IVF周期血清总睾酮与卵巢反应性参数及颗粒细胞FSHRmRNA表达的相关性研究及睾酮对体外培养人黄素化颗粒细胞FSHRmRNA、IGF-ImRNA、IGF-IImRNA表达的影响
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摘要
研究背景
在颗粒细胞内,雄激素在FSH诱导的芳香化酶作用下转化为雌激素。同时,雄激素又能增加颗粒细胞对FSH的敏感性,促进卵泡发育,如：增加颗粒细胞FSH受体的表达,增加FSH对早期卵泡的募集,促进胰岛素样生长因子-Ⅰ的分泌,IGF-Ⅰ与FSH产生协同作用,促进卵泡生长发育。近年来,国外有些临床研究在IVF周期前或周期过程中使用雄激素处理方案(雄激素启动,androgen priming),包括使用睾酮,脱氢表雄酮,黄体生成素,人绒毛膜促性腺激素和芳香化酶抑制剂等,以提高体循环或卵巢局部的雄激素水平,希望增高的雄激素水平能增加卵巢对外源性促性腺激素的敏感性,增加卵巢反应性,改善IVF结局。然而临床研究结果并不完全一致。因此,我们设计了这项实验,观察IVF过程中血清总睾酮的动态变化、测定颗粒细胞FSHRmRNA的表达及不同浓度睾酮作用下颗粒细胞FSHRmRNA、IGF-ⅠmRNA、IGF-ⅡmRNA的表达,初探IVF周期雄激素处理方案改善卵巢反应性的可行性。
研究目的
1了解IVF周期血清总睾酮与卵巢反应性参数及IVF结局的关系；
2分析IVF周期颗粒细胞FSHRmRNA表达与卵巢反应性及血清总睾酮的关系；
3睾酮对体外培养人黄素化颗粒细胞FSHRmRNA, IGF-ⅠmRNA, IGF-ⅡImRNA基因表达的影响。
研究方法
研究过程分为三部分进行：
第一部分：IVF常规长方案患者进入周期后,监测卵巢刺激过程中血清总睾酮浓度变化,记录患者每次检查时的卵泡数量及大小、血清性激素、取卵数、成熟卵数、优质胚胎数、囊胚数及妊娠情况及使用rFSH剂量、卵巢刺激天数。观察卵巢刺激后血清总睾酮的浓度变化；分别以是否妊娠和血清基础总睾酮分组,分析基础总睾酮与卵巢反应性及IVF结局的关系；探讨血清总睾酮与卵巢反应性参数的相关性。
第二部分：IVF短方案患者进入周期,临床资料的获取方法同第一部分。取卵同时,留取卵泡液,分离颗粒细胞,提取细胞总RNA,反转录cDNA,进行real-time PCR。按照获得的成熟卵数将患者分为卵巢过度反应组、正常反应组和低反应组。比较三组患者的一般情况、卵巢反应性参数及妊娠率、囊胚冷冻率；比较三组血清总睾酮水平及颗粒细胞FSHRmRNA相对表达量,并对血清总睾酮、卵巢反应性参数和FSHRmRNA相对表达量进行相关分析,进一步探讨血清总睾酮、FSHRmRNA表达与卵巢反应性间的关系。
第三部分：取卵时获得人黄素化颗粒细胞,以10-8mol／L、10-7mol／L.10-5mol／L浓度睾酮进行刺激。各组分别在睾酮刺激6小时和24小时时终止培养,提取颗粒细胞总RNA,反转录cDNA,进行real-time PCR。以10-8mol／L作为对照组,比较不同浓度睾酮、刺激不同时间对颗粒细胞FSHRmRNA、IGF-ⅠmRNA、IGF-ⅡmRNA表达的影响。
结果
1第一部分
①卵巢刺激过程中,血清总睾酮浓度随着卵泡的发育逐渐上升；
②分别以血清基础总睾酮(bTT)≤0.2ng/ml和>0.2ng／ml将患者分组,两组间卵巢反应性参数及IVF结局比较均无显著性差异；同样方法再以bTT≤0.3ng／ml和>0.3ng／ml、bTT≤0.4ng/ml和>0.4ng／ml将患者分组,两组间卵巢反应性参数及IVF结局比较仍然均无显著性差异；
③药物刺激卵巢过程中的总睾酮浓度变化与基础总睾酮浓度有平行关系；
④血清基础总睾酮与卵巢反应性参数无线性相关关系；药物刺激卵巢过程中,随着卵泡发育,总睾酮水平与基础窦卵泡数、取卵数、成熟卵泡数有正相关关系,与基础FSH值和rFSH剂量有负相关关系；
⑤妊娠组OPU14日总睾酮值显著高于未妊娠组。
2第二部分
①按照卵巢反应性分组研究发现：卵巢刺激过程中,卵巢对促性腺激素的反应性与颗粒细胞FSHRmRNA的表达量有关,卵巢过度反应组颗粒细胞FSHRmRNA表达量显著高于卵巢低反应组颗粒细胞FSHRmRNA表达量；
②颗粒细胞FSHRmRNA表达量与rFSH用药剂量呈负相关关系,与E2峰、取卵数呈正相关关系；
③血清基础总睾酮与卵巢反应性参数如rFSH剂量、卵巢刺激天数及E2无线性相关关系,与颗粒细胞FSHRmRNA的表达也无线性相关关系；药物刺激卵巢过程中,随着卵泡发育,动态变化的总睾酮水平与rFSH剂量、卵巢刺激天数呈现负相关关系,与E2峰值、取卵数呈现正相关关系,与FSHRmRNA的表达呈现正相关关系。
3第三部分
①不同浓度睾酮刺激颗粒细胞6小时,显著增加颗粒细胞FSHRmRNA的表达,并呈现浓度依赖关系：睾酮浓度越高,基因表达量越高；刺激时间延长至24小时,各组间颗粒细胞FSHRmRNA的表达无统计学差异；
②不同浓度睾酮刺激颗粒细胞6小时,各组间颗粒细胞IGF-ⅠmRNA的表达无统计学差异；刺激时间延长至24小时,颗粒细胞IGF-ⅠmRNA的表达显著降低,并呈现浓度依赖关系：睾酮浓度越高,基因表达量越低；
③不同浓度睾酮刺激颗粒细胞6小时,各组间颗粒细胞IGF-ⅡmRNA的表达无统计学差异；刺激时间延长至24小时,10-7mol／L处理组IGF-ⅡmRNA基因表达量显著高于10-8mol／L处理组和10-5 mol／L处理组。
结论
1基础总睾酮水平在正常范围内的患者,其基础总睾酮水平不能作为预测卵巢反应性及IVF结局的指标；
2 IVF药物刺激卵巢过程中的总睾酮水平与卵巢反应性正相关；颗粒细胞FSHRmRNA表达量与卵巢反应性正相关。
3高浓度睾酮短时间作用能上调FSHR和IGF-Ⅱ基因表达,但长时间作用对基因表达上调作用消失。高浓度睾酮下调IGF-Ⅰ基因表达,随着作用时间的延长,下调作用更显著。
[Background]
Androgens play important roles in follicular growth, development, and atresia, as well as in steroidogenesis. First, androgens act as essential substrates for the production of estrogen. Second, results of some animal experiments showed that Androgens appear to be positive regulators of follicular development by increasing the expression of FSHR and promoting the secretion of the IGF-Ⅰin the granulose cells. IGF-Ⅰshowed some synergistic effects with FSH on follicuogeneisis. Therefore androgens may enhance follicular responsiveness and then promote the development of the follicles. At the same time, animal research also suggested that the androgen might have negative effect on the follicle development. In recent years, it has been reported that some clinical trials used the androgen priming protocol before or during the controlled ovarian hyperstimulation (COH) in IVF patients in order to increase the ovary response to the exogenous gonadotrophins. However, the outcomes of these clinical studies did not accord with each other. Our study was designed to explore the feasibility of androgen supplement before or during the IVF cycle to impove the IVF outcome.
[Objective]
1 To observe the serum total testosterone level during the controlled ovarian stimulation. To assess possible associations between total testosterone level and IVF stimulation parameters or IVF pregnancy outcome.
2 To evaluate the relationship between the serum total testosterone level and the expression of the FSHR in the granulosa cells.
3 To evaluate the effects of testosterone treatment for different doses and different incubating time on FSHR, IGF-Ⅰand IGF-Ⅱexpression in human luteinized granulusa cells.
[Methods]
Our study was devided into three parts: Part one:
A clinical prospective study was involved in this part. The serum total testosterone levels was measured during the IVF cycle. The quantity and the size of the follicles, the levels of sex hormones, and the amount of oocyte retrieved, mature oocyte, and blastomeres were recorded. Also the duration of controlled ovarian stimulation and the total rFSH doses were noted. The patients were devided into two groups according to the IVF outcome or the basal total testosterone level respectively. We described the changes of total testosterone level during the IVF cycle. Linear regression analysis was used to assess the correlation between linear data.
Part two:
Hormone levels and the ovarian response variables were recorded as in part one. In addition, preovulatory granulosa cells were collected from follilcular fluid obtained from patients undergoing transvaginal oocyte retrieval with untrasound guidance. According to the quantity of mature oocyte, the patients were grouped as normal response group, low response group and high response group. The mRNA level of FSHR in the granulosa cells from each patient was quantitatively measured using real-time reverse-transcriptase polymerase chain reaction (RT-PCR). The clinical parameters and the expression of FSHRmRNA among the three groups were compared. And finally, linear regression analysis assessed the correlation between linear data.
Part three:
Preovulatory granulosa cells were collected from follilcular fluid obtained from patients undergoing transvaginal oocyte retrieval with untrasound guidance. After that, granulosa cells were cultured for 2 days in DMEM/F12 medium supplemented with 10%FBS, penicillin/streptomycin, and glutamine. The cells were grouped and incubated in DMEM/F12 medium supplemented with 1%FBS and different concentrations of testosterone (10-8 mol/L,10-7 mol/L,10-5mol/L) for another 6 or 24 hours. The mRNA levels of FSHR, IGF-I and IGF-II were were quantitatively measured using real-time reverse-transcriptase polymerase chain reaction (RT-PCR). We analyzed the effects of testosterone treatment with different doses and different cultured time on FSHR, IGF-I and IGF-II expression in granulusa cells.
[Results]
Part one:
1 During the controlled ovarian stimulation (COH), total testosterone (TT) levels rised in accordance with the follicle development.
2 The change of TT level during the COH presented a parallel phenomenon with the basal TT level.
3 Patients were grouped by basal TT levels at increments of 0.1ng/ml from 0.2ng/ml to 0.4ng/ml. The basal TT levels did not significantly correlate with any of the stimulation parameters tested and the IVF outcome for each increment.
4 TT levels during the COH correlated positively with the antral follicle count, the quantity of oocyte retrieved and mature oocyte. And it correlated negatively with the basal FSH level and the number of gonadrotropin ampoules used.
5 TT level measured 14 days after oocyte retrivel in pregnancy group was significantly higher than that in the non-pregnancy group.
Part two:
1 The expression of FSHR was different among the three different ovarian response groups. FSHRmRNA level s in high response group was significantly higher than that in the low response group.
2 The FSHRmRNA correlated negatively with the total doses of the rFSH. And it correlated positively with the peak E2 level on the day of hCG injection and oocyte number retrieved.
3 The basal TT levels did not significantly correlate with both any of the stimulation parameters tested and the expression of FSHRmRNA. TT levels during the COH correlated negatively with the duration of COH and the number of gonadrotropin ampoules used. And it correlated positively with the peak E2 on the day of hCG injection, the quantity of oocyte retrieved, and the expression of FSHRmRNA.
