The effect of dehydroepiandrosterone supplementation on ovarian response is associated with androgen receptor in diminished ovarian reserve women

Abstract

Background: Diminished ovarian reserve(DOR) is associated with female infertility and poor response to ovarian stimulation. Our objective was to assess the effect of dehydroepiandrosterone(DHEA) on DOR women and to explore whether the improvement of ovarian response after DHEA supplementation was dependent on the expression levels of androgen receptor(AR).

Methods: A prospective cohort study was performed in the Department of Human Reproductive Medicine, Beijing Obstetrics and Gynecology Hospital during August 2014 to August 2016. 103 DOR women who completed the study were divided into the DHEA group (n = 53), which received DHEA supplementation (25 mg three times a day) for 8 weeks, and the control group (n = 50), which did not receive DHEA, before the IVF cycles. Serum hormone levels(FSH, LH, E₂, T, DHEAs, AMH, INHB), antral follicle count(AFC) and the expression of AR and FSH receptor(FSHR) in granulosa cells(GCs) were measured, meanwhile ovarian response parameters and IVF outcomes were compared. The GCs from another 36 DOR women were cultured with different concentrations of DHEA in vitro. Then, we compared the expression of AR and FSHR in GCs according to the different numbers of oocytes retrieved both in DHEA and control group.

Results: In the present study, DHEA supplementation resulted in significantly higher levels of serum T(P = 0.047), DHEAs(P = 0.019) and AR mRNA expression in GCs(P = 0.049). In vitro experiment, the protein and mRNA expression of AR and FSHR in the preovulatory GCs were significantly increased in response to DHEA supplementation(P <0.05). No significant differences were found in ovarian reserve, ovarian response, or IVF outcomes between the two groups. Subgroup analyses showed the levels of AR and FSHR mRNA in GCs were significantly increased in DHEA group with ≥5 oocytes retrieved(P <0.05).

Conclusion: DHEA supplementation can increase the expression of AR in preovulatory GCs both in vivo and in vitro. The selective beneficial effects of DHEA supplementation on ovarian response in DOR women may depend on the increasing expression of AR and FSHR in GCs.

Trial registration: The Chinese Clinical Trial Registry ( ChiCTR-IPR-15006126 ). Retrospectively Registered 19 March 2015.

Keywords: dehydroepiandrosterone/diminished ovarian reserve/androgen receptor/follicle stimulation hormone receptor/in vitro fertilization.

Fig. 1

Quantitative analysis of AR and FSHR mRNA levels in preovulatory GCs collected at oocyte retrieval. The numbers of collected GCs were 30 in DHEA and 29 in control group. Each bar represents the mean ± SD. Concomitant detection of GADPH mRNA in the real-time RT-PCR reaction served as a reference for relative quantification. Data were the mean values ± SD and all experiments were repeated three times (* P < 0.05)

Fig. 2

DHEA up-regulates AR and FSHR expression in preovulatory GCs in vitro. Granulosa cells collected from another 36 DOR women without DHEA supplementation before IVF/ISCI were treated with DMSO (Ctrl) and different concentrations (10, 20, 30, 40, 50 ng/ml) of DHEA. mRNA (Fig. 2A.) and protein (Fig. 2b.) of AR and FSHR were collected at 24 h and 48 h after DHEA treatment and examined using RT-qPCR and Western blot respectively. Data were the mean values ± SD and all experiments were repeated three times (* P < 0.05)

Fig. 3

The DHEA group (n = 30) and the control group (n = 29) were divided into subgroups based on the number of oocytes retrieved (≥5 or <5) respectively. The expression of AR and FSHR mRNA in DHEA group with more oocytes retrieved (n = 16) were significantly increasing than those with less oocytes retrieved (n = 14) (*P < 0.05). The expression of AR and FSHR didn’t reach statistically significant difference in control group (P > 0.05), and the number of collected GCs was 12 cases in oocytes retrieved ≥5 group and 17 cases in oocytes retrieved <5 group. AR and FSHR mRNA were examined using RT-qPCR. Data were the mean values ± SD and all experiments were repeated three times