Asano N, Hagiwara T
Toxicological Research Division, Nitto Denko Corporation, Osaka, Japan.
Mutat Res. 1992 Feb-Mar;278(2-3):153-7.
The peripheral blood micronucleus test using the acridine orange (AO) supravital staining method was validated with the potent bone marrow clastogen 2-acetylaminofluorene (2-AAF). 2-AAF induced micronuclei in peripheral blood reticulocytes dose-dependently as well as in bone marrow polychromatic erythrocytes. The incidence of micronucleated reticulocytes (MNRETs) peaked 48 h after a single treatment in both CD-1 and BDF1 mice, and the incidence of micronucleated polychromatic erythrocytes (MNPCEs) peaked 24 or 48 h after treatment. The maximum incidences of MNRETs were always higher than those of MNPCEs in both mouse strains treated once. In the double-treatment regime, the maximum incidence of MNRETs was observed at 24 h after the second treatment in each strain. The incidences of MNRETs in BDF1 mice were higher than in CD-1 mice after a single treatment but were comparable after double treatment. These results indicate that the peripheral blood micronucleus test using AO supravital staining is as sensitive as the conventional bone marrow assay. The new staining method can be performed more easily than the original smear method using either bone marrow or peripheral blood cells. Thus, the peripheral blood method using AO supravital staining is a possible alternative to the conventional bone marrow assay.
采用吖啶橙（AO）活体染色法的外周血微核试验，通过强效骨髓断裂剂2-乙酰氨基芴（2-AAF）进行了验证。2-AAF可使外周血网织红细胞及骨髓嗜多染红细胞剂量依赖性地诱导产生微核。在CD-1和BDF1小鼠中，单次处理后48小时，微核网织红细胞（MNRET）的发生率达到峰值，而微核嗜多染红细胞（MNPCE）的发生率在处理后24或48小时达到峰值。在两种小鼠品系中，单次处理后MNRET的最大发生率总是高于MNPCE。在双重处理方案中，每个品系在第二次处理后24小时观察到MNRET的最大发生率。单次处理后，BDF1小鼠中MNRET的发生率高于CD-1小鼠，但双重处理后两者相当。这些结果表明，采用AO活体染色的外周血微核试验与传统骨髓试验一样敏感。新的染色方法比使用骨髓或外周血细胞的原始涂片方法更容易操作。因此，采用AO活体染色的外周血方法可能是传统骨髓试验的一种替代方法。
