ETHANOL-INDUCED INHIBITION OF TESTOSTERONE BIOSYNTHESIS IN RAT LEYDIG CELLS: ROLE OF MITOCHONDRIAL SUBSTRATE SHU1TLES AND CITRATE

Abstract Background Leydig cells reflect the activation of inflammation, decrease of androgen production, inhibition of cell growth and promotion of cell apoptosis under orchitis. Maternally expressed gene 3 (MEG3) exerts a crucial role in various human diseases, but under orchitis, the role and underlying molecular mechanism of MEG3 in Leydig cells remain unclear. Methods Lipofectamine 2000 was used for the cell transfections. qPCR and western blots assay were applied to assess the gene expression. ELISA assay was used to measure the TNFα, IL6 and testosterone secretion. CCK8 and EdU assay was employ to test the cell viability and proliferation respectively. Luciferase reporter and RIP assay were introduced to detect the binding of miR-93-5p with MEG3 and PTEN. Results Lipopolysaccharides (LPS) induced TNFα and IL6 secretion, lowered testosterone production, inhibited cell viability and proliferation, and induced cell apoptosis in Leydig cells. MEG3 was upregulated in Leydig cells treated with LPS and that knockdown of MEG3 inhibited the role of LPS in Leydig cells. MEG3 absorbed miR-93-5p and that suppression of miR-93-5p restored the role of silenced MEG3 in Leydig cells under LPS treatment. miR-93-5p inhibited PTEN expression and that over-expressed PTEN alleviated the effect of miR-93-5p in Leydig cells treated with LPS. LPS activated the MEG3/miR-93-5p/PTEN signalling pathway in Leydig cells. Conclusions This study revealed that MEG3 serves as a molecular sponge to absorb miR-93-5p, thus leading to elevation of PTEN expression in Leydig cells under LPS treatment, offering a theoretical basis on which to establish potential new treatment strategies for orchitis.

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Sustained in vivo blockade of α1-adrenergic receptors prevented some of stress-triggered effects on steroidogenic machinery in Leydig cells

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00100.2013 ◽

2013 ◽

Vol 305 (2) ◽

pp. E194-E204 ◽

Cited By ~ 12

Author(s):

Natasa J. Stojkov ◽

Marija M. Janjic ◽

Aleksandar Z. Baburski ◽

Aleksandar I. Mihajlovic ◽

Dragana M. Drljaca ◽

...

Keyword(s):

Adrenergic Receptors ◽

Leydig Cells ◽

High Affinity ◽

Testosterone Production ◽

Transcription Profiles ◽

Steroidogenic Cells ◽

Main Regulator ◽

Stimulatory Factor

This study was designed to systematically analyze and evaluate the effects of in vivo blockade of α1-adrenergic receptors (α1-ADRs) on the stress-induced disturbance of steroidogenic machinery in Leydig cells. Parameters followed 1) steroidogenic enzymes/proteins, transcription factors, and cAMP/testosterone production; 2) the main hallmarks of stress (epinephrine, glucocorticoids); and 3) transcription profiles of ADRs and oxidases with high affinity to inactivate glucocorticoids. Results showed that sustained blockade of α1-ADRs prevented stress-induced 1) decrease of the transcripts/proteins for main steroidogenic CYPs (CYP11A1, CYP17A1); 2) decrease of Scarb1 and Hsd3b1 transcripts; 3) decrease of transcript for Nur77, one of the main activator of the steroidogenic expression; and 4) increase of Dax1 and Arr19, the main steroidogenic repressors in Leydig cells. In the same cells, the expression of steroidogenic stimulatory factor Creb1, StAR, and androgen receptor increased. In this signaling scenario, stress-induced stimulation of Adra1a/Adra1b/Adrbk1 and Hsd11b2 (the unidirectional oxidase with high affinity to inactivate glucocorticoids) was not changed. Blockade additionally stimulated stress-increased transcription of the most abundantly expressed ADRs Adra1d/Adrb1/Adrb2 in Leydig cells. In the same cells, stress-decreased testosterone production, the main marker of Leydig cells functionality, was completely prevented, while reduction of cAMP, the main regulator of androgenesis, was partially prevented. Accordingly, the presented data provide a new molecular/transcriptional base for “fight/adaptation” of steroidogenic cells and new molecular insights into the role of α1-ADRs in stress-impaired Leydig cell steroidogenesis. The results are important in term of wide use of α1-ADR selective antagonists, alone/in combination, to treat high blood pressure, nightmares associated with posttraumatic stress disorder, and disrupted sexual health.

