小鼠早期精细胞微核试验方法比较研究

Comparative Study on the Methods of Early Spermatid Micronucleus Test

将早期精细胞微核试验的胰蛋白酶分离法和机械吹打分离法两种精细胞的分离方法与 Giemsa染色法和 PAS- HE染色法两种样本染色方法分成 4个组合并对它们进行比较研究。具体做法是将昆明系健康雄鼠 2 0只 ,随机分成 4组 ,每组5只 ,第 1组为空白对照组 ,给小鼠 1次腹腔注射蒸馏水 ,第2、3和 4组为环磷酰胺组 ,分别以 80、5 5和 30 mg/kg b.w.剂量的环磷酰胺给小鼠 1次腹腔注射 ,于接毒后第 14天取材按4种组合方法制片。试验结果表明 ,4种组合的试验方法间具有很好的相关关系 ,相关系数 r在 0 .985— 0 .996之间 ;环磷酰胺在 30— 80 mg/kg b.w.剂量范围内具有剂量 -效应关系 ,效应与剂量的线性回归方程为 y=1.2 2 5 +0 .0 5 5 D。

Four methods of the early spermatid micronucleus test were investigated. The experimental Kunming male mice were randomly divided into 4 groups, 5 mice for each group. The 1 th group as control was given sterile water by single intraperitoneal injection.The 2 st , 3 nd or 4 rd group were given cyclophosphamide by single intraperitoneal injection at dosage of 80, 55 or 30 mg/kg b.w. respectively. At the 14 d after administration of drug, mice were sacrificed for slides making by the methods of trypsin isolation of spermatids and Giemsa staining, trypsin isolation of spermatids and PAS HE staining, mechanical isolation of spermatids with pipette and Giemsa staining, mechanical isolation of spermatids with pipette and PAS HE staining respectively. The results showed that the correlations of these four methods were very good. The correlation coefficient r were between 0.985—0.996. Cyclophosphamide had dose effect relation at dose of 30—80 mg/kg b.w., and its regressional equation was y=1.225+0.055D.