Key Lab of Environment and Health, School of Public Health, Xuzhou Medical University, 209 Tong-Shan Road, Xuzhou 221004, Jiangsu, China; Key Laboratory of Human Genetics and Environmental Medicine, Xuzhou Medical University, Xuzhou, Jiangsu 221004, China.
Key Lab of Environment and Health, School of Public Health, Xuzhou Medical University, 209 Tong-Shan Road, Xuzhou 221004, Jiangsu, China; Key Laboratory of Human Genetics and Environmental Medicine, Xuzhou Medical University, Xuzhou, Jiangsu 221004, China.
Reprod Toxicol. 2024 Mar;124:108552. doi: 10.1016/j.reprotox.2024.108552. Epub 2024 Jan 29.
A widely used type II pyrethroid pesticide cypermethrin (CYP) is one of endocrine disrupting chemicals (EDCs) with anti-androgenic activity to induce male reproductive toxicology. However, the mechanisms have not been fully elucidated. This study was to explore the effects of CYP on apoptosis of mouse Sertoli cells (TM4) and the roles of endoplasmic reticulum (ER)-mitochondria coupling involving 1,4,5-trisphosphate receptor type1-glucose-regulated protein 75-voltage-dependent anion channel 1 (IP3R1-GRP75-VDAC1). TM4 were cultured with different concentrations of CYP. Flow cytometry, calcium (Ca) fluorescent probe, transmission electron microscopy and confocal microscopy, and western blot were to examine apoptosis of TM4, mitochondrial Ca, ER-mitochondria coupling, and expressions of related proteins. CYP was found to increase apoptotic rates of TM4 significantly. CYP was shown to significantly increase expressions of cleaved caspase-3, cleaved poly ADP-ribose polymerase (PARP). Concentration of mitochondrial Ca was increased by CYP treatment significantly. CYP significantly enhanced ER-mitochondria coupling. CYP was shown to increase expressions of IP3R, Grp75 and VDAC1 significantly. We suggest that CYP induces apoptosis in TM4 cells by facilitating mitochondrial Ca overload regulated by ER-mitochondria coupling involving IP3R1-GRP75-VDAC1. This study identifies a novel mechanism of CYP-induced apoptosis in Sertoli cells.
一种广泛使用的 II 型拟除虫菊酯杀虫剂氯菊酯（CYP）是具有抗雄激素活性的内分泌干扰化学物质（EDCs）之一，可诱导雄性生殖毒性。然而，其机制尚未完全阐明。本研究旨在探讨 CYP 对小鼠支持细胞（TM4）凋亡的影响，以及涉及 1,4,5-三磷酸受体 1-葡萄糖调节蛋白 75-电压依赖性阴离子通道 1（IP3R1-GRP75-VDAC1）的内质网-线粒体偶联的作用。TM4 用不同浓度的 CYP 培养。流式细胞术、钙离子（Ca）荧光探针、透射电子显微镜和共聚焦显微镜以及 Western blot 用于检测 TM4 的凋亡、线粒体 Ca、内质网-线粒体偶联以及相关蛋白的表达。发现 CYP 显著增加 TM4 的凋亡率。CYP 显著增加了裂解的胱天蛋白酶-3 和裂解的多聚 ADP-核糖聚合酶（PARP）的表达。CYP 处理显著增加了线粒体 Ca 的浓度。CYP 显著增强了 ER-线粒体偶联。CYP 显著增加了 IP3R、Grp75 和 VDAC1 的表达。我们认为，CYP 通过促进涉及 IP3R1-GRP75-VDAC1 的内质网-线粒体偶联调节的线粒体 Ca 过载来诱导 TM4 细胞凋亡。本研究确定了 CYP 诱导支持细胞凋亡的新机制。
