五味子水煎剂的遗传毒性研究

Genotoxicity of Fructus Schisandrae Chinensis Concoction

背景与目的:采用小鼠淋巴瘤细胞试验(MLA)和小鼠骨髓微核试验(MNT)研究五味子水煎剂的遗传毒性。材料与方法:在MLA中,设五味子0.97、1.94、3.88、7.77 mg/ml(生药)等4个浓度组,在非代谢活化(-S9)和代谢活化(+S9)条件下处理L5178Y细胞3 h,表达2 d,制备基因突变频率平板并培养12 d,计数含大、小突变细胞集落的孔数,计算基因突变频率(MF)和小集落突变百分率(SC%);在MNT中,设3.13、6.25、12.50 g/kg(生药)3个剂量组,每组10只ICR小鼠,雌雄各半,间隔24 h实施2次小鼠灌胃给予五味子水煎剂,制作骨髓涂片,计数每只小鼠2 000个嗜多染红细胞中含微核的嗜多染红细胞数(MNPCE),计算每组动物嗜多染红细胞微核率。结果:在-S9条件下,7.77 mg/ml(生药)浓度组诱发的MF是阴性对照组的2倍以上(P<0.01),SC%随五味子浓度增加而升高。+S9条件下的各浓度组诱发的MF存在剂量-反应关系,7.77 mg/ml(生药)浓度组的MF明显增加(P<0.01),其MF、SC%与阳性对照组相近;五味子水煎剂各剂量组未显示骨髓抑制作用,所诱发的微核率与阴性对照组比较未见明显增加(P>0.05)。结论:五味子水煎剂在+/-S9条件下均可诱发L5178Y细胞tk位点突变并导致染色体损伤,提示对人体具有潜在的遗传毒性;但对小鼠骨髓细胞染色体无损伤,经体内代谢活化后未显示遗传毒作用。

BACKGROUND AND AIM： To investigate the genotoxicity of Fructus Schisandrae Chinensis concoction using mouse lymphoma assay（MLA） and mouse bone marrow micronucleus test （MNT） . MATERIALS AND METHODS： In MLA, four doses 0.97, 1.94, 3.88, 7.77 mg/ml （crude drug） were used to treat 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 days. Colony size in each plate was scored, and total mutation frequency and percentage of small colony mutants were calculated and assayed. In MNT, three dose levels of 3.13,6.25,12.50 g/kg （crude drug）were used.10 ICR mice （5 males/ 5 females） in each group were gavaged every 24 h twice and sacrificed after 24 hours of the last dosing. Both femur bones were collected to prepare bone marrow smear. For each mouse, the number of micronucleated polychromatic erythrocytes （MNPCE） in 2000 polyehromatic erythrocytes （PCE） was counted, and the mean of rate of MNPCE of each group was calculated. RESULTS： MLA indicated that the total mutation frequency showed a dose-dependent increase with metabolic activation, and was statistically significant （P〈0.01） compared with negative control with and without metabolic activation. Percentage of small colony mutants was similar with positive control at high concentrations. MNT indicated the concoction did not show inhibitory effects on hone marrow, and the rate of MNPCE of each group induced did not show significant increase compared with negative control. CONCLUSION： Fructus Schisandrae Chinensis concoction induced tk gene mutation and chromosomal damage in indicated that the test article may have potential genotoxicity concoction did not show chromosomal damage of bone marrow in LS178Y cells with and without metabolic activation. It for humans. However Fructus Schisandrae Chinensis ICR mice, demonstrating no genotoxicity in vivo with metabolic activation under this study condition.