昆明山海棠总生物碱诱导小鼠淋巴瘤细胞tk基因突变的研究

Effects of Total Alkaloids of Tripterygium Hypoglaucum Hutch on tk Gene of Mouse Lymphoma Cells

目的 :研究昆明山海棠 (THH)总生物碱对小鼠淋巴瘤L5 178Y细胞tk基因的影响。方法 :用抽提的THH总生物碱对L5 178Y细胞进行浓度梯度染毒 ,不同时间点进行单细胞微孔接种 ,计数阳性集落 ,测定细胞接种效率、相对存活率、相对悬浮增长率和突变频率。结果 :随着THH总生物碱作用剂量的增加 ,L5 178Y细胞相对存活率和相对悬浮增长率均明显下降 ;在 0 .1～ 2 .0 g·L-1范围内可诱导细胞tk位点的突变 ,诱发突变频率是自发突变频率的 2～ 9倍。在tk位点诱发了两种不同表型的突变集落 ,即大集落与小集落 ,但以大集落为主 ,这一现象可能与THH的作用机制有关。结论 :THH总生物碱对L5 178Y细胞具有肯定的细胞毒性与诱导基因突变作用。

Objective: To investigate the effects of total alkaloids of tripterygium hypoglaucum hutch (THH) on the tk gene of mouse lymphoma cells. Method: L5178Y cells were infected with total alkaloids of THH with different concentrations and put into single-cell wells at different time phases. Then the numbers of positive wells were counted and the cell plating efficiency, relative suspension growth and mutation frequency were determined. Result: Total alkaloids of THH (0.1～2.0 g·L -1 ) induced tk locus mutation with mutation frequency 2～9 times higher than that of spontaneous mutation frequency of L5178Y cells. There were two different phenotypes of mutation colonies, large colony and small colony, but the main colong was large colony. This phenomenon might be related with the mutagenesis of THH. Conclusion: Total alkaloids of THH can exert toxicity and mutagenic effects on tk gene in L5178Y cells, and there may be range limit in gene mutation.