体内外染色体畸变实验检测朱砂的遗传毒性

Genotoxicity Induced by Cinnabaris Using in Vivo and in Vitro Hamster Chromosome Aberration Tests

目的：用仓鼠的骨髓细胞与CHL细胞经行体内外染色体畸变实验评价朱砂的遗传毒性。方法：体内染色体畸变实验用毛足属仓鼠给予朱砂混悬液（2.0,4.0,8.0 g·kg-1）连续灌胃5 d,处死前2 h腹腔注射秋水仙素。处死后取骨髓细胞制备染色体。油镜下观察每只动物骨髓细胞的有丝分裂指数和100个中期分裂相细胞的畸变类型。CHL细胞染色体畸变实验用朱砂浸出液终质量浓度为6.3,12.5,25.0 g·L-1作用于CHL细胞,培养24,48 h,终止培养前4 h加入秋水仙素。收获细胞,制备染色体,油镜下观察CHL细胞的200个中期分裂相细胞的畸变类型。结果：（1）与空白组比较,朱砂浸出液12.5,25.0 g·L-1给药组的CHL细胞染色体畸变率均升高（P〈0.05,P〈0.01）,且有明显的量效关系。（2）仓鼠体内实验与朱砂血清给药CHL细胞的畸变率无显著性差异。结论：（1）在本实验中朱砂浸出液在12.5 g·L-1以上时能致体外CHL细胞染色体畸变,而朱砂混悬液在8.0 g·kg-1下仓鼠体内染色体畸变实验未出现阳性结果。（2）仓鼠体内染色体畸变实验可作为研究中药新药遗传毒性评价实验方法之一。

Objective： To evaluate the genotoxicity of Cinnabaris by using in vitro CHL cell and in vivo Hamster（Phodopus sungorus） chromosome aberration tests. Method： In vivo chromosome aberration test,hamsters（P. sungorus） were orally given cinnabar suspension at doses of 2. 0,4. 0,8. 0 g·kg-1 respectively for5 days. They were intraperitoneally injected with colchicine 2 hours before being put to death. Bone marrow cells were taken to prepare chromosome smears. Mitotic index of bone marrow cells and types of chromosome aberration in 100 metaphase cells per animal were observed under the microscope oil lens. In CHL cell chromosome aberration test,cinnabar extracts were used to react with cells,with the final concentration of 6. 3,12. 5,25. 0 g·L-1,and cultured for 24 or 48 h. Colchicine was added and cultured for 4 hours. The cells were collected to prepare chromosome smears. Types of chromosome aberration in 200 metaphase CHL cells per dose were observed under the microscope oil lens. Result：（1） Obvious chromosome aberrations were observed at doses of 12. 5,25. 0 g·L-1 in CHL cell chromosome aberration test（P〈0. 05, P〈0. 01）.（2） Compared with the negative control,the chromosome aberration rates in both in vivo chromosome aberration test and in vitro CHL cell chromosome aberration test after administration showed no statistically significant difference,with no dose-effect relations. Conclusion：（1） Cinnabar can cause chromosome aberrations in vitro at the dose of 12. 5 g·L-1 in this test. However,there was no positive result in the cinnabar suspension chromosomal aberration test at the dose of 8. 0 g·kg-1.（2） Hamster（P. sungorus） in vivo chromosome aberration test can be used as an experimental method for evaluating the genotoxicity of new traditional Chinese medicines.