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Influences of flavones on cell viability and cAMP-dependent steroidogenic gene regulation in MA-10 Leydig cells

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Abstract

Testicular Leydig cells are major contributors of androgen synthesis and secretion, which play an important role in testis development, normal masculinization, maintenance of spermatogenesis, and general male fertility. The rate-limiting step in testosterone biosynthesis involves the transfer of cholesterol to the mitochondrial inner membrane by the steroidogenic acute regulatory (Star) protein, a critical factor in steroid hormone biosynthesis. Once inside the mitochondria, cholesterol is metabolized by the steroidogenic enzyme Cyp11a1 to pregnenolone, which is further converted to testosterone by the action of other steroidogenic enzymes. Interestingly, the Star protein level declines during Leydig cell aging, resulting in defective mitochondrial cholesterol transfer and lower testosterone production. It is possible to delay the age-related decline in testosterone production by increasing Star and/or Cyp11a1 gene expression using supplementation with flavonoids, a group of polyphenolic compounds widely distributed in fruits and vegetables. In this study, we examined whether the distribution of hydroxyl groups among flavones could influence their potency to stimulate steroidogenesis within Leydig cells. Low levels of apigenin, luteolin, chrysin, and baicalein (10 μM) stimulated cAMP-dependent Star, Cyp11a1, and Fdx1 promoters’ activation and may increase steroidogenesis within Leydig cells. Indeed, luteolin effectively increased cAMP-dependent accumulation of progesterone from MA-10 Leydig cells, possibly through activation of Star and Fdx1 transcription. Thus, dietary luteolin could be potentially effective to maintain steroid production within aging males.

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Abbreviations

bp:

Base pair

cAMP:

Cyclic adenosine monophosphate

Cebp:

CCAAT/enhancer binding protein

Cox2:

Cyclooxygenase-2

Creb:

cAMP response element binding protein

Cyp11a1:

Cholesterol side-chain cleavage enzyme

ELISA:

Enzyme-linked immunosorbent assay

Dax1:

Dosage-sensitive sex reversal, adrenal hypoplasia critical region, on chromosome X, gene 1

Fdx1:

Ferredoxin 1

FSK:

Forskolin

LH:

Luteinizing hormone

NFkB:

Nuclear factor kappa B

PKA:

Protein kinase A

Star:

Steroidogenic acute regulatory protein

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Acknowledgements

The current work was funded by the New Brunswick Innovation Foundation (NBIF) (#IAR2015 to L.J.M.; #RIF2015-014 to M.T.) and the Natural Sciences and Engineering Research Council (NSERC) of Canada (#386557 to L.J.M.; #04560 to M.T.).

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  1. Biology Department, Université de Moncton, 18, avenue Antonine Maillet, Moncton, New Brunswick, E1A 3E9, Canada

    Michelle Cormier, Firas Ghouili, Pauline Roumaud, William Bauer & Luc J. Martin

  2. Chemistry and Biochemistry Department, Université de Moncton, Moncton, New Brunswick, E1A 3E9, Canada

    Mohamed Touaibia

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  1. Michelle Cormier
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  2. Firas Ghouili
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Correspondence to Luc J. Martin.

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Supplementary Figure 1

. Chemical structures of the flavones apigenin, luteolin, chrysin and baicalein. (PDF 33 kb)

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Cormier, M., Ghouili, F., Roumaud, P. et al. Influences of flavones on cell viability and cAMP-dependent steroidogenic gene regulation in MA-10 Leydig cells. Cell Biol Toxicol 34, 23–38 (2018). https://doi.org/10.1007/s10565-017-9395-8

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