Marczynski B, Czuppon A B, Schreiber G H, Marek W, Baur X
Research Institute of Occupational Medicine at the Ruhr-University of Bochum, Gilsingstrasse 14, 44789 Bochum, Germany.
Toxicol In Vitro. 1993 Jul;7(4):531-5. doi: 10.1016/0887-2333(93)90060-i.
The immediate effect of toluene diisocyanate (TDI) on genomic DNA of sheep and rabbit white blood cells has been investigated. Following the treatment of sheep blood by TDI, the isolated DNA was analysed by anion exchange chromatography (FPLC) and melting behaviour before and after denaturation. The results of FPLC indicated that TDI induces DNA double-strand breaks in sheep white blood cells. Following denaturation of TDI-treated DNA, the results demonstrated that at least one DNA fragment, representing about 10-20% of total DNA, is cross-linked by TDI or TDI metabolite. About 39% more of double-stranded structures are reconstituted in DNA extracted from sheep white blood cells 3 hr after incubation with TDI. In addition, chromosomal DNA from rabbit blood was investigated with the 'agarose-plugs' method by pulsed-field gel electrophoresis 2 hr after incubation with TDI, and the agarose gels were quantitatively evaluated with a laser densitometer. Further studies with prednisone were conducted to correlate these results with the phenomenon of apoptosis. The in vitro results show that TDI can induce degradation of mitochondrial DNA and large DNA fragments, which result from apoptosis, to small DNA fragments.
已研究了甲苯二异氰酸酯（TDI）对绵羊和兔白细胞基因组DNA的即时影响。用TDI处理绵羊血液后，通过阴离子交换色谱法（FPLC）分析分离的DNA以及变性前后的解链行为。FPLC结果表明，TDI可诱导绵羊白细胞中的DNA双链断裂。TDI处理的DNA变性后，结果表明，至少一个占总DNA约10%-20%的DNA片段被TDI或TDI代谢物交联。与TDI孵育3小时后，从绵羊白细胞中提取的DNA中双链结构的重构增加了约39%。此外，用“琼脂糖栓”法通过脉冲场凝胶电泳研究了与TDI孵育2小时后兔血中的染色体DNA，并用激光密度计对琼脂糖凝胶进行了定量评估。用泼尼松进行了进一步研究，以将这些结果与细胞凋亡现象相关联。体外结果表明，TDI可诱导线粒体DNA和由细胞凋亡产生的大DNA片段降解为小DNA片段。