Part three:
1 After the cells were incubated with testosterone of different concentrations for 6 hours, the expression of FSHRmRNA was significantly increased in a dose-depended way. However, when cultured for 24 hours, no statistical significant difference was observed among the three groups.
2 After the cells were incubated with testosterone of different concentrations for 6 hours, no statistically significant difference were observed on the expression of IGF-ImRNA among the three groups. However, when cultured for 24 hours, the expression of IGF-ImRNA was significantly decreased in a dose-depended way.
3 After the cells were incubated with testosterone of different concentrations for 6 hours, there were no statistically significant differences on the expression of IGF-IImRNA among the three groups. However, when cultured for 24 hours, the expression of IGF-IImRNA in 10-7 mol/L group was higher than that of the 10-8 mol/L group and 10-5mol/L group.
[Conclusions]
1:Basal serum total testosterone level may not predict the IVF outcome and the ovarian response.
2:Total testosterone levels during the controlled ovarian hyperstimulation procedure correlated positively with the ovarian response. The expression of FSHRmRNA in human luteized granulosa cells correlated positively with the ovarian response.
3:Testosterone of high concentration for short time could up-regulate the expression of FSHR and IGF-II. But when treated for long time, the up-regulation of the expression disappeared. Testosterone of high concentration could down-regulate the expression of IGF-I, and the down-regulation became stronger as treated for a longer time.
[Key words]
IVF, testosterone, ovarian response, granulosa cell, FSHRmRNA、IGF-ⅠmRNA、IGF-ⅡmRNA
在颗粒细胞内,雄激素在FSH诱导的芳香化酶作用下转化为雌激素。同时,雄激素又能增加颗粒细胞对FSH的敏感性,促进卵泡发育,如：增加颗粒细胞FSH受体的表达,增加FSH对早期卵泡的募集,促进胰岛素样生长因子-Ⅰ的分泌,IGF-Ⅰ与FSH产生协同作用,促进卵泡生长发育。近年来,国外有些临床研究在IVF周期前或周期过程中使用雄激素处理方案(雄激素启动,androgen priming),包括使用睾酮,脱氢表雄酮,黄体生成素,人绒毛膜促性腺激素和芳香化酶抑制剂等,以提高体循环或卵巢局部的雄激素水平,希望增高的雄激素水平能增加卵巢对外源性促性腺激素的敏感性,增加卵巢反应性,改善IVF结局。然而临床研究结果并不完全一致。因此,我们设计了这项实验,观察IVF过程中血清总睾酮的动态变化、测定颗粒细胞FSHRmRNA的表达及不同浓度睾酮作用下颗粒细胞FSHRmRNA、IGF-ⅠmRNA、IGF-ⅡmRNA的表达,初探IVF周期雄激素处理方案改善卵巢反应性的可行性。
研究目的
1了解IVF周期血清总睾酮与卵巢反应性参数及IVF结局的关系；
2分析IVF周期颗粒细胞FSHRmRNA表达与卵巢反应性及血清总睾酮的关系；
3睾酮对体外培养人黄素化颗粒细胞FSHRmRNA, IGF-ⅠmRNA, IGF-ⅡImRNA基因表达的影响。
研究方法
研究过程分为三部分进行：
第一部分：IVF常规长方案患者进入周期后,监测卵巢刺激过程中血清总睾酮浓度变化,记录患者每次检查时的卵泡数量及大小、血清性激素、取卵数、成熟卵数、优质胚胎数、囊胚数及妊娠情况及使用rFSH剂量、卵巢刺激天数。观察卵巢刺激后血清总睾酮的浓度变化；分别以是否妊娠和血清基础总睾酮分组,分析基础总睾酮与卵巢反应性及IVF结局的关系；探讨血清总睾酮与卵巢反应性参数的相关性。
第二部分：IVF短方案患者进入周期,临床资料的获取方法同第一部分。取卵同时,留取卵泡液,分离颗粒细胞,提取细胞总RNA,反转录cDNA,进行real-time PCR。按照获得的成熟卵数将患者分为卵巢过度反应组、正常反应组和低反应组。比较三组患者的一般情况、卵巢反应性参数及妊娠率、囊胚冷冻率；比较三组血清总睾酮水平及颗粒细胞FSHRmRNA相对表达量,并对血清总睾酮、卵巢反应性参数和FSHRmRNA相对表达量进行相关分析,进一步探讨血清总睾酮、FSHRmRNA表达与卵巢反应性间的关系。
第三部分：取卵时获得人黄素化颗粒细胞,以10-8mol／L、10-7mol／L.10-5mol／L浓度睾酮进行刺激。各组分别在睾酮刺激6小时和24小时时终止培养,提取颗粒细胞总RNA,反转录cDNA,进行real-time PCR。以10-8mol／L作为对照组,比较不同浓度睾酮、刺激不同时间对颗粒细胞FSHRmRNA、IGF-ⅠmRNA、IGF-ⅡmRNA表达的影响。
结果
1第一部分
①卵巢刺激过程中,血清总睾酮浓度随着卵泡的发育逐渐上升；
②分别以血清基础总睾酮(bTT)≤0.2ng/ml和>0.2ng／ml将患者分组,两组间卵巢反应性参数及IVF结局比较均无显著性差异；同样方法再以bTT≤0.3ng／ml和>0.3ng／ml、bTT≤0.4ng/ml和>0.4ng／ml将患者分组,两组间卵巢反应性参数及IVF结局比较仍然均无显著性差异；
③药物刺激卵巢过程中的总睾酮浓度变化与基础总睾酮浓度有平行关系；
④血清基础总睾酮与卵巢反应性参数无线性相关关系；药物刺激卵巢过程中,随着卵泡发育,总睾酮水平与基础窦卵泡数、取卵数、成熟卵泡数有正相关关系,与基础FSH值和rFSH剂量有负相关关系；
⑤妊娠组OPU14日总睾酮值显著高于未妊娠组。
2第二部分
①按照卵巢反应性分组研究发现：卵巢刺激过程中,卵巢对促性腺激素的反应性与颗粒细胞FSHRmRNA的表达量有关,卵巢过度反应组颗粒细胞FSHRmRNA表达量显著高于卵巢低反应组颗粒细胞FSHRmRNA表达量；
②颗粒细胞FSHRmRNA表达量与rFSH用药剂量呈负相关关系,与E2峰、取卵数呈正相关关系；
③血清基础总睾酮与卵巢反应性参数如rFSH剂量、卵巢刺激天数及E2无线性相关关系,与颗粒细胞FSHRmRNA的表达也无线性相关关系；药物刺激卵巢过程中,随着卵泡发育,动态变化的总睾酮水平与rFSH剂量、卵巢刺激天数呈现负相关关系,与E2峰值、取卵数呈现正相关关系,与FSHRmRNA的表达呈现正相关关系。
3第三部分
①不同浓度睾酮刺激颗粒细胞6小时,显著增加颗粒细胞FSHRmRNA的表达,并呈现浓度依赖关系：睾酮浓度越高,基因表达量越高；刺激时间延长至24小时,各组间颗粒细胞FSHRmRNA的表达无统计学差异；
②不同浓度睾酮刺激颗粒细胞6小时,各组间颗粒细胞IGF-ⅠmRNA的表达无统计学差异；刺激时间延长至24小时,颗粒细胞IGF-ⅠmRNA的表达显著降低,并呈现浓度依赖关系：睾酮浓度越高,基因表达量越低；
③不同浓度睾酮刺激颗粒细胞6小时,各组间颗粒细胞IGF-ⅡmRNA的表达无统计学差异；刺激时间延长至24小时,10-7mol／L处理组IGF-ⅡmRNA基因表达量显著高于10-8mol／L处理组和10-5 mol／L处理组。
结论
1基础总睾酮水平在正常范围内的患者,其基础总睾酮水平不能作为预测卵巢反应性及IVF结局的指标；
2 IVF药物刺激卵巢过程中的总睾酮水平与卵巢反应性正相关；颗粒细胞FSHRmRNA表达量与卵巢反应性正相关。
3高浓度睾酮短时间作用能上调FSHR和IGF-Ⅱ基因表达,但长时间作用对基因表达上调作用消失。高浓度睾酮下调IGF-Ⅰ基因表达,随着作用时间的延长,下调作用更显著。
[Background]
Androgens play important roles in follicular growth, development, and atresia, as well as in steroidogenesis. First, androgens act as essential substrates for the production of estrogen. Second, results of some animal experiments showed that Androgens appear to be positive regulators of follicular development by increasing the expression of FSHR and promoting the secretion of the IGF-Ⅰin the granulose cells. IGF-Ⅰshowed some synergistic effects with FSH on follicuogeneisis. Therefore androgens may enhance follicular responsiveness and then promote the development of the follicles. At the same time, animal research also suggested that the androgen might have negative effect on the follicle development. In recent years, it has been reported that some clinical trials used the androgen priming protocol before or during the controlled ovarian hyperstimulation (COH) in IVF patients in order to increase the ovary response to the exogenous gonadotrophins. However, the outcomes of these clinical studies did not accord with each other. Our study was designed to explore the feasibility of androgen supplement before or during the IVF cycle to impove the IVF outcome.
[Objective]
1 To observe the serum total testosterone level during the controlled ovarian stimulation. To assess possible associations between total testosterone level and IVF stimulation parameters or IVF pregnancy outcome.
2 To evaluate the relationship between the serum total testosterone level and the expression of the FSHR in the granulosa cells.
3 To evaluate the effects of testosterone treatment for different doses and different incubating time on FSHR, IGF-Ⅰand IGF-Ⅱexpression in human luteinized granulusa cells.
[Methods]
Our study was devided into three parts: Part one:
A clinical prospective study was involved in this part. The serum total testosterone levels was measured during the IVF cycle. The quantity and the size of the follicles, the levels of sex hormones, and the amount of oocyte retrieved, mature oocyte, and blastomeres were recorded. Also the duration of controlled ovarian stimulation and the total rFSH doses were noted. The patients were devided into two groups according to the IVF outcome or the basal total testosterone level respectively. We described the changes of total testosterone level during the IVF cycle. Linear regression analysis was used to assess the correlation between linear data.
Part two:
Hormone levels and the ovarian response variables were recorded as in part one. In addition, preovulatory granulosa cells were collected from follilcular fluid obtained from patients undergoing transvaginal oocyte retrieval with untrasound guidance. According to the quantity of mature oocyte, the patients were grouped as normal response group, low response group and high response group. The mRNA level of FSHR in the granulosa cells from each patient was quantitatively measured using real-time reverse-transcriptase polymerase chain reaction (RT-PCR). The clinical parameters and the expression of FSHRmRNA among the three groups were compared. And finally, linear regression analysis assessed the correlation between linear data.
Part three:
Preovulatory granulosa cells were collected from follilcular fluid obtained from patients undergoing transvaginal oocyte retrieval with untrasound guidance. After that, granulosa cells were cultured for 2 days in DMEM/F12 medium supplemented with 10%FBS, penicillin/streptomycin, and glutamine. The cells were grouped and incubated in DMEM/F12 medium supplemented with 1%FBS and different concentrations of testosterone (10-8 mol/L,10-7 mol/L,10-5mol/L) for another 6 or 24 hours. The mRNA levels of FSHR, IGF-I and IGF-II were were quantitatively measured using real-time reverse-transcriptase polymerase chain reaction (RT-PCR). We analyzed the effects of testosterone treatment with different doses and different cultured time on FSHR, IGF-I and IGF-II expression in granulusa cells.
[Results]
Part one:
1 During the controlled ovarian stimulation (COH), total testosterone (TT) levels rised in accordance with the follicle development.
