Abstract
BackgroundMidazolam is a neurological drug with diverse functions, including sedation, hypnosis, decreased anxiety, anterograde amnesia, brain-mediated muscle relaxation, and anticonvulsant activity. Since it is frequently used in children and adolescents for extended periods of time, there is a risk that it may affect their pubertal development. Here, we report a potential effect of the drug on the development of Leydig cells (LCs), the testosterone (T)-producing cells in the testis.MethodsStem LCs (SLCs), isolated from adult rat testes by a magnetic-activated cell sorting technique, were induced to differentiate into LCs in vitro for 3 weeks. Midazolam (0.1–30 μM) was added to the culture medium, and the effects on LC development were assayed.ResultsMidazolam has dose-dependent effects on SLC differentiation. At low concentrations (0.1–5 μM), the drug can mildly increase SLC differentiation (increased T production), while at higher concentrations (15–30 μM), it inhibits LC development (decreased T production). T increases at lower levels may be due to upregulations of scavenger receptor class b Member 1 (SCARB1) and cytochrome P450 17A1 (CYP17A1), while T reductions at higher levels of midazolam could be due to changes in multiple steroidogenic proteins. The uneven changes in steroidogenic pathway proteins, especially reductions in CYP17A1 at high midazolam levels, also result in an accumulation of progesterone. In addition to changes in T, increases in progesterone could have additional impacts on male reproduction. The loss in steroidogenic proteins at high midazolam levels may be mediated in part by the inactivation of protein kinase B/cAMP response element-binding protein (AKT/CREB) signaling pathway.ConclusionMidazolam has the potential to affect adult Leydig cell (ALC) development at concentrations comparable with the blood serum levels in human patients. Further studies are needed to test the effects on human cells.
Highlights
Male infertility has been rising in recent decades, but the underlying cause is still unclear
Since stem Leydig cell (SLC) in the rat testis express the surface marker cluster of differentiation 90 (CD90) [39, 41], we adopted an SLC isolation procedure based on magnetic-activated cell sorting (MACS) protocol
The PE-CD90 antibody-stained cells were positively selected with a MACS system (BD ImagTM)
Summary
Male infertility has been rising in recent decades, but the underlying cause is still unclear. Testicular Leydig cells (LCs) are the main source of testosterone (T) in mammalian males, including men. Defects in LCs could result in low serum T level that might affect, in addition to reproduction, the general physiology of males, with symptoms including changes in body composition, increased fatigue, sexual dysfunction, depressed mood, decreased cognitive function, and reduced immune response [4,5,6,7]. Midazolam is a neurological drug with diverse functions, including sedation, hypnosis, decreased anxiety, anterograde amnesia, brain-mediated muscle relaxation, and anticonvulsant activity. Since it is frequently used in children and adolescents for extended periods of time, there is a risk that it may affect their pubertal development. We report a potential effect of the drug on the development of Leydig cells (LCs), the testosterone (T)-producing cells in the testis
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