1,2-Dichloroethane (1,2-DCE) is a widely used chlorinated organic toxicant, but little is known about the cerebellar dysfunction induced by excessive exposure to it. To uncover 1,2-DCE-induced neurotoxicity in cerebellar granular cells (CGCs), and to investigate the underlying mechanisms, we explored this, both in vitro and in vivo. Our findings showed significant cell viability inhibition in human CGCs (HCGCs) treated with 1,2-DCE. Flow cytometry and mitochondrial membrane potential analyses discovered an increase in apoptotic-mediated cell death in HCGCs after 1,2-DCE treatment. This HCGC apoptosis was involved in the increases of protein expression in Cytochrome c, Caspase-3, Bad, Bim, transformation related protein 53, Caspase-8, tumor necrosis factor-α, and Survivin. Quantitative real-time PCR (qPCR) and western blot confirmed the increases in Cytochrome c, Caspase-3, cleaved Caspase-3, and Bad in HCGCs after 1,2-DCE treatment. Bax inhibitor peptide V5 rescued 1,2-DCE-induced HCGC apoptosis. Furthermore, 80 CD-1 male mice were exposed to 1,2-DCE by inhalation at 0, 100, 350, and 700 mg/m3 for 6 h/day for 4 weeks. An open field test found abnormal neurobehavioral changes in the mice exposed to 1,2-DCE. Histopathological examination showed significantly shrunken and hypereosinophilic cytoplasm with nuclear pyknosis in mouse CGCs from the 700 mg/m3 1,2-DCE group. TdT-mediated dUTP nick-end labeling assay verified significant increases in apoptotic positive cells in the mouse CGCs after 1,2-DCE exposure. We confirmed the increases in the expressions of Cytochrome c, Caspase-3, cleaved Caspase-3 and Bad in the mice exposed to 1,2-DCE. These findings suggest that 1,2-DCE exposure can induce CGC apoptosis and cerebellar dysfunction, at least in part, through mitochondrial pathway.

中文翻译：

---

1,2-二氯乙烷通过体外和体内线粒体途径诱导小脑颗粒细胞凋亡

1,2-二氯乙烷 (1,2-DCE) 是一种广泛使用的氯化有机毒物，但对过度接触它引起的小脑功能障碍知之甚少。为了揭示小脑颗粒细胞 (CGC) 中 1,2-DCE 诱导的神经毒性，并研究其潜在机制，我们在体外和体内对此进行了探索。我们的研究结果表明，在用 1,2-DCE 处理的人类 CGC (HCGC) 中存在显着的细胞活力抑制。流式细胞术和线粒体膜电位分析发现 1,2-DCE 处理后 HCGC 中凋亡介导的细胞死亡增加。这种 HCGC 细胞凋亡与细胞色素 c、Caspase-3、Bad、Bim、转化相关蛋白 53、Caspase-8、肿瘤坏死因子-α 和 Survivin 中蛋白质表达的增加有关。定量实时 PCR (qPCR) 和蛋白质印迹证实了 1,2-DCE 处理后 HCGC 中细胞色素 c、Caspase-3、裂解的 Caspase-3 和 Bad 的增加。Bax 抑制剂肽 V5 拯救了 1,2-DCE 诱导的 HCGC 细胞凋亡。此外，80 只 CD-1 雄性小鼠通过以 0、100、350 和 700 毫克/立方米的浓度吸入 1,2-DCE，持续 6 小时/天，持续 4 周。一项露天试验发现暴露于 1,2-DCE 的小鼠出现异常的神经行为变化。组织病理学检查显示，来自 700 mg/m3 1,2-DCE 组的小鼠 CGC 细胞质显着收缩和嗜酸性粒细胞增多，并伴有核固缩。TdT 介导的 dUTP 缺口末端标记试验证实，1,2-DCE 暴露后小鼠 CGC 中凋亡阳性细胞显着增加。我们证实了细胞色素 c、Caspase-3、在暴露于 1,2-DCE 的小鼠中裂解 Caspase-3 和 Bad。这些发现表明，1,2-DCE 暴露可诱导 CGC 细胞凋亡和小脑功能障碍，至少部分是通过线粒体途径。