Background: Daidzein is a major isoflavone in soybeans. Several in vivo studies have showed that daidzein can affect immature male testosterone production. However, whether daidzein has direct action on immature male testis is unknown.
Objective: We investigated the effects of daidzein on testosterone secretion in 3-day-old and 21-day-old mouse Leydig cells with organotypic culture model.
Materials and Methods: The testes were exposed to different concentrations (10-7 to 10-4 M) of daidzein for 72 h with medium changed every 24 h. From 72 to 75 h of culture, 100 ng/ml human chorionic gonadotropin (hCG) was added. The testosterone production was determined, and the related mechanisms of daidzein action were also evaluated by measuring the mRNA levels of steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc), and 3β-hydroxysteroid dehydrogenase (3β-HSD-1) involved in testosterone biosynthesis.
Results: The results revealed that in the presence of 100 ng/ml hCG, 10-7 to 10-5 M daidzein had no significant effect on testosterone secretion in cultured 3-day-old mouse testis. But 10-4 M daidzein significantly increased testosterone concentration (p < 0.05). Daidzein in range of studied doses had no obvious influence on testosterone production in cultured 21-day-old mouse testis. RT-PCR results showed that 10-4 M daidzein had obvious influence on the mRNA levels of StAR, P450scc and 3β-HSD-1 in cultured 3-day-old mouse testis (p < 0.05).
Conclusion: These results suggest that daidzein mainly influences neonatal mouse testis function, and the influence is partially related to the upregulation of StAR, P450scc, and 3β-HSD-1 mRNA levels.
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1. Zhang LP Cui S. Effects of daidzein on testosterone synthesis and secretion in cultured mouse Leydig cells. Asian-Aust J Anim Sci. 2009; 22:618-25.
2. Wang G Zhang X Han Z Liu Z Liu W. Effects of daidzein on body weight gain serum IGF-I level and cellular immune function in intact male piglets. Asian- Aust J Anim Sci. 2002; 15:1066-70.
3. Han D Tachibana H Yamada K. Inhibition of environmental estrogen-induced proliferation of human breast carcinoma MCF-7 cells by flavonoids. In Vitro Cell Dev Biol Anim. 2001; 37:275-82.
4. Choi J Song J Choi YM Jang DJ Kim E Kim I et al. Daidzein modulations of apolipoprotein B and fatty acid synthase mRNA expression in chick liver vary depending on dietary protein levels. Asian-Aust J Anim Sci. 2006; 19:236-44.
5. Anthony MS Clarkson TB Hughes CL Jr Morgan TM Burke GL. Soybean isoflavones improve cardiovascular risk factors without affecting the reproductive system of peripubertal rhesus monkeys. J Nutr. 1996; 126:43-50.
6. Pan L Xia X Feng Y Jiang C Cui Y Huang Y. Exposure of juvenile rats to phytoestrogen daidzein impairs erectile function in a dose-related manner at adulthood. J Androl. 2008; 29:55-62.
7. Sharpe RM Martin B Morris K Greig I McKinnell C McNeilly AS et al. Infant feeding with soy formula milk: effects on the testis and on blood testosterone levels in marmoset monkeys during the period of neonatal testicular activity. Hum Reprod. 2002; 17: 1692-703.
8. Akingbemi BT Braden TD Kemppainen BW Hancock KD Sherrill JD Cook SJ et al. Exposure to phytoestrogens in the perinatal period affects androgen secretion by testicular Leydig cells in the adult rat. Endocrinology. 2007; 148:4475-88.
9. Piotrowska K Baranowska-Bosiacka I Marchlewicz M Gutowska I Nocen I Zawislak M et al. Changes in male reproductive system and mineral metabolism induced by soy isoflavones administered to rats from prenatal life until sexual maturity. Nutrition. 2011; 27: 372-9.
10. Sherrill JD Sparks M Dennis J Mansour M Kemppainen BW Bartol FF et al. Developmental exposures of male rats to soy isoflavones impact Leydig cell differentiation. Biol Reprod. 2010; 83: 488-501.
11. Cherradi N Rossier MF Vallotton MB Timberg R Friedberg I Orly J et al. Submitochondrial distribution of three key steroidogenic proteins (steroidogenic acute regulatory protein and cytochrome P450scc and 3β-hydroxysteroid dehydrogenase isomerase enzymes) upon stimulation by intracellular calcium in adrenal glomerulosa cells. J Biol Chem. 1997; 272: 7899-907.
12. Stocco DM. Recent advances in the role of StAR. Rev Reprod. 1998; 3:82-5.
13. Walsh LP Webster DR Stocco DM. Dimethoate inhibits steroidogenesis by disrupting transcription of the steroidogenic acute regulatory (StAR) gene. J Endocrinol. 2000; 167:253-63.
14. Livera G Delbes G Pairault C Rouiller-Fabre V Habert R. Organotypic culture a powerful model for studying rat and mouse fetal testis development. Cell Tissue Res. 2006; 324:507-21.
