polymorphism in Korean women

Although several studies have demonstrated the genetic contribution to polycystic ovarian syndrome (PCOS), the cause of this syndrome remains unclear. The aim of this study was to elucidate the relationship between human leukocyte antigen (HLA) systems and PCOS in Koreans. We compared the HLA-A, B and DRB1 genotype distribution of 52 PCOS patients and 67 healthy Korean women. In addition, we investigated the association of HLA with free-testosterone level. HLA-A*11, A*31 and B*54 showed increased phenotype frequencies (PFs) in PCOS women compared to controls (p = 0.032, OR 2.79; p = 0.019, OR 6.05; p = 0.002, OR 6.40). HLA-DRB1*15 showed negative correlations with the free-testosterone concentration both in total subjects and PCOS patients (p = 0.024 and p = 0.008). The results of the study suggest mild associations of HLA alleles with pathophysiology of PCOS and/or testosterone production in PCOS. Further investigation in a large number of subjects, including subdivision and multi-population studies, will need to be conducted to prove the consistent or variable association in PCOS.     

The association between polycystic ovaries and endometrial cancer

BACKGROUND: Women with polycystic ovary syndrome (PCOS) are assumed to be at increased risk of endometrial cancer (EC), albeit of a more differentiated type with better prognosis than in normal women. This study was designed to test these assumptions, as evidence for them is lacking. METHODS: The prevalence of polycystic ovaries (PCO), as a marker of PCOS, was investigated in ovarian sections from 128 women with EC and 83 with benign gynaecological conditions. The expression of the prognostic markers p53, Ki67, Bcl2 and cyclin D1 was also investigated by immunohistochemistry in endometrial tumours from 11 women with PCO and 16 with normal ovaries. RESULTS: Overall, PCO were similarly prevalent in women with EC (8.6%) and benign controls (8.4%); however, in women aged <50 years, PCO were more prevalent in women with EC (62.5 versus 27.3%, P = 0.033). Cyclin D1-expressing endometrial tumours tended to be more prevalent in women with PCO compared to normal ovaries (36.4 versus 6.25%, respectively, P = 0.071). Bcl2-, p53- and Ki67-expressing tumours were similarly prevalent. CONCLUSIONS: The association between PCOS and EC appears confined to premenopausal women. The tendency for cyclin D1-expressing endometrial tumours to be more prevalent in women with PCO challenges the assumption that EC prognosis is improved in women with PCOS.     

Polycystic ovary syndrome in post-menopausal women--marker of the metabolic syndrome

OBJECTIVES: To determine among post-menopausal women, the prevalence of polycystic ovary syndrome (PCOS) and to evaluate the endocrine and metabolic profile of these women in comparisons to women without PCOS. METHODS: All women attending two post-menopausal outpatient clinics, fulfilling the inclusion criteria, were evaluated by physical examination, detailed questionnaire and laboratory measurements of glucose, lipids and sex-hormone levels. RESULTS: Among 104 women 7 (6.7%) were diagnosed with PCOS. As compared to women without PCOS, central obesity was more common in the PCOS group (p=0.004), 4/7 had type 2 diabetes compared to 8/97 in the non-PCOS group (p=0.003) and 6/7 versus 31/97 had the characteristic dyslipidemia of the metabolic syndrome (p=0.047). CONCLUSIONS: PCOS is common in post-menopausal women attending outpatient's clinics, and is a marker for a metabolic profile that is associated with a high-risk for cardiovascular diseases (CVD).     

The (TAAAA)n microsatellite polymorphism in the SHBG gene influences serum SHBG levels in women with polycystic ovary syndrome

