Anderson D, Dhawan A, Yu T W, Plewa M J
BIBRA International, Carshalton, Surrey, UK.
Mutat Res. 1996 Oct 1;370(3-4):159-74. doi: 10.1016/s0165-1218(96)00061-4.
The effects of the mutagens, cyclophosphamide (CP), ethyl methanesulphonate (EMS), bleomycin (BLM) and the testicular toxin ethylene glycol monomethyl ether (EGME) in bone marrow and testicular cells have been compared in the alkaline COMET assay. Sprague-Dawley rats were administered by gavage with 50, 100 and 150 mg/kg body weight (bw) of CP; 100, 200 and 300 mg/kg bw EMS; 50, 100 and 150 mg/kg bw BLM and 500, 1000 and 1500 mg/kg bw EGME. Effects were examined at week 2 after treatment for CP, EMS BLM and EGME and at weeks 5 and 6 for EGME. Bone marrow cells were removed and separated by aspiration of the femur and testicular cells by decapsulation of the testis, treating with collagenase followed by trypsin. Various statistical methods were used to analyse the data. For CP there was an increase in damage above control values for bone marrow at 50 mg/kg bw which decreased at 100 mg/kg bw, and there was mortality of the animals at 150 mg/kg bw. A similar response was found in the testicular cells. For EMS and BLM, there were only occasional slight increases in damage in bone marrow and testicular cells. Two studies were conducted with EGME. In the first, where effects were examined at week 2 after treatment, there was an increase in damage in bone marrow cells, but a larger response was observed in testicular cells. In the second study where effects were examined at weeks 5 and 6 after treatment, bone marrow and testicular cells were not affected. The overall results showed that damage persisted for 2 weeks after treatment with CP and EGME but not in weeks 5 and 6 for EGME. Various statistical methods were used to analyse the data. Statistically significant responses were produced after treatment with CP and EGME and were dose-related for EGME, but after treatment with EMS and BLM statistical increases were sporadic. These results suggest that the assay is useful for measuring DNA damage and its persistence, and for comparing the sensitivity of different target organs in vivo.
在碱性彗星试验中，比较了诱变剂环磷酰胺（CP）、甲基磺酸乙酯（EMS）、博来霉素（BLM）以及睾丸毒素乙二醇单甲醚（EGME）对骨髓细胞和睾丸细胞的影响。将斯普拉格-道利大鼠通过灌胃给予50、100和150mg/kg体重（bw）的CP；100、200和300mg/kg bw的EMS；50、100和150mg/kg bw的BLM以及500、1000和1500mg/kg bw的EGME。在给予CP、EMS、BLM和EGME治疗后第2周检查其影响，对于EGME则在第5周和第6周检查。通过抽吸股骨获取并分离骨髓细胞，通过对睾丸去包膜、用胶原酶处理后再用胰蛋白酶处理来分离睾丸细胞。使用各种统计方法分析数据。对于CP，在50mg/kg bw时骨髓损伤高于对照值，在100mg/kg bw时降低，而在150mg/kg bw时动物出现死亡。在睾丸细胞中发现了类似的反应。对于EMS和BLM，骨髓和睾丸细胞中的损伤仅偶尔有轻微增加。对EGME进行了两项研究。在第一项研究中，在治疗后第2周检查影响，骨髓细胞损伤增加，但在睾丸细胞中观察到更大的反应。在第二项研究中，在治疗后第5周和第6周检查影响，骨髓和睾丸细胞未受影响。总体结果表明，CP和EGME治疗后损伤持续2周，但EGME在第5周和第6周时损伤未持续。使用各种统计方法分析数据。CP和EGME治疗后产生了具有统计学意义的反应，且EGME的反应与剂量相关，但EMS和BLM治疗后统计学上的增加是零星的。这些结果表明，该试验可用于测量DNA损伤及其持续性，以及比较体内不同靶器官的敏感性。