15. Laughlin AM Welsh TH Jr Love CC Varner DD Parrish AR Forrest DW et al. In vitro culture of precision-cut testicular tissue as a novel tool for the study of responses to LH. In Vitro Cell Dev Biol Anim. 2010; 46:45-53.
16. Vergouwen RP Jacobs SG Huiskamp R Davids JA de Rooij DG. Proliferative activity of gonocytes Sertoli cells and interstitial cells during testicular development in mice. J Reprod Fertil. 1991; 93:233-43.
17. Benton L Shan LX Hardy MP. Differentiation of adult Leydig cells. J Steroid Biochem Mol Biol. 1995; 53:61-8.
18. Habert R Devif I Gangnerau MN Lecerf L. Ontogenesis of the in vitro response of rat testis to gonadotropin-releasing hormone. Mol Cell Endocrinol. 1991; 82:199-206.
19. Jin L Zhang S Burguera BG Couce ME Osamura RY Kulig E et al. Leptin and leptin receptor expression in rat and mouse pituitary cells. Endocrinology. 2000; 141:333-9.
20. Akingbemi BT Ge R Rosenfeld CS Newton LG Hardy DO Catterall JF et al. Estrogen receptor-α gene deficiency enhances androgen biosynthesis in the mouse Leydig cell. Endocrinology. 2003; 144:84-93.
21. Kwon SM Kim SI Chun DC Cho NH Chung BC Park BW et al. Development of rat prostatitis model by oral administration of isoflavone and its characteristics. Yonsei Med J. 2001; 42:395-404.
22. Murkies AL Wilcox G Davis SR. Clinical review 92: Phytoestrogens. J Clin Endocrinol Metab. 1998; 83: 297-303.
23. Tham DM Gardner CD Haskell WL. Clinical review 97: Potential health benefits of dietary phytoestrogens: a review of the clinical epidemiological and mechanistic evidence. J Clin Endocrinol Metab. 1998; 83:2223-35.
24. Lund TD Munson DJ Haldy ME Setchell KD Lephart ED Handa RJ. Equol is a novel anti-androgen that inhibits prostate growth and hormone feedback. Biol Reprod. 2004; 70:1188-95.
25. Arai Y Mori T Suzuki Y Bern HA. Long-term effects of perinatal exposure to sex steroids and diethylstilbestrol on the reproductive system of male mammals. Int Rev Cytol. 1983; 84:235-68.
26. Williams K McKinnell C Saunders PT Walker M Fisher JS Turner KJ et al. Neonatal exposure to potent and environmental oestrogens and abnormalities of the male reproductive system in the rat: evidence for importance of the androgen-oestrogen balance and assessment of the relevance to man. Hum Reprod Update. 2001; 7:236-47.
27. Lassurguere J Livera G Habert R Jegou B. Time-and dose-related effects of estradiol and diethylstilbestrol on the morphology and function of the fetal rat testis in culture. Toxicol Sci. 2003; 73:160-9.
28. Atanassova N McKinnell C Turner KJ Walker M Fisher JS Morley M et al. Comparative effects of neonatal exposure of male rats to potent and weak (environmental) estrogens on spermatogenesis at puberty and the relationship to adult testis size and fertility: evidence for stimulatory effects of low estrogen levels. Endocrinology. 2000; 141:3898-907.
29. Ebling FJP Brooks AN Cronin AS Ford H Kerr JB. Estrogenic induction of spermatogenesis in the hypogonadal mouse. Endocrinology. 2000; 141: 2861-9.
30. Kuiper GG Carlsson B Grandien K Enmark E Haggblad J Nilsson S et al. Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors α and β. Endocrinology. 1997; 138:863-70.
31. Kuiper GG Lemmen JG Carlsson B Corton JC Safe SH van der Saag PT et al. Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor β. Endocrinology. 1998; 139:4252-63.
32. Delbes G Duquenne C Szenker J Taccoen J Habert R Levacher C. Developmental changes in testicular sensitivity to estrogens throughout fetal and neonatal Life. Toxicol Sci. 2007; 99:234-43.
33. Hilscherova K Jones PD Gracia T Newsted JL Zhang X Sanderson JT et al. Assessment of the effects of chemicals on the expression of ten steroidogenic genes in the H295R cell line using real-time PCR. Toxicol Sci. 2004; 81:78-89.
34. Sanderson JT. The steroid hormone biosynthesis pathway as a target for endocrine-disrupting chemicals. Toxicol Sci. 2006; 94:3-21.
35. Stocco DM Clark BJ. Regulation of the acute production of steroids in steroidogenic cells. Endocr Rev. 1996; 17:221-44.
36. Payne AH Hales DB. Overview of steroidogenic enzymes in the pathway from cholesterol to active steroid hormones. Endocr Rev. 2004; 25:947-70.