流式细胞仪检测小鼠骨髓嗜多染红细胞微核率方法的改进

Improvement on γ-ray induced frequency of micronucleated erythrocytes in mice bone marrow by flow cytometer

目的观察经γ射线照射后不同时相点小鼠骨髓嗜多染红细胞微核率的变化,进一步完善微核流式细胞仪自动化检测方法。方法 ICR雄性小鼠给予60Coγ射线一次性全身照射,每组4只,剂量分别为1.0、3.0和6.0 Gy,与照射后0.5、2、6和12 h取材;阴性对照组给予假照射;阳性对照组注射环磷酰胺24 h后取材。结果 1.0 Gyγ射线照射后6 h,骨髓f MNPCE显著升高(P<0.01);而3.0 Gy及6.0 Gy照射后6 h,骨髓f MNPCE显著升高(P<0.01);2种方法检测结果显示均有较好的剂量、时间依赖性,并呈显著正相关性(r=0.962,P<0.05)。结论红细胞微核流式细胞仪自动化检测方法快速、简单、灵敏、客观,完全适用于小鼠骨髓红细胞微核率的检测。

Objective To detect the frequency the change of micronucleated polychromatic erythrocytes（ f MNPCE） in mice bone marrow after iradiated with ^60Co γ-ray by flow cytometer（ FCM） and to improve automatic method of assessment of f MNPCE. Methods Male ICR mice were randomly assigned to receive the γ-ray iradiation at dose of 1. 0,3. 0,6. 0 Gy（ n = 4 for each iradiation dose）,and the bone marow was collected in 0. 5,2,6,12 h after irradiation. Another 4 mice were intraperitoneally injected with cyclophosphamide once as positive control,of which the bone marow was collected in 24 h after intoxication began and 4 treated with spurious radiation as normal control. Results f MNPCE in mice bone marow increased remarkably in 6 h after exposure to1. 0 Gy γ-ray（ P〈0. 01）,and in 6 h after exposure to 3. 0 Gy and6. 0 Gy γ-ray（ P〈0. 01）. f MNPCE was detected by both FCM and microscopy showed a significant time and dose-dependent manner,and a positive corrlation was found between the results of the two methods（ r = 0. 962,P〈0. 05）. Conclusion FCM is more convenient,easy to handle,rapid,sensitive,and it is applicable to detect the frequency of micronucleated erythrocytes in mice bone marrow.