Bermudez E, Mirsalis J C, Eales H C
Environ Mutagen. 1982;4(6):667-79. doi: 10.1002/em.2860040606.
A simplified method for the quantitation of DNA damage in nonlabeled hepatocytes, using a fluorometric technique for the quantitation of DNA in conjunction with a modification of the alkaline elution technique of Kohn et al [1976], following chemical treatment in vitro and in vivo, is described. Freshly isolated hepatocytes were treated in vitro with 2-acetylaminofluorene, aflatoxin B1, and dimethylnitrosamine, then examined for DNA damage. Exposure to each of these compounds resulted in DNA damage. Hepatocytes isolated from rats treated with the hepatocarcinogens 2-acetylaminofluorene, benzidine, azoxymethane, dimethylhydrazine, dimethylnitrosamine, and diethylnitrosamine sustained DNA damage as evidenced by increased alkaline elution. DNA damage in hepatocytes was also observed as a result of treatment with methylmethanesulfonate and azaserine. The hepatotoxin carbon tetrachloride did not induce DNA damage in hepatocytes isolated from treated animals. A comparison of the induction of DNA damage and of unscheduled DNA synthesis in hepatocytes from the same animals revealed that in most cases the extent of elution of DNA from filters was proportional to the induction of DNA repair.
描述了一种用于定量未标记肝细胞中DNA损伤的简化方法，该方法在体外和体内化学处理后，结合Kohn等人[1976年]碱性洗脱技术的改进，使用荧光技术定量DNA。将新鲜分离的肝细胞在体外分别用2-乙酰氨基芴、黄曲霉毒素B1和二甲基亚硝胺处理，然后检测DNA损伤。暴露于这些化合物中的每一种都会导致DNA损伤。从用肝癌致癌物2-乙酰氨基芴、联苯胺、氧化偶氮甲烷、二甲基肼、二甲基亚硝胺和二乙基亚硝胺处理的大鼠中分离出的肝细胞持续存在DNA损伤，碱性洗脱增加证明了这一点。用甲磺酸甲酯和重氮丝氨酸处理也观察到肝细胞中的DNA损伤。肝毒素四氯化碳未在从处理过的动物中分离出的肝细胞中诱导DNA损伤。对同一动物肝细胞中DNA损伤诱导和DNA非预定合成的比较表明，在大多数情况下，DNA从滤膜上的洗脱程度与DNA修复的诱导程度成正比。