2 The change of TT level during the COH presented a parallel phenomenon with the basal TT level.
3 Patients were grouped by basal TT levels at increments of 0.1ng/ml from 0.2ng/ml to 0.4ng/ml. The basal TT levels did not significantly correlate with any of the stimulation parameters tested and the IVF outcome for each increment.
4 TT levels during the COH correlated positively with the antral follicle count, the quantity of oocyte retrieved and mature oocyte. And it correlated negatively with the basal FSH level and the number of gonadrotropin ampoules used.
5 TT level measured 14 days after oocyte retrivel in pregnancy group was significantly higher than that in the non-pregnancy group.
Part two:
1 The expression of FSHR was different among the three different ovarian response groups. FSHRmRNA level s in high response group was significantly higher than that in the low response group.
2 The FSHRmRNA correlated negatively with the total doses of the rFSH. And it correlated positively with the peak E2 level on the day of hCG injection and oocyte number retrieved.
3 The basal TT levels did not significantly correlate with both any of the stimulation parameters tested and the expression of FSHRmRNA. TT levels during the COH correlated negatively with the duration of COH and the number of gonadrotropin ampoules used. And it correlated positively with the peak E2 on the day of hCG injection, the quantity of oocyte retrieved, and the expression of FSHRmRNA.
Part three:
1 After the cells were incubated with testosterone of different concentrations for 6 hours, the expression of FSHRmRNA was significantly increased in a dose-depended way. However, when cultured for 24 hours, no statistical significant difference was observed among the three groups.
2 After the cells were incubated with testosterone of different concentrations for 6 hours, no statistically significant difference were observed on the expression of IGF-ImRNA among the three groups. However, when cultured for 24 hours, the expression of IGF-ImRNA was significantly decreased in a dose-depended way.
3 After the cells were incubated with testosterone of different concentrations for 6 hours, there were no statistically significant differences on the expression of IGF-IImRNA among the three groups. However, when cultured for 24 hours, the expression of IGF-IImRNA in 10-7 mol/L group was higher than that of the 10-8 mol/L group and 10-5mol/L group.
[Conclusions]
1:Basal serum total testosterone level may not predict the IVF outcome and the ovarian response.
2:Total testosterone levels during the controlled ovarian hyperstimulation procedure correlated positively with the ovarian response. The expression of FSHRmRNA in human luteized granulosa cells correlated positively with the ovarian response.
3:Testosterone of high concentration for short time could up-regulate the expression of FSHR and IGF-II. But when treated for long time, the up-regulation of the expression disappeared. Testosterone of high concentration could down-regulate the expression of IGF-I, and the down-regulation became stronger as treated for a longer time.
[Key words]
IVF, testosterone, ovarian response, granulosa cell, FSHRmRNA、IGF-ⅠmRNA、IGF-ⅡmRNA
引文
[1]Martin KA,Hornstein MD,Taylor AE,et al.Exogenous gonadotropin stimulation is associate with increases in serum androgen levels in in-vitro fertilization-embryo transfer cycles.Fertil Steril,1997;68(6):1011-1016
[2]Cedars MI,Surey E,Hamilton F,et al.Leuprolide acetate lowers circulating bioactive luteinizing hormone and testosterone concentrations during ovarian stimulation for oocyte retrieval.Fertil Steril.1990;53(4):627-631
[3]Fanchin R,de Ziegler D,Castracane VD,et al.Physiopathology of premature progesterone elevation.Fertil Steril 1995;64(4):796-801
[4]Orvieto R,Yulzari-Roll V,La Marca A,et al.Serum androgen levels in patients undergoing controlled ovarian hyperstimulation for in vitro fertilization cycles.Assisted Repred Technology.2005,21(4):218-222
[5]Andersen CY,Ziebe S.Serum levels of free androstenedione,testosterone and oestradiol are lower in the follicular phase of conceptional than of non-conceptional cycles after ovarian stimulation with a gonadotrophin-rel easing hormone agonist protocol.Hum Reprod.1992,7(10):1365-1370
[6]Check JH,Nazari A,Dietterich C.Comparison of androgen levels in conception vs.non-conception cycles following controlled ovarian stimulation using the luteal phase gonadotropin-releasing hormone agonist protocol. Gynecol Endocrinol.1995;9(3):209-214
[7]Frattarelli JL,Gerber MD.Basal and cycle androgen levels correlate with in vitro fertilization stimulation parameters but do not predict pregnancy outcome.Fertil Steril,2006,86(1):51-57
[8]Frattarelli JL,Peterson EH.Effect of androgen levels on in vitro f ertilization cycles.Fertility Sterility 2004,81 (6):1713-1714
[9]Tanbo T,Dale PO,Abyholm T.Recombinant follicle-stimulating hormone stimulates ovarian androgen synthesis in down-regulated ovulatory women.Gynecol Edocrinol.2001;15(6):407-412
[10]Tarlatzis BC,Zepiridis L,Grimbizis G,et al.Clinical management of low ovarian response to stimulation for ⅣF:a systematic review.Hum Reprod Update.2003,9(1):61-76
[1]Salas C,Julio-Pieper M,Valladares M, et al.Nerve growth factor-dependent activation of takA receptors in the human ovary results in synthesis of follicle-stimulating hormone receptors and estrogen secretion.J Clinic Endocrino,2006,91 (6):2396-2403.
[2]Tarlatzis BC,Zepiridis L,Grimbizis G,et al.Clinical management of low ovarian response to stimulation for IVF:systematic review[J].Hum Reprod Update,2003,9(1):61-76
[3]O'Shaughenssy PJ,Dudley K,Rajapaksha WR.Expression of follicle stimulating hormone-receptor mRNA during gonadal development.Mol Cell Endocrinol,2004; 125(1-2_:169-175
[4]Bogovich K.Obligatory roles for follicle-stimulating hormone(FSH),estradiol and androgens in the induction of small pilyfollicular ovarian cysts in hypophysectomized immature rats.Endocrine,2007,31 (2):179-192
[5]Scott RT,Opsahl MS,Leonardi MR,et al.Life table analysis of pregnancy rates in a general infertility population relative to ovarian reserve and patient age.Hum Reprod 1995;10(7):1706-1710
[6]Huhtaniemi IT.LH and FSH receptor mutations and their effects on puberty.Horm Res,2002,57(suppl 2):35-38
[7]Abel MH,Wootton AN,Wilkins V,et al.The effect of a null mutation in the follicle-stimulating hormone receptor gene on mouse reprodution.Endocrinology,2003,141 (5):1795-1803
[8]Pellicer A,Ballester MJ,Serrano MD,et al.Aetiological factors involved in the low response to gonadotrophins in infertile women with normal basal. serum follicle stimulating hormone levels.Hum Reprod 1994;9(5):806-811
[9]Abdennebi L,Monget P,Pisselet C,et al.Comparative expression of luteinizing hormone and follicle-stimulating hormone receptors in ovarian follicles from high and low prolific sheep breeds.Biol Reprod,1999;60(4):845-854
[10]Loutradis D,Patsoula E,Minas V,et al.FSH receptor gene polymorphisms have a role for different ovarian response to stimulation in patients entering IVF/ICSI-ET programs.J Assist Reprod Genet,2006,23(4):177-184
[11]楼航英,黄荷凤。超排卵时卵巢对促性腺激素的反应与颗粒细胞促卵泡激素受体的表达。浙江大学学报,2002,31(3)：151-154
[12]蔡婕,高惠娟,黄荷凤。超排卵周期卵巢的反应性与卵泡颗粒细胞中卵泡刺激素受体表达的关系。中华妇产科杂志。2006,41(6)：372-374
[13]Cai J,Lou HY,Dong MY,et al.Poor ovarian response to gonadotropin stimulation associated with low expression of follicle-stimulating hormone receptor in gramulosa cells.Fertil Steril,2007;87(6):1350-1356
[14]Wang H,Andoh K,Hagiwara H,et al.Effect of adrenal and ovarian androgens on type 4 follicles unresponsive to FSH in immature mice[J].Endocrinoogy,2001,142 (11):4930-4936
[15]Vendola KA,Zhou J,Adesanya OO,et al.Androgens stimulate early stages of follicular growth in the primate ovary[J].J Clin Invest,1998,101(12):2622-2629
[16]Cardenas H,Herrick JR,Pope WF.Increased ovulation rate in gilts treated with dihydrotestosterone.Reproduction.2002,123(4):527-533
[17]Casson P R, Lindsay MS, Pisarska MD,et al.Dehydroepiandrosterone supplementation augments ovarian stimulation in poor responders:a case series.Hum Reprod,2000,15(10):2129-2132
[18]Fabregues F,Penarrubia J,Creus M,et al.Transdermal testosterone may improve ovarian response to gonadotrophins in low-responder IVF patient:a randomized,clinical trial.Hum Reprod,2009,24(2):349-359
[19]Lossl K,Andersen AN,Loft A,et al.Androgen priming using aromatase inhibitor and hCG during early-follicular-phase GnRH antagonist down-regulation in modified antagonist protocols.Human Reprod,2006,21(10):2593-2600
[20]Massin N.Cedrin-Durnerin I,Coussieu C,et al.Effects of transdermal testosterone application on the ovarian response to FSH in poor responders undergoing assisted reproduction technique-a prospective,randomized double-blind study.Hum Reprod,2006,21(5):1204-1211
[21]Weil S,Vendola K,Zhou J,et al.Androgen and follicle-stimulating hormone interactions in primate ovarian follicle development. J Clin Endocrinol Metab.1999,84(8):2951-2956
[22]Catteau-Jonard S,Jamin SP,Leclerc A,et al.Anti-Mullerian Hormone,its receptor,FSH receptor,and androgen receptor genes are overexpressed by granulosa cells from stimulated follicles in women with polycystic ovary syndrome.J Clin Endocrinol metab.2008,93(11):4456-4461
[23]Simon C,Garcia Velasco JJ,Valbuena D,et al.Increasing uterine receptivety decresasing estradiol levels during the preimplantation period in high responders with the use of a follicle-stimulating hormone step-down regimen.Fertil Steril,1998,70(2):234-239
[24]Simon C,Domingguez F,Valbuena D,et al.The role of estrogen in uterine receptivity and blastecyst implantation.Trends Endtrinol,Metab,2003,14(5):197-199
[25]Forman R,Fries N,Testart J,et al.Evidence for an adverse effect of elevated serum estradiol concentrations on embryo implantation.Fertil Steril,1988,49(1):118-122 [26]Sharara F,McClamrock HD.High estradiol levels and high oocyte yield are not detrimental to in vitro fertilization outcome.Fertil Steril,1999,72(3):401-405
[1]Lossl K, Andersen AN, Loft A,et al.Androgen priming using aromatiase inhibitor and hCG during early-follicular-phase GnRH antagonist down-regulation in modified antagonist protocols.2006,21(10):2593-2600
[2]Greisen S, Ledet T, Ovesen P.Effects of androstenedione,insulin and luteinizing hormone on steroidogenesis in human granulose luteal cells.Human Reprod,2001;16(10):2061-65
[3]C Ganeff,G Chatel,Munaut,et al. The IGF system in in-vitro human dicidualization,2009,15(1):27-38
[4]Wang HS,Chard T.The role of insulin-like growth factor-I and insulinOlike growth factor-binding protein-1 in the control of human fetal growth. J Endocrinol,1992,132(1):11-19
[5]Wang HS,Chard T.IGFs and IGF-binding proteins in the regulation of human ovarian and endometrial function J Endocrinol,1999,161(1):1-13
[6]Rajaram S,Baylink DJ,Mohan S.Insulin-like growth factor-binding proteins in serum and other biological fluids:regulation and functions.Endocr Rev,1997,18(6):801-831
[7]Jones JI,Clemmons DR.Insulin-like growth factors and their binding proteins:biological actions.Endocr Rev.1995,16(1):3-34
[8]Giudice LC.Insulin-like growth factor family in Graafian follicle development and function.J Soc Gynecol Investig 2001,8(1 Suppl Proceedings):S26-9
[9]Vendola K,Zhou J,Wang J,et al.Androgens rpomote oovyte insulin-like growth factor I expression and initiation of follicle development in the primate ovary. Biol Reprod,1999,61 (2):353-357
[10]Thomas FH,Campbell BK.Armstrong DG,et al.Effects of IGF-I bioavailability on bovine preantral follicular devlopment in vitro.Reproduction,2007,133(6):1121-1128
[11]Demeestere I,Gervy C,Centner J,et al.Effect of insulin-like growth factor-I during preantral follicular culture on steroidogenesis,in vitro oocyte maturation,and embryo development in mice.Biol Reprod,2004,70(6):1664-1669
[12]Monget P,Fabre S,Mulsant P,et al.Regulation of ovarian folliculogenesisby IGF and BMP system in domestic animals.Domest Anim Endocrinol,2002,23(1-2):139-154
[13]Zhou J,Kumar TR,Matzuk MM,et al.Insulin-like growth factor I regulates gonadotropin responsiveness in the murine ovary.Mol Endocrinol, 1997,11 (13):1924-1933
[14]Zhou H,Zhang Y.Effect of growth factors on in vitro developement of caprine preantral follicle oocytes.Anim Reprod Sci,2005,90(3-4):265-272
[15]Monniaux D,Pisselet C.Control of proliferation and differentiation of ovine granulosa cells by insulin-like growth factor-I and follicle-stimulating hormone in vitro.Biol Reprod,1992,46(1):109-119
[16]Louhio H,Hovatta O,Sjoberg J,et al.The effects of insulin,and insulin-like growth factors I and II on human ovarian follicles in long-term culture.Mol Hum Reprod,2000,6(8):694-698.
