Fancd2os Reduces Testosterone Production by Inhibiting Steroidogenic Enzymes and Promoting Cellular Apoptosis in Murine Testicular Leydig Cells

Xiang Zhai; Xin-yang Li; Yu-jing Wang; Ke-ru Qin; Jin-rui Hu; Mei-ning Li; Hai-long Wang; Rui Guo

doi:https://doi.org/10.3803/EnM.2022.1431

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자료유형: 학술저널

저자정보: Xiang Zhai (Department of Biochemistry and Molecular Biology Shanxi Medical University China) Xin-yang Li (Department of Biochemistry and Molecular Biology Shanxi Medical University China) Yu-jing Wang (Department of Biochemistry and Molecular Biology Shanxi Medical University China) Ke-ru Qin (Department of Biochemistry and Molecular Biology Shanxi Medical University China) Jin-rui Hu (Department of Biochemistry and Molecular Biology Shanxi Medical University China) Mei-ning Li (Department of Biochemistry and Molecular Biology Shanxi Medical University China) Hai-long Wang (Department of Biochemistry and Molecular Biology Shanxi Medical University China) Rui Guo (Department of Biochemistry and Molecular Biology Shanxi Medical University China)

저널정보: 대한내분비학회 Endocrinology and Metabolism Endocrinology and Metabolism Vol.37 No.3

발행연도: 2022.6

수록면: 533 - 546 (14page)

DOI: https://doi.org/10.3803/EnM.2022.1431

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Background: It is well-established that serum testosterone in men decreases with age, yet the underlying mechanism of this changeremains elusive. Methods: The expression patterns of Fancd2 opposite-strand (Fancd2os) in BALB/c male mice and testicular tissue derived celllines (GC-1, GC-2, TM3, and TM4) were assessed using real-time polymerase chain reaction (RT-PCR), Western blot and immunofluorescence. The Fancd2os-overexpressing or knockdown TM3 cells were constructed by infecting them with lentivirus particlesand were used to evaluated the function of Fancd2os. The testosterone production was measured using enzyme linked immunosorbent assay (ELISA) and the steroidogenic enzymes such as steroidogenic acute regulatory protein (StAR), P450 cholesterol sidechain cleavage (P450scc), and 3β-hydroxysteroid dehydrogenase (3β-HSD) were analysed using RT-PCR. The apoptosis of TM3cells induced by ultraviolet light or testicular tissues was detected using flow cytometry, Western blot or dUTP-biotin nick end labeling (TUNEL) assays. Pearson correlation analysis was used to assess the correlation between the Fancd2os expression and TUNELpositive staining in mouse testicular Leydig cells. Results: The Fancd2os protein was predominantly expressed in mouse testicular Leydig cells and its expression increased with age. Fancd2os overexpression inhibited testosterone levels in TM3 Leydig cells, whereas knockdown of Fancd2os elevated testosteroneproduction. Fancd2os overexpression downregulated the levels of StAR, P450scc and 3β-HSD, while Fancd2os knockdown reversed this effect. Fancd2os overexpression promoted ultraviolet light-induced apoptosis of TM3 cells. In contrast, Fancd2os knockdown restrained apoptosis in TM3 cells. In vivo assays revealed that higher Fancd2os levels and mouse age were associated with increased apoptosis in Leydig cells and decreased serum testosterone levels. Pearson correlation analysis exhibited a strong positivecorrelation between the expression of Fancd2os and TUNEL-positive staining in mouse testicular Leydig cells. Conclusion: Our findings suggest that Fancd2os regulates testosterone synthesis via both steroidogenic enzymes and the apoptoticpathway.

#Testosterone #Steroidogenic acute regulatory protein #Leydig cells #Apoptosis

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