杜仲水煎剂的遗传毒性研究  

Genotoxicity study of Ortex Eucommiae decoction

作　　者：胡燕平[1] 王欣[1] 宋捷[1] 张旻[1] 王秀文[1] 李波[1] 

机构地区：[1]中国药品生物制品检定所国家药物安全评价监测中心,北京100176

出　　处：《华西药学杂志》2009年第5期490-493,共4页West China Journal of Pharmaceutical Sciences

基　　金：国际科技合作项目:促进中药材在法国的注册和进入(No.2006DFB31690)

摘　　要：目的研究杜仲水煎剂的遗传毒性。方法采用小鼠淋巴瘤细胞试验(MLA)和小鼠骨髓微核试验(MNT)。在MLA中,2.5、5、10、20 mg.ml-1生药4个浓度组在非代谢活化(-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 d,计数大、小突变细胞集落的孔数,计算总突变率(MF)和小集落突变百分率(SC%);在MNT中,12.8、25.6、51.2 g.kg-1生药3个剂量组间隔24 h灌胃给药2次,制作骨髓涂片,计数每只小鼠两千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的平均微核率。结果4个浓度组在+/-S9条件下诱发的MF呈现剂量相关性增加,与阴性对照组比较有统计学意义(P<0.05),-S9条件下的SC%与阳性对照组相仿,+S9条件下的SC%随浓度增加而升高。各剂量组未显示骨髓抑制作用,诱发的微核率与阴性对照组比较未见明显增加。结论杜仲水煎剂在+/-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;但供试品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。OBJECTIVE To investigate genotoxicity of Ortex eucommiae decoction using mouse lymphoma assay （MLA） and mouse bone marrow micronucleus test （MNT）. METHODS In MLA, four dose levels of 2.5,5,10,20 mg （crude drug） ·mL^-1 was 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 days. Colony size in each plate was scored, and the total mutation frequency and the percentage of small colony mutants were calculated and analyzed. In MNT, different doses （ 12.8,25.6,51.2 g·kg^- 1 ） were administered to three groups. The mice were adminstrated every 24 hours by oral gavage twice and sacrificed after 24 hours of the last administration and both femur bones were collected to prepare the smear. For each mouse, the number of micronucleated polychromatic erythrocytes （MNPCE） in 2000 polychromatic erythrocytes was counted, and the mean rate of MNPCE of each group was calculated. RESULTS In MLA, the results indicated that the total mutation frequency of dose levels showed a dose - dependent increase and there was statistical significant （ P 〈 0. 05 ） compared with negative control with or without metabolic activation ; the percentage of small colony mutants was similar with positive control without metabolic activation, and there was a dose - dependent increase with metabolic activation. In MNT, the results indicated the decoction did not show inhibitory effect for bone marrow, and the rate of MNPCE of each group induced did not show significantly increase compared with negative control. CONCLUSION Ortex Eucommiae decoction induced tk gene mutation and chromosome damage in LS178Y cells with and without metabolic activation,suggesting that the test article may have potential genotoxicity for human bodies. Ortex Eucommiae decoction did not show chromosome damage of bone marrow cells in ICR mice, and it had not genotoxicity in vivo with metabolic activation in our experiment.

关 键 词：杜仲水煎剂 淋巴瘤细胞试验 骨髓微核试验 遗传毒性 

分 类 号：R96[医药卫生—药理学]