BACKGROUND: Hyperandrogenaemia is a common feature of polycystic ovary syndrome (PCOS). The sex hormone-binding globulin (SHBG) gene was proposed as being a PCOS candidate gene. A possible influence of the microsatellite polymorphism (TAAAA)(n) in the SHBG gene on serum SHBG levels in PCOS patients was investigated. METHODS: One hundred and twenty-three PCOS patients and 110 age-matched controls were included in the study. Peripheral blood samples were obtained. Genotyping of the (TAAAA)(n) polymorphism in the SHBG gene was performed. Serum LH, FSH, SHBG and total testosterone concentrations were determined. RESULTS: SHBG alleles with 6-11 TAAAA repeats were found. None of the SHBG alleles or genotypes were present at a significantly more frequent rate in PCOS patients compared with controls. Serum SHBG levels were significantly lower (P < 0.001) in PCOS patients compared with controls and were found to be strongly influenced by the (TAAAA)(n) SHBG polymorphism, in both the PCOS (55.3%) and control (33.1%) groups of patients. CONCLUSIONS: The (TAAAA)(n) SHBG gene polymorphism might be an important predictor for serum SHBG levels and, consequently, for hyperandrogenaemic clinical presentation of PCOS.     

Evaluation of glucose metabolism and reproductive hormones in polycystic ovary syndrome on the basis of peroxisome proliferator-activated receptor (PPAR)-gamma2 Pro12Ala genotype

BACKGROUND: Peroxisome proliferator-activated receptor (PPAR)-gamma2 Pro12Ala polymorphism has been suggested as a protective factor for polycystic ovary syndrome (PCOS). In this study, we aimed to investigate metabolic features and reproductive hormones in women with PCOS and compare these features with control women on the basis of Pro12Ala genotype. METHODS: This study involved 60 randomly selected women with PCOS and 60 controls. Main outcome measures were anthropometric measures, variables of glucose metabolism and reproductive hormones. All the patients were genotyped for Pro12Ala variant of PPAR-gamma2 gene. RESULTS: Patients with Pro12Ala polymorphism were more obese in both groups. Furthermore, they had lower fasting insulin levels, were less insulin-resistant and were less glucose-intolerant as demonstrated by 2 h glucose concentrations. However, there was no difference in reproductive hormone levels on the basis of Pro12Ala genotype. CONCLUSIONS: Both control women and women with PCOS had significant differences in glucose metabolism on the basis of PPAR-gamma2 Pro12Ala polymorphism. Pro12Ala variant may break the process that leads to PCOS in susceptible women, instead of being a direct causal relationship between Pro12Ala polymorphism and PCOS.     

No association between the -308 polymorphism in the tumour necrosis factor alpha (TNFalpha) promoter region and polycystic ovaries

The tumour necrosis factor (TNF)2 allele appears to be linked with increased insulin resistance and obesity, conditions often found in overweight patients with polycystic ovary syndrome (PCOS). The significance of TNFalpha polymorphism in relation to the clinical and biochemical parameters associated with PCOS was investigated in 122 well-characterized patients with polycystic ovaries (PCO). Of these, 84 had an abnormal menstrual cycle and were classified as having PCOS, while the remaining 38 had a normal menstrual cycle and were classified as having PCO. There were a further 28 individuals without PCO (non-PCO) and 108 individuals whose PCO status was undetermined (reference population). The promoter region of the TNFalpha gene was amplified by polymerase chain reaction (PCR), and the presence or absence of the polymorphism at -308 was determined by single-strand conformational polymorphism (SSCP) analysis. The less common TNF allele (TNF2) was found as TNF1/2 or TNF2/2 in 11/38 (29%) of PCO subjects, 25/84 (30%) of PCOS subjects, 7/28 (25%) of non-PCO subjects, and 45/108 (42%) of the reference population. There was no significant difference in the incidence of the TNF2 allele between the groups. The relationship of TNF genotype to clinical and biochemical parameters was examined. In both the PCO group and the PCOS group, the presence of the TNF2 allele was significantly associated with lower glucose values obtained from the glucose tolerance testing (P<0.05). The TNF genotype was not significantly associated with any clinical or biochemical parameter measured in the PCO, PCOS or non-PCOS groups. Thus, the TNFalpha -308 polymorphism does not appear to strongly influence genetic susceptibility to polycystic ovaries.     

