Rotenone-induced cell apoptosis via endoplasmic reticulum stress and PERK-eIF2α-CHOP signalling pathways in TM3 cells

• Published in: Ecotoxicology and environmental safety 2024-10, Vol.284, p.116972, Article 116972
• Main Authors: Tian, Mi, Cao, Hongting, Gao, Haoxuan, Zhu, Lingqin, Wu, Yang, Li, Guanghua
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis; Apoptosis - drug effects; Cell Line; Cell Survival - drug effects; eIF-2 Kinase - metabolism; Endoplasmic reticulum stress; Endoplasmic Reticulum Stress - drug effects; Eukaryotic Initiation Factor-2 - metabolism; Insecticides - toxicity; Leydig cells; Leydig Cells - drug effects; Leydig Cells - metabolism; Male; Mice; PERK-eIF2α-CHOP pathway; Rotenone; Rotenone - toxicity; Signal Transduction - drug effects; Testis - drug effects; Testosterone; Transcription Factor CHOP - metabolism
• ISSN: 0147-6513; 1090-2414; 1090-2414
• DOI: 10.1016/j.ecoenv.2024.116972

Rotenone (ROT), a widely used natural pesticide, has an uncertain effect on reproductive toxicity. In this study, we used 20 mice distributed randomly into four groups, with each group receiving ROT doses of 0, 2, 4, and 8 mg/kg/day for 28 days. The results demonstrated that ROT induced significant testicular damage, including impaired spermatogenesis, inhibition of testosterone synthesis, and apoptosis of Leydig cells. Additionally, ROT disrupted the normal ultrastructure of the endoplasmic reticulum (ER) in testicular tissue, leading to ER stress in Leydig cells. To further explore whether ROT-induced apoptosis in Leydig cells is related to ER stress, the mouse Leydig cell line (TM3 cells) was treated with ROT at 0, 250, 500, and 1000 nM. ROT inhibited TM3 cell viability, induced cytotoxicity, and reduced testosterone content in the culture supernatants. Furthermore, ROT treatment triggered apoptosis in TM3 cells by activating ER stress and the PERK-eIF2α-CHOP signalling pathway. Pre-treatment of TM3 cells exposed to ROT with the ER stress inhibitor 4-phenylbutyric acid (4-PBA) alleviated these effects, decreasing apoptosis and preserving testosterone levels. Further intervention with the PERK inhibitor GSK2606414 reduced ROT-induced apoptosis and testosterone reduction by inhibiting PERK activity. In summary, ROT-induced male reproductive toxicity is specifically driven by apoptosis, with the PERK-eIF2α-CHOP signalling pathway activated by ER stress playing a crucial role in the apoptosis of Leydig cells triggered by ROT. [Display omitted] •ROT induced testicular toxicity in mice.•ROT exposure induced apoptosis in Leydig cells.•ROT triggered ER stress and activated the PERK-EIF2α-CHOP pathway in mouse testes and TM3 cells.•Inhibition of ER stress and the PERK pathway alleviated ROT-induced apoptosis in TM3 cells.