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目的:探讨丹参素(DSS)抗肿瘤作用的分子机制,重点在于是否与其抗氧化作用及改变非小细胞肺癌A549细胞内氧化还原状态及影响相关核转录因子和信号通路有关。方法:通过体外DPPH自由基清除、高铁还原、亚铁螯合实验和DCFH-DA荧光探针标记ROS,检测DSS的抗氧化活性及对A549细胞内ROS水平的影响;采用MTT法测定DSS对A549细胞增殖能力的时效与量效关系;采用Western blot技术检测DSS对A549细胞内缺氧诱导因子HIF-1α及氧化还原调节转录因子Nrf2蛋白表达的影响,以及上游Akt,ERK1/2 MAPK信号磷酸化水平的影响。结果:DSS能够剂量依赖性降低A549细胞内ROS水平,与其较强的自由基清除和抗氧化活性有关;DSS能以时间和剂量依赖性的方式抑制A549细胞生长,进一步机制研究发现,DSS能降低Nrf2表达,与抑制Akt以及ERK1/2的磷酸化水平有关,但对HIF-1α的表达没有影响。结论:DSS具有治疗非小细胞肺癌的作用,机制在于通过清除ROS进而抑制Akt,ERK1/2的磷酸化而下调Nrf2的表达并最终抑制A549细胞的生长。
AIM: To investigate the molecular mechanism of anti-tumor effects of danshensu (DSS), and to determine whether it is related to its anti-oxidative effects and to the changes of intracellular redox status in A549 cells and the related nuclear transcription factors and signaling pathways. Methods: The anti-oxidative activity of DSS and the effect of DSS on the level of ROS in A549 cells were detected by DPPH radical scavenging, high iron reduction, ferrous chelation and DCFH-DA fluorescent probe in vitro. The effect of DSS on A549 The effect of DSS on hypoxia-inducible factor HIF-1αand redox-regulated transcription factor Nrf2 protein expression in A549 cells was detected by Western blot and phosphorylation of Akt and ERK1 / 2 MAPK signals Horizontal impact. Results: DSS could reduce the level of ROS in A549 cells in a dose-dependent manner, which was related to the strong activity of free radical scavenging and antioxidation. DSS could inhibit the growth of A549 cells in a time and dose-dependent manner. Further study found that DSS could reduce Nrf2 expression, and inhibition of Akt and ERK1 / 2 phosphorylation level, but had no effect on the expression of HIF-1α. Conclusion: DSS can treat non-small cell lung cancer by down-regulating the phosphorylation of Akt and ERK1 / 2 and decreasing the expression of Nrf2 and eventually inhibiting the growth of A549 cells.