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Chronic crude garlic-feeding modified adult male rat testicular markers: mechanisms of action.

Hammami I, Amara S, Benahmed M, El May MV, Mauduit C - Reprod. Biol. Endocrinol. (2009)

Bottom Line: Because of garlic's chemical complexity, the processing methods and yield in preparations differ in efficacy and safety.The germ cell death process induced by As might be related to a decrease in testosterone production because of the reduced expression of steroidogenic enzymes (Star, Cyp11a, Hsd3b5 and Hsd17b).In contrast, AMH, RHOX5 and CDKN1B expression was decreased while GATA4 expression was increased.

View Article: PubMed Central - HTML - PubMed

Affiliation: Unité de recherche n 01/UR/08-07, Faculté de Médecine, Tunis, Tunisie. hammamiimen@hotmail.fr

ABSTRACT

Background: Garlic or Allium sativum (As) shows therapeutic effects such as reduction of blood pressure or hypercholesterolemia but side-effects on reproductive functions remain poorly investigated. Because of garlic's chemical complexity, the processing methods and yield in preparations differ in efficacy and safety. In this context, we clarify the mechanisms of action of crushed crude garlic on testicular markers.

Methods: During one month of treatment, 24 male rats were fed 5%, 10% and 15% crude garlic.

Results: We showed that crude garlic-feeding induced apoptosis in testicular germ cells (spermatocytes and spermatids). This cell death process was characterized by increased levels of active CASP3 but not CASP6. Expression of the caspase inhibitors BIRC3 and BIRC2 was increased at all doses of As while expression of XIAP and BIRC5 was unchanged. Moreover, expression of the IAP inhibitor DIABLO was increased at doses 10% and 15% of As. The germ cell death process induced by As might be related to a decrease in testosterone production because of the reduced expression of steroidogenic enzymes (Star, Cyp11a, Hsd3b5 and Hsd17b). Evaluation of Sertoli markers showed that TUBB3 and GSTA2 expression was unchanged. In contrast, AMH, RHOX5 and CDKN1B expression was decreased while GATA4 expression was increased.

Conclusion: In summary, we showed that feeding with crude garlic inhibited Leydig steroidogenic enzyme expression and Sertoli cell markers. These alterations might induce apoptosis in testicular germ cells.

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Related in: MedlinePlus

As-induced apoptosis in germ cells (TUNEL AND CASP3 immunostaining). Testes from untreated rats (A, C) or adult rats fed with 15% As (B, D). Detection of apoptotic cells was conducted through the TUNEL approach (A, B) and cleaved CASP3 immunostaining (C, D) (magnification = ×200). Red staining reveals the TUNEL positive nuclei and brown staining reveals the CASP3 cytoplasm immunostaining.
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Figure 1: As-induced apoptosis in germ cells (TUNEL AND CASP3 immunostaining). Testes from untreated rats (A, C) or adult rats fed with 15% As (B, D). Detection of apoptotic cells was conducted through the TUNEL approach (A, B) and cleaved CASP3 immunostaining (C, D) (magnification = ×200). Red staining reveals the TUNEL positive nuclei and brown staining reveals the CASP3 cytoplasm immunostaining.

Mentions: Treatment with As induced a cell death process in the adult rat testis as shown by the TUNEL approach. In the control untreated animals, very few apoptotic germ cells were observed (Fig. 1A), whereas TUNEL-positive cells were identified in rat testis treated with 15% of As (Fig. 1B). These TUNEL-positive cells were mainly spermatocytes and spermatids (Fig. 1B). The number of apoptotic germ cells in rat testes increased after treatment with As in a dose-dependent manner (Fig. 2A). A significant (p < 0.0001) increase was observed in the rats treated with 10% and 15% of As. The cell death process induced in spermatocytes and spermatids from rats fed with As was probably an apoptotic mechanism, since an immunostaining for cleaved CASP3 was detected in these cells (Fig. 1D) while only a few stained germ cells were detected in untreated rats (Fig. 1C).


