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目的:确证丹参酮 IIA 的细胞毒依赖于 NAD(P)H 醌氧化还原酶 (NQO1)这一假设。方法:首先进行 MTT试验考察丹参酮 IIA 在一系列具有不同 NQO1 酶活的细胞株的细胞毒, 并考察 NQO1 的特异性抑制剂双香豆素对丹参酮 IIA 细胞毒的逆转来确证丹参酮 IIA 的细胞毒是依赖于 NQO1 的。然后在具有较强丹参酮 IIA 细胞毒的 HepG2 细胞中考察了丹参酮 IIA的生物转化情况, 并考察了双香豆素对丹参酮 IIA 生物转化情况影响。结果:丹参酮 IIA 的细胞毒与细胞株的 NQO1 酶活呈正相关。双香豆素能够逆转丹参酮 IIA 的细胞毒作用, 同时也能抑制丹参酮 IIA 在 HepG2 细胞毒的生物转化。结论:NQO1 是丹参酮 IIA 发挥抗肿瘤效应的首要靶点, 通过抑制 NQO1 双香豆素能够减少丹参酮 IIA 向儿茶酚中间体的转化, 从而打破丹参酮 II在 NQO1 介导下发生还原-自氧化-还原循环, 减少活性氧自由基的产生, 而减少丹参酮 IIA 的细胞毒作用。由此证明, 丹参酮 IIA的细胞毒是由 NQO1-介导其生物转化过程而产生的。
Objective: To confirm the hypothesis that the cytotoxicity of tanshinone IIA is dependent on NAD (P) H quinone oxidoreductase (NQO1). Methods: MTT assay was performed to investigate the cytotoxicity of tanshinone IIA in a series of cell lines with different NQO1 activities. The cytotoxicity of tanshinone IIA was confirmed by reversing the cytotoxicity of tanshinone IIA, a specific inhibitor of NQO1, It depends on NQO1. Then the biotransformation of tanshinone IIA was investigated in HepG2 cells with strong Tanshinone IIA cytotoxicity and the effects of dicoumarin on the biotransformation of tanshinone IIA were investigated. Results: Tanshinone IIA cytotoxicity and cell line NQO1 activity was positively correlated. Dicoumarin reverses the cytotoxic effect of tanshinone IIA and at the same time inhibits the biotransformation of tanshinone IIA in HepG2 cells. CONCLUSION: NQO1 is the primary target of tanshinone IIA to exert anti-tumor effect. The inhibition of NQO1-dicoumarol can reduce the conversion of tanshinone IIA to catechol intermediates, thus breaking down the tanshinone II-mediated reduction-autoxidation- Reduce the cycle, reduce the generation of reactive oxygen free radicals, and reduce the tanshinone IIA cytotoxicity. Thus, it was demonstrated that the cytotoxicity of tanshinone IIA is caused by NQO1-mediated biotransformation.