The Related Candidate Gene And Clinical Research Of Premature Ovarian Failure

Objective:To investigate the relevance between the androgen receptor (AR) gene CAG repeat polymorphism and Fragile X mental retardation gene-1(FMR1) gene CGG repeat polymorphism with the susceptibility of Premature Ovarian Failure (POF); To study the change of lipid metabolism and bone metabolism and the effect of different regimens of hormone therapy on their bone mass and lipid metabolism in women with POF and to find adaptive therapy for infertility women with POF.Method:85patients were recruited (2009June to2011June) from the gynecological endocrine outpatient clinic of Tianjin Medical University General Hospital and the reproductive medicine clinic of the first hospital of QinHuangDao diagnosed as idiopathic POF,80healthy volunteers over40years old with regularly cycling as control group. DNA was extracted from peripheral blood from POF patients and control subjects, then genomic DNA was used for PCR amplification. The AR gene CAG microsatellite fragment and FMR1gene CGG repeats were identified by DNA sequencing, Results were reported as number of CAG and CGG repeats.105patients were recruited (2009February to2011September) from the gynecological endocrine outpatient clinic of Tianjin Medical University General Hospital and the reproductive medicine clinic of the first hospital of QinHuangDao who diagnosed as idiopathic POF,34patients were infertility among them, and80healthy similar age women with regularly cycling as control group. POF non-infertility patients(7lcases) were divided into two groups randomly:Group A including35patients managed by Estradiol valerate and Cyproterone Acetate tablets (climen), Group B including36patients managed by Premarin0.625mg+MPA6mg cycle treament for three months. POF Infertility patients of POF managed by Estradiol valerate2mg+Progestin capsule200mg cycle treatment for three months. Circulating hormonal parameters, blood lipids profile and fasting blood glucose (FBG) were compared before and after treatment. Infertility patients of POF were induced ovulation with HMG-HCG when FSH level is under15IU/L.62patients were recruited (2009February to2010November) from the reproductive medicine clinic of the first hospital of QinHuangDao diagnosed as idiopathic POF,60healthy similar age women with regularly cycling as control group. POF patients were divided into two groups randomly:Group C include31patients managed by Estradiol valerate and Cyproterone Acetate tablets (climen), Group D also include31patients managed by Premarin0.625mg+MPA6mg cycle treatment for one year. BMD and index of bone metabolism were compared before and after treatment.Results:The AR-CAG repeats ranged from11to34in POF patients and from13to33in control subjects,23and22was the maximum frequency allele in POF and control group. The weighted mean of CAG repeats in POF (22.73±3.24) was significantly higher than in control group (22.03±3.06). The longer CAG repeat length was significant enrichment in POF patients than in control group (P<0.05).The FMR1-CGG repeats ranged from12to42in POF patients and from10to38in control subjects,28was the maximum frequency allele in two groups. The weighted mean of CGG repeats in POF (28.06±4.07) was significantly higher than in control group (26.93±4.37). The intermediate size CGG repeats was significant enrichment in POF patients than in control group (P<0.05).There was no significant difference in blood lipids profile between POF and control group. POF patients were divided into two groups according to amenorrhea time: Group I (<6months) and Group II (>6months) The level of TC and LDL in group Ⅱ were significantly higher than that in group I and control group (P<0.05). The level of HDL in group Ⅱ was significantly lower than that in group I and control group (P<0.05), there was no significant difference in the level of TG and FBG.The estrogen low symptoms were improved after different HRT regimens, the level of FSH and LH were significantly lower and the level of estradiol was significantly higher than before treatment. There were no significant difference of AMH, PRL, T, FBG and blood lipids profile between before and after HRT treatment.20patients of Group Ⅱ managed by climen, and22patients managed by Premarin0.625mg+MPA6mg cycle treatment. The level of TC and LDL after climen treatment were significantly lower than Premarin+MPA treatment (P<0.05), the level of HDL was significantly higher after treatment. There were no significant different FBG and blood lipids profile between before and after Premarin+MPA treatment in Group Ⅱ.There was no significant difference of the Ultrasonic testing index of POF between before and after three cycles HRT. After6cycles, uterine volume was increased significantly after treatment, ovarian volume and antra follicle count (AFC) were not significantly changed (P>0.05)There was no significant difference in the level of FSH, LH and E2at each age of control group. The serum AMH was decreased with age, the level of AMH was significantly lower than the other two groups (P<0.05), AFC was lower than the other two groups (P<0.05)One POF infertility patient was pregnancy at the third cycle of HRT,27patients accepted ovulation induction with the regimens of HMG-HCG after three or six HRT cycles, the follicle could grow to equal or greater than18mm at11cases, and8cases were ovulated and one case was conceived. The follicle of other16cases less than16mm was unpregnancy.6cases were suggested to accept occyte donation IVF-ET as the serum FSH was higher than15IU/L after6cycles HRT.In women with POF, the BMD on lumbar2-4and femoral neck and the level of25-OH-VD3,1,25-OH2-VD3were significantly lower than the control group (P<0.05). The level of osteocalc was significantly higher than the control group (P<0.05). The BMD at lumbar spine and femur of POF patients presented high positive correction with age and body Mass Index (BMI), and negative correlated with length of estrogen deficiency, serum follicle-stimulating hormone (FSH) and serum BGP. There was no significant correlation with age at menarche and serum luteinizing hormone (LH). Serum estradiol (E2),25-OH-VD3and1,25-OH2-VD3presented positive correction with BMD of lumbar spine and no significant correlation with femur. The BMD, level of25-OH-VD3and1,25-OH2-VD3were improved significantly in Group C and D after hormone treatment (P<0.05) Incidence of side-effects was similar in the two groups of patients.Conclusions:1. The CAG repeat length is increased in women with POF as compared with controls, and may be pathogenic for POF, at least in a subset of Chinese han women.2. The number of CGG repeat was significant higher in patients than that in controls but it is in normal range. The relationship of FMR1-CGGn and POF needs deeply study by multicenter and large sample.3. The intermediate sized CGG repeats of FMRl gene is correlated with POF, The mutation of FMR1gene is not find in this study, The mutation is not probably correlated with POF.4. The incidence of abnormal lipid metabolism is increased in POF patient of amenorrhea over six months. Climen has better effect in POF patient’s lipid metabolism.5. AMH is a good index to reflect ovarian function reserves, POF infertility patient can be assessed with serum AMH and AFC and select the feasible ART. 6. POF patient has the possibility of pregnancy during HRT.7. The BMD in POF patient is decreased, the possibility of osteoporosis is increased at future date.8. HRT can improve the bone metabolism of POF patient.