Gentilhomme E, Neveux Y, Hua A, Thiriot C, Faure M, Thivolet J
Centre d'Etudes du Bouchet, BP no 3, 91710 Vert le Petit, France.
Toxicol In Vitro. 1992 Mar;6(2):139-47. doi: 10.1016/0887-2333(92)90007-e.
Human keratinocyte cultures were treated with bis(betachloroethyl)sulphide (BCES), an alkylating and vesicant agent. At concentrations of 5 x 10(-4) to 5 x 10(-3)m, spontaneous detachment of the epithelium from the culture plate was observed, reproducing in vitro the cutaneous vesication observed in vivo. Progressive cellular alterations were shown with increasing concentrations of BCES (5 x 10(-5) to 5 x 10(-3)m). At low concentrations (5 x 10(-5)m), lesions of the nucleus, a significant target for BCES, were observed, along with lesions in the cytoplasmic organelles. An acute, dose-dependent depletion of cellular glutathione was observed, which occurred within 1 hr of treatment. Mechlorethamine, an analogue of BCES, induced at equivalent doses the same glutathione depletion and similar spontaneous detachment in vitro. We suggest that BCES, in addition to its genetic effects, acts by direct metabolic toxicity and induces glutathione depletion by direct conjugation. The lesions obtained in vitro reproduced those observed in vivo. Human keratinocyte cultures can be proposed as a good model for the study of the mechanisms of action of BCES.
用人角质形成细胞培养物用双（β-氯乙基）硫化物（BCES）处理，BCES是一种烷基化和起泡剂。在浓度为5×10⁻⁴至5×10⁻³m时，观察到上皮细胞从培养板上自发脱落，在体外重现了体内观察到的皮肤水疱形成。随着BCES浓度增加（5×10⁻⁵至5×10⁻³m），显示出渐进性细胞改变。在低浓度（5×10⁻⁵m）时，观察到细胞核（BCES的一个重要靶点）的损伤以及细胞质细胞器的损伤。观察到细胞内谷胱甘肽急性、剂量依赖性耗竭，这在处理后1小时内发生。BCES的类似物氮芥在等效剂量下在体外诱导相同的谷胱甘肽耗竭和类似的自发脱落。我们认为，BCES除了其遗传效应外，还通过直接代谢毒性起作用，并通过直接结合诱导谷胱甘肽耗竭。体外获得的损伤重现了体内观察到的损伤。人角质形成细胞培养物可被提议作为研究BCES作用机制的良好模型。
