Characterization of benzo[a]pyrene quinone-induced toxicity to primary cultured bone marrow stromal cells from DBA/2 mice: potential role of mitochondrial dysfunction.
Characterization of benzo[a]pyrene quinone-induced toxicity to primary cultured bone marrow stromal cells from DBA/2 mice: potential role of mitochondrial dysfunction.
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苯并[a]芘醌诱导的对 DBA/2 小鼠原代培养骨髓基质细胞毒性的表征：线粒体功能障碍的潜在作用。
DOI：
10.1006/taap.1995.1015
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发表时间：
1995
影响因子：
3.8
通讯作者：
Trush,MA
中科院分区：
文献类型：
--
作者：
Zhu,H;Li,Y;Trush,MA
DBA/2小鼠经口暴露于苯并[a]芘(BP)可引起血液毒性，表现为再生障碍性贫血和白血病。由于正常的造血是由骨髓基质细胞调节的，在这项研究中，我们研究了BP及其衍生代谢物，特别是苯醌类化合物对骨髓基质的毒性。不同浓度的BP-1，6-，3，6-，6，12-或7，8-苯二酚与基质细胞孵育24小时后，细胞存活率显著降低，且呈浓度依赖性，而BP或BP-7，8-二氢二醇处理的细胞存活率无明显下降。其中BP-1，6-苯二酚对基质细胞的细胞毒性最强。BP-1，6-苯二酚的细胞毒作用也呈时间依赖关系。1，2-二硫醇-3-硫酮(D3T)对细胞内还原型谷胱甘肽(GSH)含量和苯醌还原酶(QR)活性均有明显的诱导作用，且呈浓度依赖性。但D3T预处理对BP-1，6-苯二酚的毒性没有任何保护作用。此外，QR的强效阻断剂Dicumarol或GSH生物合成的特异性阻断剂丁硫氨酸亚磺胺不能增强BP-1，6-Quone对基质细胞的细胞毒作用。在超氧化物歧化酶或过氧化氢酶存在下，BP-1，6-苯二酚的细胞毒性不受影响。然而，BP-1，6-苯二酚与基质细胞孵育后，细胞内ATP含量显著减少，线粒体形态发生改变，导致细胞存活率下降。除BP-1，6-苯二酚外，其他具有细胞毒性的BP-1，6-苯二酚也显示出耗竭基质细胞内ATP水平的能力，而对基质细胞无细胞毒性的BP不会引起细胞内ATP水平的显著下降。这些观察表明，线粒体可能是BP苯二酚的潜在靶点。综上所述，上述结果表明细胞内的GSH和QR以及活性氧似乎都不参与BP苯醌诱导的间质细胞损伤，BP苯醌可能通过直接干扰线粒体能量代谢而对基质细胞产生细胞毒作用。
Oral exposure of DBA/2 mice to benzo[a]pyrene (BP) has been shown to result in hematotoxicity which is manifested as aplastic anemia and leukemia. Since normal hematopoiesis is regulated by bone marrow stromal cells, in this study we have characterized the bone marrow stromal toxicity induced by BP and BP-derived metabolites, particularly quinones. Incubation of stromal cells with various concentrations of BP-1,6-, 3,6-, 6,12-, or 7,8-quinone for 24 hr resulted in a significant decrease of cell survival in a concentration-dependent manner, while cells treated with BP or BP-7,8-dihydrodiol did not exhibit any significant loss of cell survival. Among the BP quinones examined, BP-1,6-quinone was the most cytotoxic to stromal cells. The cytotoxicity induced by BP-1,6-quinone also exhibited a time-dependent relationship. Pretreatment of stromal cells with 1,2 dithiole-3-thione (D3T) resulted in a significant induction of both cellular reduced glutathione (GSH) content and quinone reductase (QR) activity in a concentration-dependent manner. However, D3T pretreatment did not offer any protection against BP-1,6-quinone-induced toxicity. Furthermore, dicumarol, a potent inhibitor of QR, or buthionine sulfoximine, a specific inhibitor of GSH biosynthesis, did not potentiate BP-1,6-quinone-induced cytotoxicity to stromal cells. In the presence of superoxide dismutase or catalase, BP-1,6-quinone-induced cytotoxicity was not altered. However, incubation of stromal cells with BP-1,6-quinone resulted in a significant depletion of cellular ATP content and mitochondrial morphological changes, which preceded the loss of cell survival. In addition to BP-1,6-quinone, other cytotoxic BP quinones also exhibited a capacity to deplete cellular ATP level in stromal cells, while BP, which was not cytotoxic to stromal cells, did not elicit any significant decrease in cellular ATP level. These observations suggest that mitochondria may be a potential target of BP quinones. Overall, the above results indicate that neither cellular GSH and QR nor reactive oxygen species appear to be involved in BP quinone-induced stromal cell injury and that BP quinones may elicit cytotoxicity to stromal cells through directly disrupting mitochondrial energy metabolism.