[17]Hemamalini NC,Rao BS,Tamilmani G,et al.Influence of transforming growth factor-α,insulin-like growth factor-Ⅱ,epidermal growth factor or follicle stimulating hormone on in vitro development of preantral follicles in sheep.Smal Rum Res 2003;50:11-22
[18]Rajarajan K,Rao BS,Vagdevi R,et al.Effect of arious growth factors on the in vitro development of goat preantral follicles.Small Rum Res 2006;63:204-212
[19]Willis DS,Mason HD,Watson H,et al.Developmentally regulated responses of human granulosa cells to insulin-like growth factors (IGFS):IGF-I and IGF-II action mediated via the type-I IGF receptor.J Clin Endocrinol Metab,199883(4):1256-1259
[20]el-Roeiy A,Chen X.Roberts VJ,et al.Expression of insulin-like growth factor-I(IGF-I) and IGF-II and the IGF-I,IGF-Ⅱ and insulin receptor genes and localization of the gene products in the human ovary.J Clin Endocrinol Metab,1993,77(5):1411-1418
[21]Voutilainen R,Franks S,Mason HD,et al.Expression of insulin-like growth factor(IGF),IGF-binding protein and IGF receptor messenger ribonucleic acids in normal and polycystic ovaries.J Clin Endocrinol Metab.1996,81(3):1003-1008
[22]Giudice LC.The insulin-like growth factor system in normal and abnormal human ovarian follicle development.Am J Med.1995,98(1):48S-54S
[23]Jian Zhou,Carolyn B.Anatomy of th human ovarian insulin-like growth factor system.Bioloby of reproduction.1993,48,467-482.
[24]Oliver JE,Aitman JR,Powell JF,et al.IGF-I gene expression in the rat ovary is confined to the granulosa cells of developing follicles.Endocronology,1989; 124(6):2671-2679
[25]Zhou J,Chin E,Bondy C.Cellular patern of IGF-I and IGF-I receptor gene expression in the developing and mature follicle.Endocrinology, 1991:129(6):3281-3288
[26]Chang SY,Tsai MY,Huang FJ,et al.Expression of insulin-like growth factor(IGF),IGF receptor,and IGF-Binding protein messenger ribonucleic acids in luteinized granulosa cells from different size follicles after controlled ovarian hyperstimulatin.J Assist Reprod Genet,2002,19(3):1221-126
[27]Jesionowska H,Hemmings R,Guyda HJ,et al.Determination of insulin and insulin-like growth factors in the ovarian circulation.Fertil Steril,1990,53(1):88-91
[28]Chang SY,Hsieh KC,Wang HS,et al.Follicular fluid levels of insulin-like growth factor I,insulin-like growth factor binding protein I,and ovarian steroids collected during ovum pick-up.Fertil Steril,1994,62(1):1162-1167
[29]Pellegrini S,Fuzzi B.Pratesi S,et al.In-vivo stydies on ovarian insulin-like growth factor I concentrations in human preovulatory follicles and human ovarian circulation.Hum Reprod,1995,10(6):1341-1345
[30]Thierry van Dessel HJ,Chandrasekher Y,Yap OW,et al.Serum and follicular fluid levels of insulin-like growth factor I(IGF-I),IGF-II,and IGF-binding protein-1 and-3 during the normal menstrual cycle.J Clin Endocrinol Metab,1996,81(3):1224-1231
[31]Kubota T,Kamada S,Ohara M,et al.Insulin-like growth factor Ⅱ in follicular fluid of the patients with in vitro fertilization and embryo transfer.Fertil Steril,1993,59(4):844-849
[32]Hickey TE,Marrocco DL,Amato F,et al.Androgens augment the mitogenic effects of oocyte-secreted factors and growth differentiation factor 9 on porcine granulosa cells.Biol Reprod,2005,73(4):825-832
[33]Hickey TE,Marrocco DL,Gilchrist RB,et al.Interactions between androgen and growth factors in granulosa cell subtypes of porcine antral follicles.Biol Reprod,2004:71(1):45-52
[34]el-Roeiy A,Chen X,Roberts VJ,et al.Expression of the genes encoding the insulin-like growth factors(IGF-I and II),the IGF and insulin receptors,and IGF-binding proteins 1-6 and the localization of their gene products in normal and polycystic ovary syndrome ovaries.J Clin Endocrinol Metab,1994,78(6):1488-1496
[35]Ramasharma K,Cabrera CM,Li CH.Identification of insulin-like growth factor-II in human seminal and follicular fluids.Biochem Biophys Res Commun,1986,140(2):536-542
[36]Voutilainen R,Miller WL.Coordinate tropic hormone regulation of mRNAs for insulin-like growth factor Ⅱ and the cholesterol side-chain-cleavage enzyme,P450ssc[corrected],in human steroidogenic tissues.Proc Natl Acad Sci USA,1987,84(6):1590-1594.
[37]Barreca A,Del Monte P,Ponzani P,et al.Intrafollicular insulin-like trowth factor-II levels in normally ovulating women and in patients with polycystic ovary syndrome.Fertil Steril,1996,65(4):739-745.
[1]Tarlatzis BC,Zepiridis L,Grimbizis G,et al.Clinical management of low ovarian response to stimulation for IVF:systematic review[J].Hum Reprod Update,2003,9(1):61-76
[2]Hugues JN,Cedrin Durnerin IC.Revisiting gonadotrophin-releasing hormone agonist protocols and management of poor ovarian responses to gonadotrophins[J].Hum Reprod Update,1998,4(1),83-101
[3]Hofmann GE,Toner JP.Muasher SJ,et al.High-dose follicle stimulationg hormone (FSH)ovarian stimulation in low-responder patients for in vitro fertilization[J].J In Vitro Fert Embryo Transf,1989,6(5),285-289
[4]Check JH,Nowroozi K,Chase JS,et al.Ovulation induction and pregnancies in 100 consecutive women with hypergonadotropic amenorrhea[J].Ferti Steril.1990,53(5):811-6
[5]Homburg R,Eshel A,Abdalla HI,et al.Growth hormone facilitates ovulation induction by gonadotrophins.Clin Endocrionl(Oxf).1988,29(1),113-117
[6]Keay SD,Lenton EA,Cooke ID,et al.Low-dose dexamethasone augments the ovarian response to exogenous gonadotrophins leading to a reduction in cycle cancellation rate in a standard IVF programme[J].Hum Reprod,2001,16(9):1861-1865
[7]Orentreich N.Brind JL,Rizer RL,et al.Age changes and sex differences in serum dehydroepiandrosterone sulfate concentrations throughout adulthood.J Clin Endocrinol Metab.1984,59(3):551-555
[8]Frattarelli JL,Peterson EH.Effect of androgen levels on in vitro fertilization cycles[J]. Fertil Steril.2004,81(6):1713-1714
[9]Orvieto R,Yulzari-Roll V,La Marca A,et al.Serum androgen levels in patients undergoing conrolled ovarian hyperstimulation for in vitro fertilization cycles[J].Gynecol endocrinol.2005,21(4):218-222
[10]Wang H,Andoh K,Hagiwara H,et al.Effect of adrenal and ovarian androgens on type 4 follicles unresponsive to FSH in immature mice[J].Endocrinoogy 2001,142 (11):4930-4936
[11]Walters KA,Allan CM,Handelsman DJ.Androgen actions and the ovary[J].Biol Reprod,2008;78(3):380-389
[12]Vendola KA,Zhou J,Adesanya OO,et al.Androgens stimulate early stages of follicular growth in the primate ovary[J].J Clin Invest,1998,101(12):2622-2629
[13]Ware VC.The role of androgens in follicular development in the ovary.Ⅰ.A quantitative analysis of oocyte ovulation[J] J Exp Zool 1982,222(2):155-167
[14]Cardenas H,Herrick JR,Pope WF.Increased ovulation rate in gilts treated with dihydrotestosterone.Reproduction.2002,123(4):527-533
[15]Beloosesky R,Gold T,Almog B,et al.Induction of polycystic ovary by testosterone in immature female rats modulation of apoptosis and attenuation of glucose/insulin ratio.Int J Mol Med,2004,14(2):207-215
[16]Hillier SG,Whitelaw PF,Smyth CD.Follicular ostrogen synthesis:the'the two-cell,two-gonadotrophin'model revisited[J].Mol Cell Endocrinol 1994,100(1-2):51-54
[17]Quigley CA,De Bellis A,Marschke KB,et al.Androgen receptor defects:historical,clinical,and molecular perspectives.Endocr Rev,1995;16(3):271-321
[18]Hirai M,Hirata S,Osada T,et al.Androgen receptor mRNA in the rat ovary and uterus J Steroid Biochem Mol Biol.1994;49(1):1-7
[19]Tetsuka M,Hillier SGAndrogen receptor gene expression in rat granulosa cells:the role of follicle-stimulating hormone and steroid hormones[J].Endocrinology,1996,137(10):4392-4397
[20]Szoltys M,Slomczynska M.Changes in distribution of androgen receptor during maturation of rat ovarian follicles.Exp Clin Endocrinol Diabetes,2000,108(3):228-234
[21]Juengel JL,Heath DA,Quirke LD,et al.Oestrogen receptor alpha and beta,androgen receptor and progesterone receptor mRNA and protein localisation within the developing ovary and in small growing follicles of sheep[J].Reproduction,2006;131(1):81-92
[22]Hampton JH,Manikkam M,Lubahn DB,et al.Androgen receptor mRNA expression in the bovine ovary.Domest Anim Endocrinol 2004;27(1):81-88
[23]Slomczynska M,Duda M,Sl zak K.The expression of androgen receptor,cytochrome P450 aromatase and FSH receptor mRNA in the porcine ovary.Folia Histochem Cytobiol.2001;39(1):9-13.
