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Downregulation of progesterone biosynthesis in rat granulosa cells by adlay (Coix lachryma-jobi L. var. ma-yuen Stapf.) bran extracts

International Journal of Impotence Research volume 18, pages 264–274 (2006)Cite this article

Abstract

Adlay (Coix lachryma-jobi L. var. ma-yuen Stapf.) has long been used as a traditional Chinese medicine for dysfunctions of the endocrine system and inflammation conditions. However, the effect of adlay seed on the endocrine system has not yet been reported. In the present study, the effects and the mechanisms of methanolic extract of adlay bran (ABM) on progesterone synthesis in rat granulosa cell were studied. ABM was further partitioned with different solvents including water, 1-butanol, ethyl acetate and n-hexane. Four subfractions named ABM-Wa (water fraction), ABM-Bu (1-butanol fraction), ABM-EA (ethyl acetate fraction) and ABM-Hex (n-hexane fraction) were obtained. ABM-Bu was further fractionated using Diaion HP-20 resin column chromatography with gradient elution. Granulosa cells were prepared from pregnant mare serum gonadotropin-primed immature female rats and challenged with different reagents including human chorionic gonadotropin (hCG 0.5 IU/ml), forskolin (10 μ M), 8-bromo-adenosine-3′,5′-cyclic monophosphate (8-Br-cAMP, 1 mM), A23187 (10 μ M), phorbol 12-myristate 13-acetate (PMA, 0.01 μ M), 25-OH-cholesterol (0.1–10 μ M) and pregnenolone (0.1–10 μ M) in the presence or absence of ABM-Bu (100 μg/ml). The functions of steroidogenic enzyme including protein expression of the steroidogenic acute regulatory protein (StAR) and cytochrome P450 side-chain cleavage enzyme (P450scc) protein were investigated. Expressions of both P450scc and StAR mRNA have also been explored. We found that ABM decreased progesterone production via an inhibition on (1) the cAMP-PKA and PKC signal transduction pathway, (2) P450scc and 3β-hydroxysteroid dehydrogenase (3β-HSD) enzyme activity, (3) P450scc and StAR protein and mRNA expressions and (4) the phosphorylation of ERK1/2 in rat granulosa cells.

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Acknowledgements

The technical assistance provided by Dr Shiow-Chwen Tsai is appreciated. This study was supported by three grants (No. DOH92-TD-1002, No. DOH93-TD-1014 and DOH93-TD-F-113-053-(2)) from the Department of Health, Executive Yuan, Taiwan, ROC. The anti-P450scc antibody was kindly provided by Dr BC Chung, Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan, ROC. The anti-StAR antibody was kindly provided by Dr DM Stocco, Department of Cell Biology and Biochemistry, Texas Tech University Health Science Center, Lubbock, Texas, USA.

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  1. Graduate Institute of Food Science and Technology, National Taiwan University, Taipei, Taiwan, Republic of China

    S-M Hsia & W Chiang

  2. Department of Chemistry, National Taiwan University, Taipei, Taiwan, Republic of China

    Y-H Kuo

  3. Department of Physiology, School of Medicine, National Yang-Ming University, Taipei, Taiwan, Republic of China

    P S Wang

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Correspondence to P S Wang.

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Hsia, SM., Chiang, W., Kuo, YH. et al. Downregulation of progesterone biosynthesis in rat granulosa cells by adlay (Coix lachryma-jobi L. var. ma-yuen Stapf.) bran extracts. Int J Impot Res 18, 264–274 (2006). https://doi.org/10.1038/sj.ijir.3901405

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