Depression in women with polycystic ovary syndrome: clinical and biochemical correlates

BACKGROUND: We assessed the prevalence of mood disturbance among women with prospectively documented polycystic ovary syndrome (PCOS). METHODS: Thirty-two women with PCOS completed the Center for Epidemiological Studies-Depression Rating Scale (CES-D). Clinical and biochemical characteristics were assessed. RESULTS: Sixteen women had CES-D scores indicative of depression. Depression was associated with greater insulin resistance (P=0.02) and higher body mass index (P=0.05). Women receiving oral contraceptives for the treatment of PCOS were less depressed than patients not receiving treatment (P=0.03). LIMITATIONS: Possible selection bias, use of a screening tool alone without further diagnostic evaluation of depression, small samples size and lack of direct comparison with an age matched control group, should be considered in interpretation of these results. CONCLUSION: Findings suggest a high prevalence of depression among women with PCOS, and an association between depression and PCOS markers.     

Relationship between E23K (an established type II diabetes-susceptibility variant within KCNJ11), polycystic ovary syndrome and androgen levels

Polycystic ovary syndrome (PCOS) is strongly associated with hyperinsulinaemia and type II diabetes (T2D). Sequence variation within KCNJ11 (encoding Kir6.2, the beta-cell inwardly rectifying potassium channel) is implicated in the pathogenesis of neonatal diabetes, hyperinsulinaemia of infancy and multifactorial T2D. Comprehensive tagging studies have demonstrated that the KCNJ11 E23K variant (or ABCC8 A1369S in LD>0.9) is responsible for the known association between KCNJ11 and T2D. Given the phenotypic overlap between PCOS and T2D, we investigated whether E23K is involved in susceptibility to PCOS and related traits. Case-control analyses for the KCNJ11 E23K variant were performed in (a) 374 PCOS cases and 2574 controls of UK British/Irish origin, and (b) 550 women with PCOS symptoms and 1114 controls from a Finnish birth cohort. The relationship between E23K genotype and androgen levels (a key intermediate phenotype relevant to PCOS) in 1380 samples was studied. The UK case-control analysis revealed no association between E23K genotypes and PCOS status (P=0.49; Cochran-Armitage test), and no significant relationship between E23K genotype and androgen measures in the samples for which these phenotypes were available (P=0.19). Similarly, the Finnish case-control analysis showed no association between E23K genotypes and PCOS status (P=0.75; Cochran-Armitage test), and no significant relationship between E23K genotype and androgen measures in the samples for which these phenotypes were available (Finnish controls, P=0.25; Finnish cases, P=0.08). In conclusion, these data (involving >4600 subjects) provide no evidence that common variants of the KCNJ11 E23K polymorphism have a major influence on PCOS susceptibility, though modest effect sizes (OR<1.25) cannot be excluded.     

The PON1-108C/T polymorphism, and not the polycystic ovary syndrome, is an important determinant of reduced serum paraoxonase activity in premenopausal women

BACKGROUND: Because serum paraoxonase activity is influenced by the -108C/T polymorphism in the PON1 gene, we studied its involvement in the decreased paraoxonase activity recently described in the polycystic ovary syndrome (PCOS). METHODS: Paraoxonase activity, PON1-108C/T genotypes and clinical, hormonal and biochemical variables were evaluated in a case-control study involving 139 consecutive PCOS patients and 85 healthy controls matched for BMI and prevalence of smoking. RESULTS: Women homozygous for -108T presented with reduced serum paraoxonase activity compared with carriers of C alleles (P < 0.001), both in PCOS patients and in controls. Although homozygosity for T alleles was more prevalent in PCOS patients than in controls (P = 0.003), serum paraoxonase activity was not significantly different in the PCOS and control groups. In a stepwise multivariate linear regression model, homozygosity for PON1-108T alleles was the only significant predictor of the logarithm of serum paraoxonase activity (beta = -0.328, t = -4.176, P < 0.001). CONCLUSIONS: In premenopausal women from the Spanish population, the PON1-108C/T polymorphism, and not PCOS, is an important determinant of serum paraoxonase activity.     

多囊卵巢综合症辅助诊断系统的开发

开发多囊卵巢综合症辅助诊断系统,帮助医生诊断多囊卵巢综合症患者的类型。根据大量多囊卵巢综合症患者症候和证型的数据,从中总结出对应关系,以期通过症候判断患者所属的证型。多囊卵巢综合症患者分类效果良好。其中Logistic算法的正确率为93.5%。设计出的多囊卵巢综合症辅助诊断系统科学合理、简单易用、具有实际指导意义。