丹参素对非小细胞肺癌A549细胞内氧化还原状态及相关核转录因子的影响  

Effect of Danshensu on redox state and relevant nuclear transcription factors in non-small cell lung cancer A549 cells

作　　者：陶丽[1] 王生[1] 赵杨[1] 陈文星[1,2] 王爱云[1,2] 陆茵[1,2] 

机构地区：[1]南京中医药大学药学院,江苏南京210046 [2]江苏省中药药效与安全性评价重点实验室,江苏南京210029

出　　处：《中国中药杂志》2012年第9期1265-1268,共4页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81173174;30772766);江苏省自然科学基金项目(BK2010085;210562);江苏省国际合作项目(SBZ200900175);江苏省教育厅重大专项(09KJA360002);江苏省六大人才高峰课题A类第五批项目(08-A-012);江苏省中医药局项目(LZ09022);江苏省中药药效与安全性评价重点实验室开放课题(P09013);江苏省普通高校研究生科研创新计划项目(2010-469;2010-471)

摘　　要：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。Objective： To study the molecular mechanism of anti-tumor effect of Danshensu（DSS） focusing on whether it is related to its antioxidation effect on redox state of non-small cell lung cancer A549 cells and relevant nuclear transcription factors and signal pathway.Method： DPPH radical scavenging,ferric reducing,ferrous iron chelating assay and DCFH-DA fluorescence staining of ROS were introduced to detect antioxidant activity and ROS scavenging activity in vitro;MTT was adopted to analyze the effect of DSS on A549 cells viability at different time and dose.Western blot was used to detect the effect of DSS on the protein expression of hypoxiainducible factor HIF-1α and redox regulating transcription factor Nrf2 in A549 cells and the phosphorylation of upstream Akt,ERK1/2 MAPK.Result： DSS could reduce the ROS level in A549 cells in a dose-dependent manner,which was related to it′s strong DPPH radical scavenging activity.Moreover,DSS inhibited A549 cells proliferation in both dose-and time-dependent manner.Further study on molecular mechanism indicated that DSS suppressed redox regulator Nrf2,and down-regulated the phosphorylation of Akt and ERK1/2.However,DSS had no effect on HIF-1α expression.Conclusion： DSS might have the effect on treating non-small cell lung cancer by scavenging ROS and then inhibiting phosphorylation of Akt,ERK1/2 and down-regulating Nrf2 expression in A549 cells.

关 键 词：丹参素 非小细胞肺癌A549细胞株 ROS NRF2 HIF-1Α 

分 类 号：R285.5[医药卫生—中药学] R734.2[医药卫生—中医学]