Elevation of antimüllerian hormone in women with polycystic ovary syndrome undergoing assisted reproduction: effect of insulin Xing Yan Liu, M.Sc., Yun.

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Presentation transcript:

Elevation of antimüllerian hormone in women with polycystic ovary syndrome undergoing assisted reproduction: effect of insulin Xing Yan Liu, M.Sc., Yun Jie Yang, M.Sc., Chuan Ling Tang, Ph.D., Kai Wang, Ph.D., Jun-Jiang Chen, Ph.D., Xiao Ming Teng, M.Sc., Ye Chun Ruan, Ph.D., Jian Zhi Yang, M.D. Fertility and Sterility Volume 111, Issue 1, Pages 157-167 (January 2019) DOI: 10.1016/j.fertnstert.2018.09.022 Copyright © 2018 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Blood antimüllerian hormone (AMH) is elevated in women with polycystic ovary syndrome (PCOS). (A) ELISA measurement of AMH levels in the blood collected on hCG injection day (before the injection) from PCOS and non-PCOS women. n is shown in each column. ***P<.001 (t test). (B–F) Correlation analysis of the blood AMH level and (B) oocytes retrieved, (C) fertilization, (D) cleavage, (E) good-quality embryos, and (F) implantation in PCOS and non-PCOS women. Spearman coefficient correlation test. Fertility and Sterility 2019 111, 157-167DOI: (10.1016/j.fertnstert.2018.09.022) Copyright © 2018 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Antimüllerian hormone (AMH) is elevated in the follicular fluid and granulosa cells in women with polycystic ovary syndrome (PCOS). (A) ELISA measurement of AMH levels in follicular fluid from non-PCOS and PCOS women. Data are presented as mean ± SEM, n indicated at each column. ***P<.001 (t test). (B–C) Quantitative polymerase chain reaction (qPCR) analysis of (B) AMH and (C) AMHR2 mRNA levels in luteinized granulosa cells freshly isolated from non-PCOS and PCOS women. β-Actin was used as the internal control for relative mRNA level measurement by qPCR. Data are presented as mean ± SEM, n indicated at each column, *P<.05; **P<.01 (t test). Fertility and Sterility 2019 111, 157-167DOI: (10.1016/j.fertnstert.2018.09.022) Copyright © 2018 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Effect of insulin on AMH expression in human luteinized granulosa cells. Data are presented as mean ± SEM. (A) ELISA measurement of insulin levels in follicular fluid from women without and with PCOS. (B) qPCR analysis of mRNA levels of insulin receptor (INSR) in cultured luteinized granulosa cells isolated from non-PCOS and PCOS women., n indicated in each column. (C, D) qPCR analysis of mRNA levels of (C) AMH and (D) AMHR2 in cultured luteinized granulosa cells isolated from non-PCOS and PCOS women, after incubation with insulin (0–100 ng/mL) for 24 hours. Cells isolated from 14 non-PCOS women and six PCOS women were pooled together for the experiments. n = 3 (number of independent experiments). *P<.05; **P<.01 (one-way analysis of variance [ANOVA] with post tests). (E) qPCR analysis of mRNA levels of AMH in non-PCOS granulosa cells pretreated with or without INSR antagonist GSK1904529A (5 μmol/L) for 8 hours before incubation with or without insulin (10 ng/mL) for 24 hours. Cells isolated from ten women were pooled together for the experiments. n = 3 (number of independent experiments). *P<.05 (one-way ANOVA). Abbreviations as in Figure 2. Fertility and Sterility 2019 111, 157-167DOI: (10.1016/j.fertnstert.2018.09.022) Copyright © 2018 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Effect of insulin and AMH on genes expression in human luteinized granulosa cells. Data are presented as mean ± SEM; n = 3 (number of independent experiments). *P<.05; **P<.001; ***P<.001 (one-way ANOVA). qPCR analysis of mRNA levels of (A) CYP19, (B) FSHR, (C) AR, (D) CYP17, (E) FLK, (F) IGF1R, and (G) LHR in luteinized granulosa cells from non-PCOS women, treated with or without insulin (10 ng/mL) and AMH (20 ng/mL) for 24 hours. β-Actin was used as the internal control for relative mRNA level measurement by means of qPCR. Cells isolated from nine women were pooled together for the experiments. (H) Western blotting for CYP19 in luteinized granulosa cells isolated from non-PCOS women, treated with or without insulin (10 ng/mL) or AMH (20 ng/mL). β-Actin was used as a loading control for Western blot. (I) Western blotting for CYP19 in luteinized granulosa cells isolated from non-PCOS women, treated with or without INSR antagonist GSK1904529A (5 μmol/L) for 8 hours before incubation with or without insulin (10 ng/mL) for 24 hours. Cells isolated from 10 women were pooled together for the experiments. Abbreviations as in Figures 2 and 3. Fertility and Sterility 2019 111, 157-167DOI: (10.1016/j.fertnstert.2018.09.022) Copyright © 2018 American Society for Reproductive Medicine Terms and Conditions