Hu Yanping, Song Jie, Wang Xin, Zhang Min, Wang Xiuwen, Li Bo
National Center for Safety Evaluation of Drugs, National Institute for the Control of Pharmaceutical and Biological Products, Beijing, 100176, China.
Zhongguo Zhong Yao Za Zhi. 2009 Sep;34(17):2228-31.
To investigate the genotoxicity of Cortex Fraxini decoction.
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) x L(-1) were exposed with I5178Y 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) x 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.
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.
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 marrow in ICR mice, it has not genotoxicity in vitro/in vivo with metabolic activation under this study condition.
研究秦皮水煎液的遗传毒性。
采用小鼠淋巴瘤试验(MLA)和小鼠骨髓微核试验(MNT)。在MLA中,将1.71、3.42、6.83和13.65 g(生药)×L⁻¹这四个剂量水平与I5178Y细胞在有和无代谢活化的情况下接触3小时,然后培养2天。制备突变频率平板并孵育12 - 13天。对每个平板中的集落大小进行评分,并计算总突变频率和小集落突变体的百分比。在MNT中,设置7.14、14.28和28.55 mg(生药)×kg⁻¹这三个剂量水平,每组10只ICR小鼠(5只雄性/5只雌性)。小鼠每24小时经口灌胃给药两次,末次给药24小时后处死。收集双侧股骨制备涂片。对每只小鼠,计数2000个多染红细胞中的微核多染红细胞(MNPCE)数量,并计算每组MNPCE率的平均值。
在MLA中,结果表明四个剂量水平的总突变频率呈剂量依赖性增加,高浓度时与阴性对照相比有统计学显著差异(P < 0.01),在无代谢活化时小集落突变体的百分比与阳性对照相似。在有代谢活化时,各剂量水平的总突变频率与阴性对照相似。在MNT中,结果表明该水煎液对骨髓无抑制作用,每组MNPCE的诱导率与阴性对照相比无显著增加。
秦皮水煎液在无代谢活化的情况下可诱导L5178Y细胞中的tk(+/-)基因突变和染色体损伤,提示受试物中可能存在直接诱变剂。秦皮水煎液在ICR小鼠中未显示骨髓染色体损伤,在本研究条件下其在有代谢活化时体外/体内均无遗传毒性。