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[目的]选用目前新药遗传毒性评价中推荐使用的3种试验方法(小鼠骨髓微核试验、鼠伤寒沙门氏菌回复突变试验、体外CHL细胞染色体畸变试验)研究盐附子的遗传毒性。[方法]小鼠骨髓微核试验设3个盐附子给药剂量组(15.45、7.73、3.86g生药/kg)、阴性对照组和阳性对照组(环磷酰胺40mg/kg)。Ames试验选用两种方法:直接法和代谢活化法。试验菌株为鼠伤寒沙门氏组氨酸缺陷型菌株TA97、TA98、TA100、TA102、TA1535,盐附子在试验中设6个剂量组(5.000、2.500、1.250、0.625、0.313、0.156mg生药/皿),同时设自发回复突变对照组、阳性对照组。体外CHL细胞染色体畸变试验也选用两种方法:直接法和代谢活化法。盐附子不加S9时处理终浓度为5、2.5、1.25、0.625、0.313mg生药/ml,加S9时处理终浓度为2.5、1.25、0.625、0.313、0.156mg生药/ml。[结果]小鼠骨髓微核试验盐附子各剂量组小鼠骨髓多染红细胞微核率与阴性对照组相比,差异无统计学意义(P﹥0.05)。Ames试验盐附子在加或不加肝微粒体酶(S9)时均未见引起TA97、TA98、TA100、TA102、TA1535试验菌株基因突变,即Ames试验阴性,试验重复一次,所得结论相同。体外CHL细胞染色体畸变试验盐附子在加或不加S9时均未引起CHL细胞的染色体畸变,试验结果为阴性,试验重复一次,所得结果相同。[结论]在实验室条件下,盐附子在小鼠骨髓微核试验、Ames试验及体外CHL细胞染色体畸变试验中均为阴性结果,故认为盐附子无遗传毒性。
[Objective] To investigate the genotoxicity of Aconite Acropoda by using three recommended test methods (micronucleus test of mouse bone marrow, reversion test of Salmonella typhimurium, and chromosome aberration test of CHL cells in vitro) recommended in the current evaluation of genetic toxicity of new drugs. [Methods] Mouse bone marrow micronucleus test was set up with 3 doses of salt aconite administration group (15.45, 7.73, 3.86g crude drug/kg), negative control group and positive control group (cyclophosphamide 40mg/kg). The Ames test uses two methods: direct and metabolic activation. The experimental strains were Salmonella typhimurium-deficient strains TA97, TA98, TA100, TA102, and TA1535. Salt aconite was set in the experiment with 6 dose groups (5.000, 2.500, 1.250, 0.625, 0.313, 0.156 mg crude drug/dish). At the same time, spontaneous reversion mutation control group and positive control group were set up. In vitro CHL chromosome aberration tests also used two methods: direct method and metabolic activation method. Salt aconite without S9 treatment when the final concentration of 5,2.5,1.25,0.625,0.313mg crude drug / ml, plus S9 when the final concentration of 2.5,1.25,0.625,0.313,0.156mg crude drug / ml. [Results] The micronucleus test of mouse bone marrow micronucleus test showed that there was no significant difference in the micronucleus rate of bone marrow multi-erythrocytic murine erythrocytes in each dose group (P>0.05). Ames test salt aconite with or without liver microsomal enzymes (S9) did not cause TA97, TA98, TA100, TA102, TA1535 test gene mutations, Ames test was negative, the test was repeated once, the same conclusions. In vitro Chromosome Aberration Test of CHL Cells Salt aconite had no chromosomal aberrations in CHL cells with or without S9. The test result was negative, and the test was repeated once. The results were the same. [Conclusion] Under laboratory conditions, salt aconite was negative in mouse bone marrow micronucleus test, Ames test, and in vitro CHL cell chromosomal aberration test, so salt aconite was not considered to be genetically toxic.