摘要:
三邻甲苯磷酸酯(tri-o-cresyl phosphate,TOCP)是一种有机磷酸酯类化合物,具神经毒性作用。研究表明星形胶质细胞是有机磷化合物(organophosphorus compounds,OPs)神经毒性作用的靶点之一。为了探讨TOCP对星形胶质细胞的毒性作用,采用大鼠C6星形胶质细胞分别经0.1、0.3、1.0和3.0 mmol·L-1 TOCP染毒处理24 h,应用MTT比色法和乳酸脱氢酶(LDH)活力分析法检测细胞活力,在电子相差显微镜下观察细胞形态,二硫代二硝基苯甲酸(DNTB)比色法测定谷胱甘肽(GSH)含量和谷胱甘肽过氧化物酶(GSH-Px)活性,流式细胞仪检测分析细胞周期。结果显示,经TOCP处理24 h后,大鼠C6星形胶质细胞存活率降低,LDH释放增加,细胞形态也发生了明显的变化。GSH含量和GSH-Px活性降低,G1期细胞数量也逐渐增加。上述结果表明,TOCP对星形胶质细胞具有毒性作用,引起氧化损伤和细胞周期阻滞。
Abstract:
Tri-ortho-cresyl phosphate (TOCP), an organophosphorus ester can cause neurotoxicity. Recent studies show that astrocytes are also an important target for many organophosphorus compounds (OPs) neurotoxicity. This study aims to investigate a possible cytotoxic effect of TOCP on astrocytes in vitro. Rat C6 astroglioal cells were exposed to 0, 0.1, 0.3, 1.0 and 3.0 m mmol·L-1 TOCP for 24 h. Then cell viability was measured by MTT assay and lactate dehydrogenase (LDH) leakage. The cell morphological change was observed using light microscopy. The content of glutathione (GSH) and the activity of glutathione peroxidase (GSH-Px) were also analyzed by 5, 5dithiobis-2-nitrobenzoic acid (DTNB) chromatometry. Cell cycle progression was determined by flow cytometry. Exposure to TOCP caused decrease in cell viability, increase in LDH leakage, morphology change, decrease in the content of GSH and the activity of GPx and the G1 phase cell cycle arrest. These results suggested that TOCP has cytotoxicity effect and cause oxidative damage and phase cell cycle arrest on astroglial cells.
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