[24]Cardenas H,Pope WF.Androgen receptor and follicle-stimulating hormone receptor in the pig ovary during the follicular phase of the estrous cycle.Mol Reprod Dev,2002;62(1):92-98
[25]Hillier SQTetsuka M,Fraser HM.Location and developmental regulation of androgen receptor in primate ovary.Hum Reprod,1997,12(1):107-111
[26]Chaffin CL,Stouffer RL,Duffy DM.Gonadotropin and steroid regulation of steroid receptor and aryl hydrocarbon receptor messenger ribonucleic acid in macaque granulosa cells during the periovulatory interval.Endocrinology,1999;140(10):4753-4760
[27]Duffy DM,Abdelgadir SE,Stott KR,et al.Androgen receptor mRNA expression in the rhesus monkey ovary.Endocrine.1999,11(1):23-30
[28]Horie k,Takakura K,Fujiwara H,et al.Immunohistochemical localization of androgen receptor in the human ovary throughout the menstrual cycle in relation to oestrogen and progesterone receptor expression.Hum Reprod 1992,7(2):184-190
[29]Weil SJ,Vendola K,Zhou J,et al.Androgen receptor gene expression in the primate ovary:cellular localization,regulation,and functional correlations.J Clin Endocrinol Metab.1998,83(7):2479-2485
[30]Catteau-Jonard S,Jamin SP,Leclerc A,et al.Anti-Mullerian Hormone,its receptor,FSH receptor,and androgen receptor genes are overexpressed by granulosa cells from stimulated follicles in women with polycystic ovary syndrome.J Clin Endocrinol metab.2008,93(11):4456-4461
[31]Weil S,Vendola K,Zhou J,et al.Androgen and follicle-stimulating hormone interactions in primate ovarian follicle development. J Clin Endocrinol Metab.1999,84(8):2951-56
[32]Yoshimura Y,Ando M,Nagamatsu S,et al.Effects of insulin-like growth factor-I on follicle growth,oocyte maturation and ovarian steroidogenesis and plasminogen activator anctivity in the rabbit,Biol Reprod,1996,55(1):152-160
[33]Zhou J,Kumar TR,Matzuk MM,et al.Insulin-like growth factor I regulates gonadotropin responsiveness in the murine ovary.Mol Endocrinol.1997;11(13):1924-1933
[34]Hickey TE,Marrocco DL,Amato F,et al.Androgens augment the mitogenic effects of oocyte-secreted factors and growth differentiation factor 9 on porcine granulosa cells.Biol Reprod,2005,73(4):825-832
[35]Hickey TE,Marrocco DL,Gilchrist RB,et al.Interactions between androgen and growth factors in granulosa cell subtypes of porcine antral follicles.Biol Reprod,2004:71(1):45-52
[36]Vendola K,Zhou J,Wang J,et al.Androgens promote oocyte insulin-like growth factor I expression and initiation of follicle development in the primate ovary.Biol Reprod,1999,61(2):353-357
[37]Hamel M,Vanselow J,Nicola ES,et al.Androstenedione increases cytochrome P450 aromatase messenger ribonucleic acid transcripts in nonluteinizing bovine granulosa cells[J].Mol Reprod Dev,2005,70(2):175-183
[38]ElBeltagy K,Honda K,Ozaki K,et al.In vitro effect of dehydroepiandrosterone sulfate on steroid receptors,aromatase,cyclooxygenase-2 expression,and steroid hormone production in preovulatory human granulose cells.Fertil Steril,2007,88(4 suppl):1135-1142
[39]Billig H,Furuta I,Hsueh AJ.Estrogens inhibit and androgens enhance ovarian Granulosa cell apoptosis.Endocrinology 1993;133 (5):2204-2212
[40]Conway BA,Mahesh VB,Mills TM.Effect of dihydrotestoterone on the growth and function of ovarian follicles in intact immature female rats primed with PMSG.J Reprod Fertil,1990;90(1):267-277
[41]Farookhi R.Effects of aromatizable and nonaromatizable androgen treatments on luteinizing hormone receptors and ovulation induction in immature rats.Boil Reprod,1985;33(2):363-369
[42]Azzolin GC,Saiduddin S.Effect of androgens on the ovarian morphology of the hypophysectomized rat.Proc Soc Exp Biol Med,1983,172(1):70-73
[43]张娟,朱桂金,王昕荣,等。多囊卵巢综合征模型鼠颗粒细胞凋亡及TRAIL蛋白的表达。中国组织化学与细胞化学杂志。2007,16(3)：352-356
[44]Zeleznik AJ,Hillier SQRoss GT.Follicle stimulating hormone-induced follicular development:an examination of the role of androgens.Biol Reprod,1979,21(3);673-681
[45]Anderiesz C,Trounson AO.The effect of testosterone on the maturation and developmental capacity of murine oocytes in vitro.Hum Reprod,1995,10(9):2377-2381
[46]Harlow CR,Shaw HJ,Hillier SG,et al.Factors influencing follicle-stimulating hormone-responsive steroidogenesis in marmoset gramulosa cell:effects of androgens and the stage of follicular maturity.Endocrinology,1988;122(6):2780-2787
[47]Zeleznik AJ,Little-lhrig L,Ramasawamy S.Administration of dihydrotestosterone to rhesus monkeys inhibits gonadotropin-stimulated ovarian steroidogenisis[J].J Clin endocrinol metab,2004,89(2);860-866
[48]孙春玲,乔杰,陈英玉。体外受精-胚胎移植中多囊卵巢综合征颗粒细胞凋亡与妊娠结局关系的探讨。中国实用妇科与产科杂志。2007,23(9)：695-697
[49]Weenen C,Laven JS,Von Bergh AR,et al.AntiMullerian homone expression pattern in the human ovary.Potential implications for initial and cyclic follicle recruitment [J],Mol Hum Reprod,2004,10(2):77-83
[50]Durlinger AL, Gruijters MJ, Kramer P, et al. Anti-Mullerian hormone attenuates the effects of FSH on follicle development in the mouse ovary[J]. Endocrinology, 2001,142(11):4891-4899.
[51]Pigny P,Merlen E,Robert Y,et al.Elevated serum level of anti-Mullerian humone in patients with polycystic ovary syndrome:relationship to the ovarian follicle excess and to the follicular arrest[J].J Clin Endocrinol Metab.2003,88(12):5957-5962
[52]Piltonen T,Morin-Papunen L,Koivunen R,et al.Serun anti-Mullerian hormone levels remain high until late reproductive age and decrease during metformin therapu in women with polycystic ovary syndrome[J].Hum Reprod,2005,20(7):1820-1826
[53]Eldar-Geva T, Margalioth EJ, Gai M, et al. Serum anti-Mullerian hormone levels during controlled ovarian hyperstimulation in women in polycystic ovaries with and without hyperandrogenism[J]. Hum Reprod,2005,20(7):1814-1819.
[54]Luisi S,Florio P,Reis FM,et al.Inhibins in female and male reproductive physiology:role in gametogenesis,cnception,implantation and early pregnancy[J].Hum Reprod Update,2005,11(2):123-135
[55]王谨,程忠平,谢彦等。抑制素B与多囊卵巢综合征的相关性研究。中国临床医学,2009,16(2)：256-257
[56]Willis DS,Watson H,Mason HD,et al.Premature response to luteinizing hormone of granulosa cells from anovulatory women with polycystic ovary syndrome:relevance to mechanism of anovulation[J].J Clin Endocrinol Metab,1998,83(11):3984-3991
[57]曾海涛,梁晓燕,姚书忠,等,多囊卵巢综合征卵泡颗粒细胞提前对黄体生成素反应[J].中国病理生理杂志,2007,23(2)：3732-2375
[58]Tilly JL,Kowalski KI,Schomberg DW,et al.Apoptosis in atretic ovarian follicles is associated with selective decreases in messenger ribonucleic acid transcripts for gonadotropin receptors and cytochrome P450 aromatase[J]. Endocrinology,1992,131 (4); 1670-1676
[59]Tarlatzis BC,Zepiridis L,Grimbizis G,et al.Clinical management of low ovarian response to stimulation for IVF:a systematic review.Hum Reprod Update.2003,9(1):61-76
[60]Barbieri RL,Sluss PM,Powers RD,et al.Association of body mass index,age,and cigarette smoking with serum testosterone levels in cycling women undergoing in vitro fertiliczation.Fertil steril,2005,83 (2):302-308
[61]楼航英,黄荷凤。超排卵时卵巢对促性腺激素的反应与颗粒细胞促卵泡激素受体的表达。浙江大学学报,2002,31(3)：151-154]。
[62]蔡婕,高惠娟,黄荷凤。超排卵周期卵巢的反应性与卵泡颗粒细胞中卵泡刺激素受体表达的关系。中华妇产科杂志,2006,41(6)：372-374。
[63]Oliveira JB,Mauri AL,Petersen CG,et al.Recombinant luteinizing hormone supplementation to recombinant follicle-stimulation hormone during induced ovarian stimulation in the GnRH-agonist protocol:A meta-analysis.J Assist Reprod Genet,2007;24(2-3):67-75
[64]Durnerin CL,Erb K,Fleming R,et al.Effects of recombinant LH treatment on folliculogenesis and responsiveness to FSH stimulation.Hum Reprod,2008,23(2):421-426
[65]Mannaerts BM,Geurts TB,Odink J.A randomized three-way cross-over study in healthy pituitary-suppressed women to compare the bioavailability of human chorionic gonadotrophin(Pregnyl) after intramuscular and subcutaneous administration.Hum Reprod.1998,13(6):1461-1464
[66]Casson,P.R.Carson S.A.,Buster J.E.et al.Replacement dehydroepiandrosterone in elderly rationale and prospects for the future.Endocrinologist,1998a,8,187-94
[67]Haning RV Jr,Hackett RJ,Flood CA,et al.Plasma dehydroepiandrosterone sulfate serves as a prehormone for 48%of follicular fluid testeosterone during treatment with menotropins J Clin Endocrinol Metab.1993;76(5):1301-1307
[68]Casson PR,Santoro N,Elkind-Hirsch K,et al.Postmenopausal dehydroepiandrosterone administration increases free insulin-like growth factor-I and decreases high-density lipoprotein:a six-month trial.Fertil steril.1998;70(1):107-110
[69]Casson P R, Lindsay MS, Pisarska MD,et al.Dehydroepiandrosterone supplementation augments ovarian stimulation in poor responders:a case series.Hum Reprod,2000,15(10):2129-2132
[70]Barad D, Gleicher N.Effect of dehydroepiandrosterone on oocyte and embryo yields,embryo grade and cell number in IVF.Hum Reprod 2006;21(11):2845-2849
[71]Gleicher N,Ryan E,Weghofer A,et al. Miscarriage rates after dehydroepiandrosterone (DHEA) supplementation in women with diminished ovarian reserve:a case control study.Reprod Biol Endocrinol,2009,7:108
[72]Fabregues F,Penarrubia J,Creus M.et al.Transdermal testosterone may improve ovarian response to gonadotrophins in low-responder IVF patient:a randomized,clinical trial.Hum Reprod,2009,24(2):349-359
[73]Balasch J, Fabregues F,Penarrubia J,et al.Pretreatment with transdermal testosterone may improve ovarian response to gonadotrophins in poor-responder IVF patients with normal basal concentrations of FSH.Hum Reprod,2006,21(7):1884-1893
[74]Massin N.Cedrin-Durnerin I,Coussieu C,et al.Effects of transdermal testosterone application on the ovarian response to FSH in poor responders undergoing assisted reproduction technique-a prospective,randomized double-blind study.Hum Reprod,2006,21(5):1204-1211
[75]Mitwally MF,Casper RF.Aromatase inhibition improves ovarian response to follicle-stimulating hormone in poor responders.Fertil Steril,2002,77(4):776-780
[76]Goswami SK,Das T,Chattopadhyay R,et al.A randomized single-blind controlled trial of letrozole as a low-cost IVF protocol in women with poor ovarian response:a preliminary report.Hum Reprod 2004,19(9):2031-2035
