Babich H, Markenson D F, Blau L, Stern A
Department of Biological Sciences, Stern College for Women, Yeshiva University, 245 Lexington Avenue, New York, NY, USA.
Toxicol In Vitro. 1994 Oct;8(5):1075-81. doi: 10.1016/0887-2333(94)90247-x.
The cytotoxicity of the pro-oxidant fungicide dichlone (2,3-dichloro-1,4-naphthoquinone), to the human endothelial cell line, ECV304, was evaluated. The sensitivity of these cells to dichlone was intermediate between that of human hepatoblastoma HepG2 cells (least sensitive) and that of human GMO5757 fibroblasts. The midpoint cytotoxicity values for a 24-hr exposure to dichlone was about 0.02 mm when evaluated with the neutral red, acid phosphatase, and XTT tetrazolium assays. Lactic acid dehydrogenase leakage, after a 4-hr exposure, occurred initially at 0.05 mm dichlone. As with other naphthoquinones, cellular metabolism of dichlone presumably could proceed either by a one- or a two-electron reduction reaction. The enhancement of potency of dichlone towards ECV304 cells pretreated with the glutathione-depleting agents, dl-buthionine-[S,R]-sulfoximine, 1-chloro-2,4-dinitrobenzene, and 1,3-bis(chloroethyl)-1-nitrosourea; the reduction in potency of dichlone to cells pretreated with (-)-2-oxo-4-thiazolidine carboxylic acid; the decrease in intracellular glutathione on exposure to dichlone; the subtle damage to the plasma membrane of dichlone-treated cells (as detected by the leakage of lactate dehydrogenase from these cells); and the lack of potentiation of dichlone toxicity by pretreatment with dicoumarol, are all consistent with the one-electron reduction reaction as the dominant pathway and with the subsequent generation of reactive oxygen molecules. The ECV304 cell line proved to be a useful research tool to study cytotoxic injury to endothelial cells.
评估了促氧化杀菌剂二氯萘醌（2,3-二氯-1,4-萘醌）对人内皮细胞系ECV304的细胞毒性。这些细胞对二氯萘醌的敏感性介于人肝母细胞瘤HepG2细胞（最不敏感）和人GMO5757成纤维细胞之间。用中性红、酸性磷酸酶和XTT四唑盐试验评估，暴露于二氯萘醌24小时的中点细胞毒性值约为0.02 mM。暴露4小时后，乳酸脱氢酶泄漏最初发生在二氯萘醌浓度为0.05 mM时。与其他萘醌一样，二氯萘醌的细胞代谢可能通过单电子或双电子还原反应进行。用谷胱甘肽耗竭剂dl-丁硫氨酸-[S,R]-亚砜亚胺、1-氯-2,4-二硝基苯和1,3-双（氯乙基）-1-亚硝基脲预处理ECV304细胞后，二氯萘醌的效力增强；用（-）-2-氧代-4-噻唑烷羧酸预处理细胞后，二氯萘醌的效力降低；暴露于二氯萘醌后细胞内谷胱甘肽减少；二氯萘醌处理的细胞的质膜有细微损伤（通过这些细胞中乳酸脱氢酶的泄漏检测）；以及用双香豆素预处理未增强二氯萘醌的毒性，所有这些都与单电子还原反应作为主要途径以及随后活性氧分子的产生一致。事实证明，ECV304细胞系是研究内皮细胞细胞毒性损伤的有用研究工具。
