| Summary: | 博士 === 長庚大學 === 臨床醫學研究所 === 102 === The first objective was to postulate that up-regulation of SOX9 mRNA and protein due to impairment of androgen/AR signaling in Sertoli cells might explain why high level of anti-Mullerian hormone (AMH) expression occur in these testiculopathic patients.
The another objective of this study was to determine the gene expression profiles of the androgen/androgen receptor (AR) and anti-Müllerian hormone (AMH)/ Sry-related high-mobility group box 9 (SOX9) pathways in granulosa-luteal cells from patients undergoing standard in vitro fertilization (IVF) with or without recombinant luteinizing hormone (rLH) therapy. We used biological research of testicular specimens from men with azoospermia or mouse. The ability of AR to repress the expression of SOX9 and AMH mRNA and protein were evaluated in vitro in TM4 Sertoli cells and C3H10T1/2 cells. We also determined levels of reproductive hormones in the pre-ovulatory follicular fluid and the mRNA expression levels of LHR (luteinizing hormone receptor), AR, SOX9, AMH, AR-associated protein 54 (ARA54) and ARA70 in granulosa-luteal cells by real-time reverse-transcription PCR. The effects of androgen and rLH treatments on AR and AMH expression levels were also tested in vitro using HO23 granulosa-luteal cells. We found SCOS specimens showed up-regulation of SOX9 and AMH proteins but down-regulation of AR proteins in Sertoli cells. The mRNA levels of AR were significantly lower and the SOX9, AMH mRNA levels higher in all SCOS patients compared to controls. The testosterone levels in the SCOS patients were within the normal range, but most were below the median of the controls. Furthermore, our in vitro cell line experiments demonstrated that androgen/AR signaling suppressed the gene and protein levels of AMH via repression of SOX9. In addition, we observed significant correlations between LHR and AR mRNA expression and among AR, AMH, and SOX9 mRNA expression in granulosa-luteal cells from patients undergoing standard IVF treatment. Our data show that the functional androgen/AR signaling to repress SOX9 and AMH expression is essential for Sertoli cell maturation. Impairment of androgen/AR signaling promotes SOX9-mediated AMH production, accounts for impairments of Sertoli cells in SCOS azoospermic patients. Increased expression of LHR, AR, AMH, and SOX9 is characteristic of granulosa-luteal cells from IVF/ intracytoplasmic sperm injection (ICSI) patients receiving rLH supplementation. The reciprocal relationship between AR and SOX9 demonstrated in both sertoli cells and granulosa cells with homologous embryonic developmental origin.
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