[77]Lossl K,Andersen AN,Loft A,et al.Androgen priming using aromatase inhibitor and hCG during early-follicular-phase GnRH antagonist down-regulation in modified antagonist protocols[J].Hum Reprod,2006;21(10):2593-2600
[78]Lossl K,Andersen CY,Loft A,et al.Short-term androgen priming by use of aromatase inhibitor and hCG before controlled ovarian stimulation for IVF.A randomized controlled trial.Hum Reprod,2008,23(8):1820-1829
[79]Biljan MM.Hemmings R and Brassard N.The outcome of 150 babies following the treatment with letrozole or letrozole and gonadotropins,Fertil Steril.2005;84(suppl 1):S64-S65
[80]Okon MA,Laird SM,Tuckerman EM,et al.Serum androgen levels in women who have recurrent miscarriages and their correlation with markers of endometrial function.Fertil Steril,1998,69(4):682-690
[2]Cedars MI,Surey E,Hamilton F,et al.Leuprolide acetate lowers circulating bioactive luteinizing hormone and testosterone concentrations during ovarian stimulation for oocyte retrieval.Fertil Steril.1990;53(4):627-631
[3]Fanchin R,de Ziegler D,Castracane VD,et al.Physiopathology of premature progesterone elevation.Fertil Steril 1995;64(4):796-801
[4]Orvieto R,Yulzari-Roll V,La Marca A,et al.Serum androgen levels in patients undergoing controlled ovarian hyperstimulation for in vitro fertilization cycles.Assisted Repred Technology.2005,21(4):218-222
[5]Andersen CY,Ziebe S.Serum levels of free androstenedione,testosterone and oestradiol are lower in the follicular phase of conceptional than of non-conceptional cycles after ovarian stimulation with a gonadotrophin-rel easing hormone agonist protocol.Hum Reprod.1992,7(10):1365-1370
[6]Check JH,Nazari A,Dietterich C.Comparison of androgen levels in conception vs.non-conception cycles following controlled ovarian stimulation using the luteal phase gonadotropin-releasing hormone agonist protocol. Gynecol Endocrinol.1995;9(3):209-214
[7]Frattarelli JL,Gerber MD.Basal and cycle androgen levels correlate with in vitro fertilization stimulation parameters but do not predict pregnancy outcome.Fertil Steril,2006,86(1):51-57
[8]Frattarelli JL,Peterson EH.Effect of androgen levels on in vitro f ertilization cycles.Fertility Sterility 2004,81 (6):1713-1714
[9]Tanbo T,Dale PO,Abyholm T.Recombinant follicle-stimulating hormone stimulates ovarian androgen synthesis in down-regulated ovulatory women.Gynecol Edocrinol.2001;15(6):407-412
[10]Tarlatzis BC,Zepiridis L,Grimbizis G,et al.Clinical management of low ovarian response to stimulation for ⅣF:a systematic review.Hum Reprod Update.2003,9(1):61-76
[1]Salas C,Julio-Pieper M,Valladares M, et al.Nerve growth factor-dependent activation of takA receptors in the human ovary results in synthesis of follicle-stimulating hormone receptors and estrogen secretion.J Clinic Endocrino,2006,91 (6):2396-2403.
[2]Tarlatzis BC,Zepiridis L,Grimbizis G,et al.Clinical management of low ovarian response to stimulation for IVF:systematic review[J].Hum Reprod Update,2003,9(1):61-76
[3]O'Shaughenssy PJ,Dudley K,Rajapaksha WR.Expression of follicle stimulating hormone-receptor mRNA during gonadal development.Mol Cell Endocrinol,2004; 125(1-2_:169-175
[4]Bogovich K.Obligatory roles for follicle-stimulating hormone(FSH),estradiol and androgens in the induction of small pilyfollicular ovarian cysts in hypophysectomized immature rats.Endocrine,2007,31 (2):179-192
[5]Scott RT,Opsahl MS,Leonardi MR,et al.Life table analysis of pregnancy rates in a general infertility population relative to ovarian reserve and patient age.Hum Reprod 1995;10(7):1706-1710
[6]Huhtaniemi IT.LH and FSH receptor mutations and their effects on puberty.Horm Res,2002,57(suppl 2):35-38
[7]Abel MH,Wootton AN,Wilkins V,et al.The effect of a null mutation in the follicle-stimulating hormone receptor gene on mouse reprodution.Endocrinology,2003,141 (5):1795-1803
[8]Pellicer A,Ballester MJ,Serrano MD,et al.Aetiological factors involved in the low response to gonadotrophins in infertile women with normal basal. serum follicle stimulating hormone levels.Hum Reprod 1994;9(5):806-811
[9]Abdennebi L,Monget P,Pisselet C,et al.Comparative expression of luteinizing hormone and follicle-stimulating hormone receptors in ovarian follicles from high and low prolific sheep breeds.Biol Reprod,1999;60(4):845-854
[10]Loutradis D,Patsoula E,Minas V,et al.FSH receptor gene polymorphisms have a role for different ovarian response to stimulation in patients entering IVF/ICSI-ET programs.J Assist Reprod Genet,2006,23(4):177-184
[11]楼航英,黄荷凤。超排卵时卵巢对促性腺激素的反应与颗粒细胞促卵泡激素受体的表达。浙江大学学报,2002,31(3)：151-154
[12]蔡婕,高惠娟,黄荷凤。超排卵周期卵巢的反应性与卵泡颗粒细胞中卵泡刺激素受体表达的关系。中华妇产科杂志。2006,41(6)：372-374
[13]Cai J,Lou HY,Dong MY,et al.Poor ovarian response to gonadotropin stimulation associated with low expression of follicle-stimulating hormone receptor in gramulosa cells.Fertil Steril,2007;87(6):1350-1356
[14]Wang H,Andoh K,Hagiwara H,et al.Effect of adrenal and ovarian androgens on type 4 follicles unresponsive to FSH in immature mice[J].Endocrinoogy,2001,142 (11):4930-4936
[15]Vendola KA,Zhou J,Adesanya OO,et al.Androgens stimulate early stages of follicular growth in the primate ovary[J].J Clin Invest,1998,101(12):2622-2629
[16]Cardenas H,Herrick JR,Pope WF.Increased ovulation rate in gilts treated with dihydrotestosterone.Reproduction.2002,123(4):527-533
[17]Casson P R, Lindsay MS, Pisarska MD,et al.Dehydroepiandrosterone supplementation augments ovarian stimulation in poor responders:a case series.Hum Reprod,2000,15(10):2129-2132
[18]Fabregues F,Penarrubia J,Creus M,et al.Transdermal testosterone may improve ovarian response to gonadotrophins in low-responder IVF patient:a randomized,clinical trial.Hum Reprod,2009,24(2):349-359
[19]Lossl K,Andersen AN,Loft A,et al.Androgen priming using aromatase inhibitor and hCG during early-follicular-phase GnRH antagonist down-regulation in modified antagonist protocols.Human Reprod,2006,21(10):2593-2600
[20]Massin N.Cedrin-Durnerin I,Coussieu C,et al.Effects of transdermal testosterone application on the ovarian response to FSH in poor responders undergoing assisted reproduction technique-a prospective,randomized double-blind study.Hum Reprod,2006,21(5):1204-1211
[21]Weil S,Vendola K,Zhou J,et al.Androgen and follicle-stimulating hormone interactions in primate ovarian follicle development. J Clin Endocrinol Metab.1999,84(8):2951-2956
[22]Catteau-Jonard S,Jamin SP,Leclerc A,et al.Anti-Mullerian Hormone,its receptor,FSH receptor,and androgen receptor genes are overexpressed by granulosa cells from stimulated follicles in women with polycystic ovary syndrome.J Clin Endocrinol metab.2008,93(11):4456-4461
[23]Simon C,Garcia Velasco JJ,Valbuena D,et al.Increasing uterine receptivety decresasing estradiol levels during the preimplantation period in high responders with the use of a follicle-stimulating hormone step-down regimen.Fertil Steril,1998,70(2):234-239
[24]Simon C,Domingguez F,Valbuena D,et al.The role of estrogen in uterine receptivity and blastecyst implantation.Trends Endtrinol,Metab,2003,14(5):197-199
[25]Forman R,Fries N,Testart J,et al.Evidence for an adverse effect of elevated serum estradiol concentrations on embryo implantation.Fertil Steril,1988,49(1):118-122 [26]Sharara F,McClamrock HD.High estradiol levels and high oocyte yield are not detrimental to in vitro fertilization outcome.Fertil Steril,1999,72(3):401-405
[1]Lossl K, Andersen AN, Loft A,et al.Androgen priming using aromatiase inhibitor and hCG during early-follicular-phase GnRH antagonist down-regulation in modified antagonist protocols.2006,21(10):2593-2600
[2]Greisen S, Ledet T, Ovesen P.Effects of androstenedione,insulin and luteinizing hormone on steroidogenesis in human granulose luteal cells.Human Reprod,2001;16(10):2061-65
[3]C Ganeff,G Chatel,Munaut,et al. The IGF system in in-vitro human dicidualization,2009,15(1):27-38
[4]Wang HS,Chard T.The role of insulin-like growth factor-I and insulinOlike growth factor-binding protein-1 in the control of human fetal growth. J Endocrinol,1992,132(1):11-19
[5]Wang HS,Chard T.IGFs and IGF-binding proteins in the regulation of human ovarian and endometrial function J Endocrinol,1999,161(1):1-13
[6]Rajaram S,Baylink DJ,Mohan S.Insulin-like growth factor-binding proteins in serum and other biological fluids:regulation and functions.Endocr Rev,1997,18(6):801-831
[7]Jones JI,Clemmons DR.Insulin-like growth factors and their binding proteins:biological actions.Endocr Rev.1995,16(1):3-34
[8]Giudice LC.Insulin-like growth factor family in Graafian follicle development and function.J Soc Gynecol Investig 2001,8(1 Suppl Proceedings):S26-9
[9]Vendola K,Zhou J,Wang J,et al.Androgens rpomote oovyte insulin-like growth factor I expression and initiation of follicle development in the primate ovary. Biol Reprod,1999,61 (2):353-357
[10]Thomas FH,Campbell BK.Armstrong DG,et al.Effects of IGF-I bioavailability on bovine preantral follicular devlopment in vitro.Reproduction,2007,133(6):1121-1128
[11]Demeestere I,Gervy C,Centner J,et al.Effect of insulin-like growth factor-I during preantral follicular culture on steroidogenesis,in vitro oocyte maturation,and embryo development in mice.Biol Reprod,2004,70(6):1664-1669
[12]Monget P,Fabre S,Mulsant P,et al.Regulation of ovarian folliculogenesisby IGF and BMP system in domestic animals.Domest Anim Endocrinol,2002,23(1-2):139-154
[13]Zhou J,Kumar TR,Matzuk MM,et al.Insulin-like growth factor I regulates gonadotropin responsiveness in the murine ovary.Mol Endocrinol, 1997,11 (13):1924-1933
[14]Zhou H,Zhang Y.Effect of growth factors on in vitro developement of caprine preantral follicle oocytes.Anim Reprod Sci,2005,90(3-4):265-272
[15]Monniaux D,Pisselet C.Control of proliferation and differentiation of ovine granulosa cells by insulin-like growth factor-I and follicle-stimulating hormone in vitro.Biol Reprod,1992,46(1):109-119
[16]Louhio H,Hovatta O,Sjoberg J,et al.The effects of insulin,and insulin-like growth factors I and II on human ovarian follicles in long-term culture.Mol Hum Reprod,2000,6(8):694-698.
