Zinc oxide nanoparticles (ZnO NPs) have shown adverse health impact on the human male reproductive system, with evidence of inducing apoptosis. However, whether or not ZnO NPs could promote autophagy, and the possible role of autophagy in the progress of apoptosis, remain unclear. In the current study, in vitro and in vivo toxicological responses of ZnO NPs were explored by using a mouse model and mouse Leydig cell line. It was found that intragastrical exposure of ZnO NPs to mice for 28 days at the concentrations of 100, 200, and 400 mg/kg/day disrupted the seminiferous epithelium of the testis and decreased the sperm density in the epididymis. Furthermore, serum testosterone levels were markedly reduced. The induction of apoptosis and autophagy in the testis tissues was disclosed by up-regulating the protein levels of cleaved Caspase-8, cleaved Caspase-3, Bax, LC3-II, Atg 5, and Beclin 1, accompanied by down-regulation of Bcl 2. In vitro tests showed that ZnO NPs could induce apoptosis and autophagy with the generation of oxidative stress. Specific inhibition of autophagy pathway significantly decreased the cell viability and up-regulated the apoptosis level in mouse Leydig TM3 cells. In summary, ZnO NPs can induce apoptosis and autophagy via oxidative stress, and autophagy might play a protective role in ZnO NPs-induced apoptosis of mouse Leydig cells.

Residues of the pesticides chlorfenapyr (CFP) and emamectin benzoate (EMB) often coexist in the environment and can be accumulated in the body. To understand the impact of these two chemicals on health, we investigated their effect on the kidneys. In this study, rats were treated with CFP and/or EMB at low/medium/high doses of 1/3/9 mg/kg/day and 0.2/0.6/1.8 mg/kg/day, respectively, via oral gavage for 60 days. Kidneys and serum samples were collected and serum biochemistry and kidney histopathological changes were analyzed and examined. Kidney metabolome alterations were analyzed by using gas chromatography–mass spectrometry. The results showed that combined exposure to CFP and EMB elevated BUN levels and induced pathological damage, which presented as thinner renal tubular epithelial cells, an abnormal glomerular morphology, and an increased fibrotic area. CFP and/or EMB disrupted glutathione metabolism and carbohydrate metabolism, resulting in the alteration of kidney metabolomes and inducing oxidative stress in the cells of kidney tissues. In addition, CFP decreased ATP content and inhibited pyruvate PDH activity in the kidneys. These findings suggest that long-term exposure to CFP and EMB at environmentally relevant levels induce alterations in the renal metabolome, oxidative stress, and an insufficient energy supply, which may contribute to renal histopathological damage.

Tri-o-cresyl phosphate (TOCP) is widely used as a plasticizer, flame retardant, and lubricant additive, but has been reported to impair spermatogenesis. However, how TOCP affects spermatogenesis remains unclear. Therefore, the objective of this study is to investigate the underlying mechanism by which TOCP disrupts spermatogenesis. In order to achieve this, adult male mice were orally administered TOCP at doses of 0, 200, or 400 mg/kg for two weeks, and we found that TOCP exposure reduced the number of germ cells and decreased sperm density. Moreover, the numbers of PCNA-positive cells and phospho-histone H3 (Ser10)-positive cells in mouse testicular tissues were significantly decreased following TOCP treatment, indicating that germ cell proliferation may be impaired. In addition, TOCP did not affect the protein expression of neuropathy target esterase (NTE) in testicular tissues but markedly inhibited its enzymatic activity (by approximately 30% relative to the control level). In vitro experiments further demonstrate that TOCP suppressed cell proliferation and mitotic progression in mouse GC-1 spg cells and excessively activated endoplasmic reticulum (ER) stress. Treatment with 4-phenylbutyric acid (4-PBA), an ER stress inhibitor, partially reversed the TOCP-induced inhibition of cell proliferation and mitosis. Furthermore, TOCP inhibited NTE activity in GC-1 spg cells, and NTE knockdown produced a phenotype similar to that observed after TOCP exposure, characterized by suppressed cell proliferation and mitotic progression. Surprisingly, ER stress was not activated in GC-1 spg cells following NTE knockdown. Collectively, these findings suggest that TOCP may impair spermatogenesis by inhibiting the proliferation and mitotic progression of mouse germ cells, potentially through mechanisms involving excessive activation of ER stress or suppression of NTE activity.