吴茱萸及其主要成分的遗传毒性研究

摘要

目的：研究吴茱萸及其主要成分吴茱萸碱、吴茱萸次碱和柠檬苦素的遗传毒性，为有毒中药吴茱萸的开发利用提供依据。方法：选用了Ames试验、体外CHL细胞染色体畸变试验和小鼠骨髓嗜多染红细胞微核试验。 Ames 试验选用TA97、TA98、TA100、TA102、TA1535组氨酸缺陷型菌株。吴茱萸碱、吴茱萸次碱和柠檬苦素分别设5个剂量组：0.0005、0.005、0.05、0.5、5 mg/皿；体外CHL细胞染色体畸变试验，吴茱萸碱、吴茱萸次碱和柠檬苦素的剂量分别为0.005 mg/mL、0.05/mL和0.5 mg/mL，受试药物与CHL细胞接触时间分别为4 h和24 h；吴茱萸醇提物小鼠骨髓微核试验，设吴茱萸醇提物0.88、3.52、10.55 g （生药）/kg 3个给药剂量组。连续给药4 d，每天给药1次。结果：吴茱萸碱、吴茱萸次碱和柠檬苦素各剂量组Ames试验结果为阴性。 CHL试验中，吴茱萸碱的试验结果为阴性；吴茱萸次碱24 h组细胞染色体畸变率略有增加，差异有统计学意义（ P＜0.05）；柠檬苦素4 h和24 h组细胞染色体畸变率都略有增加，0.05 mg/mL、0.5 mg/mL组与阴性对照组比较，差异有统计学意义（P＜0.05），而0.005 mg/mL组与阴性对照比较，差异无统计学意义（P＞0.05）。微核试验中，吴茱萸醇提物的小鼠骨髓细胞微核试验结果为阴性。结论：综合以上3个试验结果，认为在本实验室条件下，吴茱萸醇提物无遗传毒性。但在体外试验中，吴茱萸次碱和柠檬苦素有致突变性。为进一步确认吴茱萸及其主要成分的遗传毒性，可进一步研究和探讨。%Objective:To investigate the genetic toxicity of evodia rutaecarpa and its main components , including evodiamine , rutae-carpin and limonin , in order to provide useful data for developing toxic Traditional Chinese Medicine .Methods:Ames test, CHL chro-mosome aberation assay and micronucleus assay of bone marrow cell in mice were used to test evodiamine , rutaecarpin , limonin and evo-dia rutaecarpa.Strains TA97,TA98,TA100,TA102 and TA1535 were used in the Ames test.There were five dose groups respectively for evodiamine, rutaecarpin and limonin as follows:0.0005, 0.005 mg/plate, 0.05 mg/plate, 0.5 mg/plate, 5 mg/plate.As a result, the results of the Ames assay were negative for various groups of evodiamine , rutaecarpin and limonin .In CHL chromosome aberration assay, the doses of evodiamine , rutaecarpin and limonin were 0.005 mg/mL, 0.05 mg/mL and 0.5 mg/mL respectively.Exposure time were 4 h and 24 h.Results:The result of the Evodiamine was negative .Aberration rate of 24 h exposure Rutaecarpin chromosome in-creased significantly compared with that of negative control group (P<0.05).Aberration rate of Limonin chromosome 0.05 and 0.5 mg/ml groups increased in 4 h and 24 h exposure compared with negative control group (P<0.05), the 0.005 mg/mL group was not significantly different compared with negative control group (P>0.05).In vivo micronucleus test with evodia rutaecarpa alcohol extract , there were 3 dosing groups:0.88, 3.52,10.55 g raw drug/kg, 4 days, once every day.The results was negative.Conclusion:no genet-ic toxicity was observed in evodia rutaecarpa alcohol extract .However , rutaecarpin and limonin shows some genotoxicity in vivo test , which requires further research to confirm .