[17]Hemamalini NC,Rao BS,Tamilmani G,et al.Influence of transforming growth factor-α,insulin-like growth factor-Ⅱ,epidermal growth factor or follicle stimulating hormone on in vitro development of preantral follicles in sheep.Smal Rum Res 2003;50:11-22
[18]Rajarajan K,Rao BS,Vagdevi R,et al.Effect of arious growth factors on the in vitro development of goat preantral follicles.Small Rum Res 2006;63:204-212
[19]Willis DS,Mason HD,Watson H,et al.Developmentally regulated responses of human granulosa cells to insulin-like growth factors (IGFS):IGF-I and IGF-II action mediated via the type-I IGF receptor.J Clin Endocrinol Metab,199883(4):1256-1259
[20]el-Roeiy A,Chen X.Roberts VJ,et al.Expression of insulin-like growth factor-I(IGF-I) and IGF-II and the IGF-I,IGF-Ⅱ and insulin receptor genes and localization of the gene products in the human ovary.J Clin Endocrinol Metab,1993,77(5):1411-1418
[21]Voutilainen R,Franks S,Mason HD,et al.Expression of insulin-like growth factor(IGF),IGF-binding protein and IGF receptor messenger ribonucleic acids in normal and polycystic ovaries.J Clin Endocrinol Metab.1996,81(3):1003-1008
[22]Giudice LC.The insulin-like growth factor system in normal and abnormal human ovarian follicle development.Am J Med.1995,98(1):48S-54S
[23]Jian Zhou,Carolyn B.Anatomy of th human ovarian insulin-like growth factor system.Bioloby of reproduction.1993,48,467-482.
[24]Oliver JE,Aitman JR,Powell JF,et al.IGF-I gene expression in the rat ovary is confined to the granulosa cells of developing follicles.Endocronology,1989; 124(6):2671-2679
[25]Zhou J,Chin E,Bondy C.Cellular patern of IGF-I and IGF-I receptor gene expression in the developing and mature follicle.Endocrinology, 1991:129(6):3281-3288
[26]Chang SY,Tsai MY,Huang FJ,et al.Expression of insulin-like growth factor(IGF),IGF receptor,and IGF-Binding protein messenger ribonucleic acids in luteinized granulosa cells from different size follicles after controlled ovarian hyperstimulatin.J Assist Reprod Genet,2002,19(3):1221-126
[27]Jesionowska H,Hemmings R,Guyda HJ,et al.Determination of insulin and insulin-like growth factors in the ovarian circulation.Fertil Steril,1990,53(1):88-91
[28]Chang SY,Hsieh KC,Wang HS,et al.Follicular fluid levels of insulin-like growth factor I,insulin-like growth factor binding protein I,and ovarian steroids collected during ovum pick-up.Fertil Steril,1994,62(1):1162-1167
[29]Pellegrini S,Fuzzi B.Pratesi S,et al.In-vivo stydies on ovarian insulin-like growth factor I concentrations in human preovulatory follicles and human ovarian circulation.Hum Reprod,1995,10(6):1341-1345
[30]Thierry van Dessel HJ,Chandrasekher Y,Yap OW,et al.Serum and follicular fluid levels of insulin-like growth factor I(IGF-I),IGF-II,and IGF-binding protein-1 and-3 during the normal menstrual cycle.J Clin Endocrinol Metab,1996,81(3):1224-1231
[31]Kubota T,Kamada S,Ohara M,et al.Insulin-like growth factor Ⅱ in follicular fluid of the patients with in vitro fertilization and embryo transfer.Fertil Steril,1993,59(4):844-849
[32]Hickey TE,Marrocco DL,Amato F,et al.Androgens augment the mitogenic effects of oocyte-secreted factors and growth differentiation factor 9 on porcine granulosa cells.Biol Reprod,2005,73(4):825-832
[33]Hickey TE,Marrocco DL,Gilchrist RB,et al.Interactions between androgen and growth factors in granulosa cell subtypes of porcine antral follicles.Biol Reprod,2004:71(1):45-52
[34]el-Roeiy A,Chen X,Roberts VJ,et al.Expression of the genes encoding the insulin-like growth factors(IGF-I and II),the IGF and insulin receptors,and IGF-binding proteins 1-6 and the localization of their gene products in normal and polycystic ovary syndrome ovaries.J Clin Endocrinol Metab,1994,78(6):1488-1496
[35]Ramasharma K,Cabrera CM,Li CH.Identification of insulin-like growth factor-II in human seminal and follicular fluids.Biochem Biophys Res Commun,1986,140(2):536-542
[36]Voutilainen R,Miller WL.Coordinate tropic hormone regulation of mRNAs for insulin-like growth factor Ⅱ and the cholesterol side-chain-cleavage enzyme,P450ssc[corrected],in human steroidogenic tissues.Proc Natl Acad Sci USA,1987,84(6):1590-1594.
[37]Barreca A,Del Monte P,Ponzani P,et al.Intrafollicular insulin-like trowth factor-II levels in normally ovulating women and in patients with polycystic ovary syndrome.Fertil Steril,1996,65(4):739-745.
[1]Tarlatzis BC,Zepiridis L,Grimbizis G,et al.Clinical management of low ovarian response to stimulation for IVF:systematic review[J].Hum Reprod Update,2003,9(1):61-76
[2]Hugues JN,Cedrin Durnerin IC.Revisiting gonadotrophin-releasing hormone agonist protocols and management of poor ovarian responses to gonadotrophins[J].Hum Reprod Update,1998,4(1),83-101
[3]Hofmann GE,Toner JP.Muasher SJ,et al.High-dose follicle stimulationg hormone (FSH)ovarian stimulation in low-responder patients for in vitro fertilization[J].J In Vitro Fert Embryo Transf,1989,6(5),285-289
[4]Check JH,Nowroozi K,Chase JS,et al.Ovulation induction and pregnancies in 100 consecutive women with hypergonadotropic amenorrhea[J].Ferti Steril.1990,53(5):811-6
[5]Homburg R,Eshel A,Abdalla HI,et al.Growth hormone facilitates ovulation induction by gonadotrophins.Clin Endocrionl(Oxf).1988,29(1),113-117
[6]Keay SD,Lenton EA,Cooke ID,et al.Low-dose dexamethasone augments the ovarian response to exogenous gonadotrophins leading to a reduction in cycle cancellation rate in a standard IVF programme[J].Hum Reprod,2001,16(9):1861-1865
[7]Orentreich N.Brind JL,Rizer RL,et al.Age changes and sex differences in serum dehydroepiandrosterone sulfate concentrations throughout adulthood.J Clin Endocrinol Metab.1984,59(3):551-555
[8]Frattarelli JL,Peterson EH.Effect of androgen levels on in vitro fertilization cycles[J]. Fertil Steril.2004,81(6):1713-1714
[9]Orvieto R,Yulzari-Roll V,La Marca A,et al.Serum androgen levels in patients undergoing conrolled ovarian hyperstimulation for in vitro fertilization cycles[J].Gynecol endocrinol.2005,21(4):218-222
[10]Wang H,Andoh K,Hagiwara H,et al.Effect of adrenal and ovarian androgens on type 4 follicles unresponsive to FSH in immature mice[J].Endocrinoogy 2001,142 (11):4930-4936
[11]Walters KA,Allan CM,Handelsman DJ.Androgen actions and the ovary[J].Biol Reprod,2008;78(3):380-389
[12]Vendola KA,Zhou J,Adesanya OO,et al.Androgens stimulate early stages of follicular growth in the primate ovary[J].J Clin Invest,1998,101(12):2622-2629
[13]Ware VC.The role of androgens in follicular development in the ovary.Ⅰ.A quantitative analysis of oocyte ovulation[J] J Exp Zool 1982,222(2):155-167
[14]Cardenas H,Herrick JR,Pope WF.Increased ovulation rate in gilts treated with dihydrotestosterone.Reproduction.2002,123(4):527-533
[15]Beloosesky R,Gold T,Almog B,et al.Induction of polycystic ovary by testosterone in immature female rats modulation of apoptosis and attenuation of glucose/insulin ratio.Int J Mol Med,2004,14(2):207-215
[16]Hillier SG,Whitelaw PF,Smyth CD.Follicular ostrogen synthesis:the'the two-cell,two-gonadotrophin'model revisited[J].Mol Cell Endocrinol 1994,100(1-2):51-54
[17]Quigley CA,De Bellis A,Marschke KB,et al.Androgen receptor defects:historical,clinical,and molecular perspectives.Endocr Rev,1995;16(3):271-321
[18]Hirai M,Hirata S,Osada T,et al.Androgen receptor mRNA in the rat ovary and uterus J Steroid Biochem Mol Biol.1994;49(1):1-7
[19]Tetsuka M,Hillier SGAndrogen receptor gene expression in rat granulosa cells:the role of follicle-stimulating hormone and steroid hormones[J].Endocrinology,1996,137(10):4392-4397
[20]Szoltys M,Slomczynska M.Changes in distribution of androgen receptor during maturation of rat ovarian follicles.Exp Clin Endocrinol Diabetes,2000,108(3):228-234
[21]Juengel JL,Heath DA,Quirke LD,et al.Oestrogen receptor alpha and beta,androgen receptor and progesterone receptor mRNA and protein localisation within the developing ovary and in small growing follicles of sheep[J].Reproduction,2006;131(1):81-92
[22]Hampton JH,Manikkam M,Lubahn DB,et al.Androgen receptor mRNA expression in the bovine ovary.Domest Anim Endocrinol 2004;27(1):81-88
[23]Slomczynska M,Duda M,Sl zak K.The expression of androgen receptor,cytochrome P450 aromatase and FSH receptor mRNA in the porcine ovary.Folia Histochem Cytobiol.2001;39(1):9-13.
