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

S. M. Hsia, W. Chiang, Y. H. Kuo, P. S. Wang

Research output: Contribution to journalArticle

22 Citations (Scopus)

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 μM), 8-bromo-adenosine-3′,5′-cyclic monophosphate (8-Br-cAMP, 1 mM), A23187 (10 μM), phorbol 12-myristate 13-acetate (PMA, 0.01 μM), 25-OH-cholesterol (0.1-10 μM) and pregnenolone (0.1-10 μM) 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.

Original languageEnglish
Pages (from-to)264-274
Number of pages11
JournalInternational Journal of Impotence Research
Volume18
Issue number3
DOIs
Publication statusPublished - May 2006
Externally publishedYes

Fingerprint

Coix
Granulosa Cells
Progesterone
Down-Regulation
Enzymes
1-Butanol
Endocrine System
3-Hydroxysteroid Dehydrogenases
Equine Gonadotropins
Pregnenolone
Messenger RNA
Proteins
Water
Chinese Traditional Medicine
Calcimycin
Colforsin
Chorionic Gonadotropin
Adenosine
Cytochrome P-450 Enzyme System
Chromatography

Keywords

  • 3β-HSD
  • Adlay
  • ERK1/2
  • P450scc
  • Progesterone
  • StAR

ASJC Scopus subject areas

  • Urology

Cite this

Downregulation of progesterone biosynthesis in rat granulosa cells by adlay (Coix lachryma-jobi L. var. ma-yuen Stapf.) bran extracts. / Hsia, S. M.; Chiang, W.; Kuo, Y. H.; Wang, P. S.

In: International Journal of Impotence Research, Vol. 18, No. 3, 05.2006, p. 264-274.

Research output: Contribution to journalArticle

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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 μM), 8-bromo-adenosine-3′,5′-cyclic monophosphate (8-Br-cAMP, 1 mM), A23187 (10 μM), phorbol 12-myristate 13-acetate (PMA, 0.01 μM), 25-OH-cholesterol (0.1-10 μM) and pregnenolone (0.1-10 μM) 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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