利用中国仓鼠肺细胞体外微核试验评价三七提取液的遗传毒性

In vitro induction of micronuclei by Panax notoginseng extracting solution

目的:评价传统中药三七提取液的遗传毒性.方法:采用体外胞质分裂阻断微核法检测三七提取液在加与不加代谢活化系统S9的两种条件下对中国仓鼠肺细胞微核率的影响,三七提取液的制备采用乙醇反复浸提法.根据中性红摄取法细胞毒性测定试验的结果,微核试验设3个三七染毒浓度组(0.625、1.250、2.500 mg/mL)、阳性对照组(+S9条件下为20μg/mL环磷酰胺,-S9条件下为1μg/mL丝裂霉素C)和阴性对照组(MEM培养基).结果:三七提取液2.500 mg/mL浓度组,中性红摄取法细胞毒性测定试验中,吸光度值较阴性对照组显著降低(P<0.05),细胞存活率为80.3%;三七提取液0.625、1.250、2.500 mg/mL浓度组CHL细胞的微核率,在+S9条件下分别为17‰、16‰、16‰,在-S9条件下分别为17‰、15‰、16‰.与阴性对照组(+S916‰、-S915‰)比较,差异均无统计学意义(P>0.05).结论:在本研究条件下,体外微核试验结果表明,三七提取液不具有遗传毒性.

OBJECTIVE: The in vitro micronucleus test was applied to evaluate genotoxicity of the traditional Chinese medicine Panax notoginseng extracting solution. METHODS: The Chinese hamster lung cell line (CHL) were treated with Panax notoginseng extracting solution which was prepared by soaking with alcohol repeatedly at three concentrations (0.625, 1.250, 2.500 mg/mL). Cell toxicity was determined by the neutral red uptake test. Genotoxicity was determined with and without the S9 metabolic activation via the in vitro cytokinesis-block micronucleus test. Cyclophosphamide and mitomycin C were used as the positive controls,and MEM was used as the negative control. RESULTS:The neutral red uptake test showed that the 2.500 mg/mL concentration of Panax notoginseng extracting solution induced OD value decreased significantly compare with negative control (P<0.05) which indicates that the cell survival rate was 80.3%. The frequencies of micronuclei in the three concentrations of 0.625, 1.250, 2.500 mg/mL of Panax notoginseng extracting solution were 17‰,16‰,16‰ in the presence of +S9. In the absence of S9,the frequencies were 17‰, 15‰, 16‰ which were not significantly different from the negative control (+ S9 16‰、- S9 15‰, P>0.05). CONCLUSION:Panax notoginseng extracting solution did not show genotoxicity under our study conditions.