[24]Cardenas H,Pope WF.Androgen receptor and follicle-stimulating hormone receptor in the pig ovary during the follicular phase of the estrous cycle.Mol Reprod Dev,2002;62(1):92-98
[25]Hillier SQTetsuka M,Fraser HM.Location and developmental regulation of androgen receptor in primate ovary.Hum Reprod,1997,12(1):107-111
[26]Chaffin CL,Stouffer RL,Duffy DM.Gonadotropin and steroid regulation of steroid receptor and aryl hydrocarbon receptor messenger ribonucleic acid in macaque granulosa cells during the periovulatory interval.Endocrinology,1999;140(10):4753-4760
[27]Duffy DM,Abdelgadir SE,Stott KR,et al.Androgen receptor mRNA expression in the rhesus monkey ovary.Endocrine.1999,11(1):23-30
[28]Horie k,Takakura K,Fujiwara H,et al.Immunohistochemical localization of androgen receptor in the human ovary throughout the menstrual cycle in relation to oestrogen and progesterone receptor expression.Hum Reprod 1992,7(2):184-190
[29]Weil SJ,Vendola K,Zhou J,et al.Androgen receptor gene expression in the primate ovary:cellular localization,regulation,and functional correlations.J Clin Endocrinol Metab.1998,83(7):2479-2485
[30]Catteau-Jonard S,Jamin SP,Leclerc A,et al.Anti-Mullerian Hormone,its receptor,FSH receptor,and androgen receptor genes are overexpressed by granulosa cells from stimulated follicles in women with polycystic ovary syndrome.J Clin Endocrinol metab.2008,93(11):4456-4461
[31]Weil S,Vendola K,Zhou J,et al.Androgen and follicle-stimulating hormone interactions in primate ovarian follicle development. J Clin Endocrinol Metab.1999,84(8):2951-56
[32]Yoshimura Y,Ando M,Nagamatsu S,et al.Effects of insulin-like growth factor-I on follicle growth,oocyte maturation and ovarian steroidogenesis and plasminogen activator anctivity in the rabbit,Biol Reprod,1996,55(1):152-160
[33]Zhou J,Kumar TR,Matzuk MM,et al.Insulin-like growth factor I regulates gonadotropin responsiveness in the murine ovary.Mol Endocrinol.1997;11(13):1924-1933
[34]Hickey TE,Marrocco DL,Amato F,et al.Androgens augment the mitogenic effects of oocyte-secreted factors and growth differentiation factor 9 on porcine granulosa cells.Biol Reprod,2005,73(4):825-832
[35]Hickey TE,Marrocco DL,Gilchrist RB,et al.Interactions between androgen and growth factors in granulosa cell subtypes of porcine antral follicles.Biol Reprod,2004:71(1):45-52
[36]Vendola K,Zhou J,Wang J,et al.Androgens promote oocyte insulin-like growth factor I expression and initiation of follicle development in the primate ovary.Biol Reprod,1999,61(2):353-357
[37]Hamel M,Vanselow J,Nicola ES,et al.Androstenedione increases cytochrome P450 aromatase messenger ribonucleic acid transcripts in nonluteinizing bovine granulosa cells[J].Mol Reprod Dev,2005,70(2):175-183
[38]ElBeltagy K,Honda K,Ozaki K,et al.In vitro effect of dehydroepiandrosterone sulfate on steroid receptors,aromatase,cyclooxygenase-2 expression,and steroid hormone production in preovulatory human granulose cells.Fertil Steril,2007,88(4 suppl):1135-1142
[39]Billig H,Furuta I,Hsueh AJ.Estrogens inhibit and androgens enhance ovarian Granulosa cell apoptosis.Endocrinology 1993;133 (5):2204-2212
[40]Conway BA,Mahesh VB,Mills TM.Effect of dihydrotestoterone on the growth and function of ovarian follicles in intact immature female rats primed with PMSG.J Reprod Fertil,1990;90(1):267-277
[41]Farookhi R.Effects of aromatizable and nonaromatizable androgen treatments on luteinizing hormone receptors and ovulation induction in immature rats.Boil Reprod,1985;33(2):363-369
[42]Azzolin GC,Saiduddin S.Effect of androgens on the ovarian morphology of the hypophysectomized rat.Proc Soc Exp Biol Med,1983,172(1):70-73
[43]张娟,朱桂金,王昕荣,等。多囊卵巢综合征模型鼠颗粒细胞凋亡及TRAIL蛋白的表达。中国组织化学与细胞化学杂志。2007,16(3)：352-356
[44]Zeleznik AJ,Hillier SQRoss GT.Follicle stimulating hormone-induced follicular development:an examination of the role of androgens.Biol Reprod,1979,21(3);673-681
[45]Anderiesz C,Trounson AO.The effect of testosterone on the maturation and developmental capacity of murine oocytes in vitro.Hum Reprod,1995,10(9):2377-2381
[46]Harlow CR,Shaw HJ,Hillier SG,et al.Factors influencing follicle-stimulating hormone-responsive steroidogenesis in marmoset gramulosa cell:effects of androgens and the stage of follicular maturity.Endocrinology,1988;122(6):2780-2787
[47]Zeleznik AJ,Little-lhrig L,Ramasawamy S.Administration of dihydrotestosterone to rhesus monkeys inhibits gonadotropin-stimulated ovarian steroidogenisis[J].J Clin endocrinol metab,2004,89(2);860-866
[48]孙春玲,乔杰,陈英玉。体外受精-胚胎移植中多囊卵巢综合征颗粒细胞凋亡与妊娠结局关系的探讨。中国实用妇科与产科杂志。2007,23(9)：695-697
[49]Weenen C,Laven JS,Von Bergh AR,et al.AntiMullerian homone expression pattern in the human ovary.Potential implications for initial and cyclic follicle recruitment [J],Mol Hum Reprod,2004,10(2):77-83
[50]Durlinger AL, Gruijters MJ, Kramer P, et al. Anti-Mullerian hormone attenuates the effects of FSH on follicle development in the mouse ovary[J]. Endocrinology, 2001,142(11):4891-4899.
[51]Pigny P,Merlen E,Robert Y,et al.Elevated serum level of anti-Mullerian humone in patients with polycystic ovary syndrome:relationship to the ovarian follicle excess and to the follicular arrest[J].J Clin Endocrinol Metab.2003,88(12):5957-5962
[52]Piltonen T,Morin-Papunen L,Koivunen R,et al.Serun anti-Mullerian hormone levels remain high until late reproductive age and decrease during metformin therapu in women with polycystic ovary syndrome[J].Hum Reprod,2005,20(7):1820-1826
[53]Eldar-Geva T, Margalioth EJ, Gai M, et al. Serum anti-Mullerian hormone levels during controlled ovarian hyperstimulation in women in polycystic ovaries with and without hyperandrogenism[J]. Hum Reprod,2005,20(7):1814-1819.
[54]Luisi S,Florio P,Reis FM,et al.Inhibins in female and male reproductive physiology:role in gametogenesis,cnception,implantation and early pregnancy[J].Hum Reprod Update,2005,11(2):123-135
[55]王谨,程忠平,谢彦等。抑制素B与多囊卵巢综合征的相关性研究。中国临床医学,2009,16(2)：256-257
[56]Willis DS,Watson H,Mason HD,et al.Premature response to luteinizing hormone of granulosa cells from anovulatory women with polycystic ovary syndrome:relevance to mechanism of anovulation[J].J Clin Endocrinol Metab,1998,83(11):3984-3991
[57]曾海涛,梁晓燕,姚书忠,等,多囊卵巢综合征卵泡颗粒细胞提前对黄体生成素反应[J].中国病理生理杂志,2007,23(2)：3732-2375
[58]Tilly JL,Kowalski KI,Schomberg DW,et al.Apoptosis in atretic ovarian follicles is associated with selective decreases in messenger ribonucleic acid transcripts for gonadotropin receptors and cytochrome P450 aromatase[J]. Endocrinology,1992,131 (4); 1670-1676
[59]Tarlatzis BC,Zepiridis L,Grimbizis G,et al.Clinical management of low ovarian response to stimulation for IVF:a systematic review.Hum Reprod Update.2003,9(1):61-76
[60]Barbieri RL,Sluss PM,Powers RD,et al.Association of body mass index,age,and cigarette smoking with serum testosterone levels in cycling women undergoing in vitro fertiliczation.Fertil steril,2005,83 (2):302-308
[61]楼航英,黄荷凤。超排卵时卵巢对促性腺激素的反应与颗粒细胞促卵泡激素受体的表达。浙江大学学报,2002,31(3)：151-154]。
[62]蔡婕,高惠娟,黄荷凤。超排卵周期卵巢的反应性与卵泡颗粒细胞中卵泡刺激素受体表达的关系。中华妇产科杂志,2006,41(6)：372-374。
[63]Oliveira JB,Mauri AL,Petersen CG,et al.Recombinant luteinizing hormone supplementation to recombinant follicle-stimulation hormone during induced ovarian stimulation in the GnRH-agonist protocol:A meta-analysis.J Assist Reprod Genet,2007;24(2-3):67-75
[64]Durnerin CL,Erb K,Fleming R,et al.Effects of recombinant LH treatment on folliculogenesis and responsiveness to FSH stimulation.Hum Reprod,2008,23(2):421-426
[65]Mannaerts BM,Geurts TB,Odink J.A randomized three-way cross-over study in healthy pituitary-suppressed women to compare the bioavailability of human chorionic gonadotrophin(Pregnyl) after intramuscular and subcutaneous administration.Hum Reprod.1998,13(6):1461-1464
[66]Casson,P.R.Carson S.A.,Buster J.E.et al.Replacement dehydroepiandrosterone in elderly rationale and prospects for the future.Endocrinologist,1998a,8,187-94
[67]Haning RV Jr,Hackett RJ,Flood CA,et al.Plasma dehydroepiandrosterone sulfate serves as a prehormone for 48%of follicular fluid testeosterone during treatment with menotropins J Clin Endocrinol Metab.1993;76(5):1301-1307
[68]Casson PR,Santoro N,Elkind-Hirsch K,et al.Postmenopausal dehydroepiandrosterone administration increases free insulin-like growth factor-I and decreases high-density lipoprotein:a six-month trial.Fertil steril.1998;70(1):107-110
[69]Casson P R, Lindsay MS, Pisarska MD,et al.Dehydroepiandrosterone supplementation augments ovarian stimulation in poor responders:a case series.Hum Reprod,2000,15(10):2129-2132
[70]Barad D, Gleicher N.Effect of dehydroepiandrosterone on oocyte and embryo yields,embryo grade and cell number in IVF.Hum Reprod 2006;21(11):2845-2849
[71]Gleicher N,Ryan E,Weghofer A,et al. Miscarriage rates after dehydroepiandrosterone (DHEA) supplementation in women with diminished ovarian reserve:a case control study.Reprod Biol Endocrinol,2009,7:108
[72]Fabregues F,Penarrubia J,Creus M.et al.Transdermal testosterone may improve ovarian response to gonadotrophins in low-responder IVF patient:a randomized,clinical trial.Hum Reprod,2009,24(2):349-359
[73]Balasch J, Fabregues F,Penarrubia J,et al.Pretreatment with transdermal testosterone may improve ovarian response to gonadotrophins in poor-responder IVF patients with normal basal concentrations of FSH.Hum Reprod,2006,21(7):1884-1893
[74]Massin N.Cedrin-Durnerin I,Coussieu C,et al.Effects of transdermal testosterone application on the ovarian response to FSH in poor responders undergoing assisted reproduction technique-a prospective,randomized double-blind study.Hum Reprod,2006,21(5):1204-1211
[75]Mitwally MF,Casper RF.Aromatase inhibition improves ovarian response to follicle-stimulating hormone in poor responders.Fertil Steril,2002,77(4):776-780
[76]Goswami SK,Das T,Chattopadhyay R,et al.A randomized single-blind controlled trial of letrozole as a low-cost IVF protocol in women with poor ovarian response:a preliminary report.Hum Reprod 2004,19(9):2031-2035
[77]Lossl K,Andersen AN,Loft A,et al.Androgen priming using aromatase inhibitor and hCG during early-follicular-phase GnRH antagonist down-regulation in modified antagonist protocols[J].Hum Reprod,2006;21(10):2593-2600
[78]Lossl K,Andersen CY,Loft A,et al.Short-term androgen priming by use of aromatase inhibitor and hCG before controlled ovarian stimulation for IVF.A randomized controlled trial.Hum Reprod,2008,23(8):1820-1829
[79]Biljan MM.Hemmings R and Brassard N.The outcome of 150 babies following the treatment with letrozole or letrozole and gonadotropins,Fertil Steril.2005;84(suppl 1):S64-S65
[80]Okon MA,Laird SM,Tuckerman EM,et al.Serum androgen levels in women who have recurrent miscarriages and their correlation with markers of endometrial function.Fertil Steril,1998,69(4):682-690