摘要
目的：研究秦皮水煎剂的遗传毒性。方法：采用小鼠淋巴瘤细胞试验（MLA）和小鼠骨髓微核试验（MNT）。MLA试验中，秦皮水煎剂（含生药）1.71，3.42，6.83，13.65 g·L-1 4个质量浓度组，超纯水阴性对照组及甲基甲烷磺酸酯、环磷酰胺阳性对照组，分别在非代谢活化（-S9）和代谢活化（＋S9）条件下与L5178Y细胞作用3 h，表达2 d，制备基因突变频率平板并培养12～13 d，计数含大、小突变细胞集落的孔数，计算每组总突变率和小集落突变百分率；MNT试验中，3个剂量组（含生药）7.14，14.28，28.55 g·kg-1，氯化钠注射液阴性对照组和环磷酰胺阳性对照组，每组10只ICR小鼠，雌雄各半，间隔24 h实施2次灌胃给药，制作骨髓涂片，镜检每只小鼠2 000个嗜多染红细胞中含微核的嗜多染红细胞数，计算每组动物嗜多染红细胞微核率。结果：MLA试验，-S9条件下各浓度组诱发的总突变率呈现浓度依赖性增加，13.65，6.83 g·L-1组与阴性对照组比较有统计学意义 （P〈0.01），小集落突变百分率随浓度增加而升高，＋S9条件下的各浓度组总突变率和小集落突变百分率均与阴性对照组相近；MNT试验，各剂量组无明显骨髓抑制作用，诱发的小鼠骨髓嗜多染红细胞微核率与阴性对照组比较未见明显增加。结论：秦皮水煎剂在体外非代谢活化条件下可诱发L5178Y细胞tk＋/-位点突变并导致染色体损伤，提示其可能存在直接诱变物；秦皮水煎剂对小鼠骨髓细胞染色体无明显损伤作用，经体内、外代谢活化后均未显示遗传毒作用。
Objective： To investigate the genotoxicity of Cortex Fraxini decoction. Method： Mouse lymphoma assay （MLA） and mouse bone marrow micronucleus test （MNT） were used. In MLA, four dose levels of 1.71, 3.42, 6.83 and 13.65 g （crude drug）·L-1 were exposed with L5178Y cells for 3 hours with and without metabolic activation, then expressed for 2 days. The mutation frequency plates were prepared and incubated for 12-13 days. Colony size in each plate was scored, and the total mutation frequency and the percentage of small colony mutants were calculated. In MNT, contained three dose levels of 7.14, 14.28 and 28.55 mg （crude drug）·kg-1 and 10 ICR mice （5 males/5 females） were in each group. The mice were given in every 24 hours by oral gavage twice and sacrificed after 24 hours of the last dosing. Both femur bones were collected to prepare the smear. For each mouse, the number of micronucleated polychromatic erythrocytes （MNPCE） in 2 000 polychromatic erythrocytes was counted, and the mean of rate of MNPCE of each group was calculated. Result： In MLA, the results indicated that the total mutation frequency of four dose levels showed a dose-dependent increase, there was statistically significant difference at high concentrations compared with negative control （P〈0.01）, and the percentage of small colony mutants was similar with positive control in the absence of metabolic activation. The total mutation frequency of each dose level was similar with negative control in the presence of metabolic activation. In MNT, the results indicated that the decoction did not show inhibitory action for bone marrow, and the induced rate of MNPCE of each group was not significantly increased comparing with negative control. Conclusion： Cortex Fraxini decoction induces the tk＋/- gene mutation and chromosome damage in L5178Y cells in vitro without metabolic activation, it hints that the direct mutagens may be within the test article. Cortex Fraxini decoction does not show chromosome damage of bone marr
出处
《中国中药杂志》
CAS
CSCD
北大核心
2009年第17期2228-2231,共4页
China Journal of Chinese Materia Medica
基金
国际科技交流与合作项目(2006DFB31690)
关键词
秦皮水煎剂
小鼠淋巴瘤细胞试验
小鼠骨髓微核试验
遗传毒性
Cortex Fraxini decoction mouse lymphoma assay （MLA） mouse bone marrow micronucleus test （MNT） genotoxicity