Chronic crude garlic-feeding modified adult male rat testicular markers: mechanisms of action.

Hammami I, Amara S, Benahmed M, El May MV, Mauduit C - Reprod. Biol. Endocrinol. (2009)

As-induced apoptosis in germ cells (TUNEL AND CASP3 immunostaining). Testes from untreated rats (A, C) or adult rats fed with 15% As (B, D). Detection of apoptotic cells was conducted through the TUNEL approach (A, B) and cleaved CASP3 immunostaining (C, D) (magnification = ×200). Red staining reveals the TUNEL positive nuclei and brown staining reveals the CASP3 cytoplasm immunostaining.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2710331&req=5

Figure 1: As-induced apoptosis in germ cells (TUNEL AND CASP3 immunostaining). Testes from untreated rats (A, C) or adult rats fed with 15% As (B, D). Detection of apoptotic cells was conducted through the TUNEL approach (A, B) and cleaved CASP3 immunostaining (C, D) (magnification = ×200). Red staining reveals the TUNEL positive nuclei and brown staining reveals the CASP3 cytoplasm immunostaining.
Mentions: Treatment with As induced a cell death process in the adult rat testis as shown by the TUNEL approach. In the control untreated animals, very few apoptotic germ cells were observed (Fig. 1A), whereas TUNEL-positive cells were identified in rat testis treated with 15% of As (Fig. 1B). These TUNEL-positive cells were mainly spermatocytes and spermatids (Fig. 1B). The number of apoptotic germ cells in rat testes increased after treatment with As in a dose-dependent manner (Fig. 2A). A significant (p < 0.0001) increase was observed in the rats treated with 10% and 15% of As. The cell death process induced in spermatocytes and spermatids from rats fed with As was probably an apoptotic mechanism, since an immunostaining for cleaved CASP3 was detected in these cells (Fig. 1D) while only a few stained germ cells were detected in untreated rats (Fig. 1C).

Bottom Line: Because of garlic's chemical complexity, the processing methods and yield in preparations differ in efficacy and safety.The germ cell death process induced by As might be related to a decrease in testosterone production because of the reduced expression of steroidogenic enzymes (Star, Cyp11a, Hsd3b5 and Hsd17b).In contrast, AMH, RHOX5 and CDKN1B expression was decreased while GATA4 expression was increased.

View Article: PubMed Central - HTML - PubMed

Affiliation: Unité de recherche n 01/UR/08-07, Faculté de Médecine, Tunis, Tunisie. hammamiimen@hotmail.fr

ABSTRACT

Background: Garlic or Allium sativum (As) shows therapeutic effects such as reduction of blood pressure or hypercholesterolemia but side-effects on reproductive functions remain poorly investigated. Because of garlic's chemical complexity, the processing methods and yield in preparations differ in efficacy and safety. In this context, we clarify the mechanisms of action of crushed crude garlic on testicular markers.

Methods: During one month of treatment, 24 male rats were fed 5%, 10% and 15% crude garlic.

Results: We showed that crude garlic-feeding induced apoptosis in testicular germ cells (spermatocytes and spermatids). This cell death process was characterized by increased levels of active CASP3 but not CASP6. Expression of the caspase inhibitors BIRC3 and BIRC2 was increased at all doses of As while expression of XIAP and BIRC5 was unchanged. Moreover, expression of the IAP inhibitor DIABLO was increased at doses 10% and 15% of As. The germ cell death process induced by As might be related to a decrease in testosterone production because of the reduced expression of steroidogenic enzymes (Star, Cyp11a, Hsd3b5 and Hsd17b). Evaluation of Sertoli markers showed that TUBB3 and GSTA2 expression was unchanged. In contrast, AMH, RHOX5 and CDKN1B expression was decreased while GATA4 expression was increased.

Conclusion: In summary, we showed that feeding with crude garlic inhibited Leydig steroidogenic enzyme expression and Sertoli cell markers. These alterations might induce apoptosis in testicular germ cells.

Show MeSH
Related in: MedlinePlus