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Polychlorinated biphenyl (Aroclor 1254) inhibits testosterone biosynthesis and antioxidant enzymes in cultured rat Leydig cells

Reproductive Toxicology ◽

10.1016/j.reprotox.2008.04.003 ◽

2008 ◽

Vol 25 (4) ◽

pp. 447-454 ◽

Cited By ~ 48

Author(s):

Palaniappan Murugesan ◽

Thirupathi Muthusamy ◽

Karundevi Balasubramanian ◽

Jagadeesan Arunakaran

Keyword(s):

Antioxidant Enzymes ◽

Leydig Cells ◽

Polychlorinated Biphenyl ◽

Aroclor 1254 ◽

Testosterone Biosynthesis

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SOMATOMEDIN-C (Sm-C) RECEPTORS IN CULTURED PIG LEYDIG CELLS (LC): CHARACTERIZATION AND REGULATION. ROLE OF Sm-C ON LC FUNCTIONS

Pediatric Research ◽

10.1203/00006450-198611000-00111 ◽

1986 ◽

Vol 20 (11) ◽

pp. 1192-1192

Author(s):

M H Perrard-Sapori ◽

P G Chatelain ◽

A Ruitton ◽

J Bertrand ◽

J M Saez

Keyword(s):

Leydig Cells ◽

Somatomedin C

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A comparative study of the action of several lutropin derivatives on rat Leydig cells

Acta Endocrinologica ◽

10.1530/acta.0.0930250 ◽

1980 ◽

Vol 93 (2) ◽

pp. 250-256 ◽

Cited By ~ 6

Author(s):

M. P. de la Llosa-Hermier ◽

C. Tertrin-Clary ◽

M. Evrard-Herouard ◽

Y. Colleaux ◽

C. Hermier ◽

...

Keyword(s):

Leydig Cells ◽

Binding Activity ◽

Camp Accumulation ◽

Lh Receptor ◽

Testosterone Production ◽

Receptor Assay ◽

Testosterone Biosynthesis ◽

Radioligand Receptor Assay ◽

Relative Potencies ◽

Good Agreement

Abstract. Rat intestinal cells prepared from testes were incubated in the presence of different lutropin derivatives obtained by chemical modification of the amino groups. The cAMP accumulation and the testosterone biosynthesis were determined in the cell homogenates. Binding determinations were carried out by a radioligand receptor assay using tritiated methylated lutropin. The binding activities — relative to native LH — of three different derivatives obtained by reductive alkylation (methylated, ethylated and isopropylated LH) were in good agreement with the relative potencies assessed by their capacity to stimulate cAMP and testosterone production. Guanidinated LH (11 — NH2 groups modified) exhibited a binding activity and a relative potency relatively high with regard to cAMP accumulation (as compared with that of native LH). Its steroidogenic potency, however, was very low. When Leydig cells were incubated in the presence of native and guanidinated LH, the testosterone production was similar to that induced by the derivative alone, indicating that the derivative exerted a competitive inhibitory action preventing the stimulation of steroidogenesis by native LH. These results suggest that a guanidinated derivative is able to bind to the LH receptor and the complex so formed is able to be coupled with an adenylate cyclase pool (or cAMP compartment) which is not connected with the steroidogenic pathway.

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