磷酸三(1,3-二氯-2-丙基)酯(TDCIPP)是在各种环境介质中经常检测到的有机磷阻燃剂(OPFRs),并被证明是生殖毒性。然而,其对生精细胞的不良影响尚不清楚。在这项研究中,选择小鼠精母细胞GC-2spd(GC-2)细胞作为体外模型,以及线粒体结构和功能的影响,内质网(ER)应激,细胞凋亡及其相关分子机制进行了研究。我们的研究表明,在82.8μM的半致死浓度(LC50)的TDCIPP治疗后,细胞活力以剂量依赖性方式显着降低。50.0μM和39.6μM持续24小时,48h和72h,分别。通过膜联蛋白V-FITC/PI染色观察到细胞凋亡。此外,线粒体结构的碎片化,线粒体膜电位(MMP)的增加,减少细胞三磷酸腺苷(ATP)含量,细胞色素c的释放以及Caspase-3和Caspase-9活性的激活表明Caspase-3依赖的线粒体途径可能在GC-2细胞凋亡过程中起关键作用。此外,ER应激诱导被ER的形态改变和ER靶向基因的上调所证实,包括(Bip,eIF2α,ATF4,XBP1,CHOP,ATF6和Caspase-12)。一起来看,这些结果表明,线粒体凋亡途径和ER应激凋亡途径可能在TDCIPP诱导的GC-2细胞凋亡过程中起重要作用。因此,TDCIPP的潜在生殖毒性不容忽视.
Tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) is a frequently detected organophosphorus flame retardants (OPFRs) in various environmental media, and has been evidenced as reproductive toxicity. However, its adverse effects on spermatogenic cells are unknown. In this study, mouse spermatocyte GC-2spd (GC-2) cells were selected as an in vitro model, and the impact of mitochondrial structure and function, endoplasmic reticulum (ER) stress, cell apoptosis and the related molecular mechanisms were investigated. Our study indicated that cell viability was decreased significantly in a dose-dependent manner after TDCIPP treatment with the half lethal concentration (LC50) at 82.8 μM, 50.0 μM and 39.6 μM for 24 h, 48 h and 72 h, respectively. An apoptosis was observed by Annexin V-FITC/PI stain. In addition, fragmentation of mitochondrial structure, an increase of mitochondrial membrane potential (MMP), reduction of cellular adenosine triphosphate (ATP) content, release of cytochrome c and activation of Caspase-3 and Caspase-9 activity implicated that Caspase-3 dependent mitochondrial pathway might play a key role in the process of GC-2 cell apoptosis. Furthermore, ER stress induction was convinced by altered morphology of ER and up-regulation of ER targeting genes, including (Bip, eIF2α, ATF4, XBP1, CHOP, ATF6 and Caspase-12). Taken together, these results demonstrate that both mitochondrial apoptotic pathways and ER stress apoptotic pathways might play important roles in the process of apoptosis in GC-2 cells induced by TDCIPP treatment. Therefore, the potential reproductive toxicity of TDCIPP should not be ignored.