Maternal exposure to PM disrupting offspring spermatogenesis through induced sertoli cells apoptosis via inhibin B hypermethylation in mice.

Particulate Matter 2.5 (PM) disrupts endocrine functions and may negatively affect sperm quality and quantity in males; however, the long-term effects and potential mechanisms of this effect are unknown. This study aimed to investigate the epigenetic mechanism of maternal exposure to PM-induced inhibin B hypermethylation in male offspring. In this experiment design, pregnant C57BL/6 mice were treated with two doses of PM (4.8 and 43.2 mg/kg bw). The membrane control group was given a sampling membrane and the control group received nothing. Following the formation of the vaginal plug, intratracheal instillation of PM was administered every three days until delivery of the pups. To assess the effect of PM in vitro, TM4 cells, a Sertoli-like cell line, was treated with different concentrations (0, 25, 50, 100 μg/mL) of PM for 24 h. The results displayed that Sperm motility, as well as the number of adult offspring, was decreased in the PM exposed group relative to the untreated controls. Increased vacuolization was observed in the Sertoli cells of mice that were exposed to PM in utero. The levels of inhibin and testosterone were reduced and the levels of LH and FSH increased in the PM groups relative to the untreated controls. In vitro, PM resulted in cell cycle inhibition as well as increased apoptosis in TM4 cells. Moreover, PM-induced inhibin B hypermethylation and activation of the p21/Cleaved Caspase-3 pathway resulted in TM4 cell apoptosis that was rescued through the use of a DNA methylation inhibitor. Together, our data suggest that prenatal exposure to PM results in inhibin B hypermethylation and can activate the p21/Cleaved Caspase-3 pathway, resulting in Sertoli cell apoptosis, aberrant secretion of androgen binding protein, and decreased testosterone, thus resulting in the inhibition of spermatogenesis.