摘要
目的探查三黄抗氧化方(SHD)基于Nrf-2/ARE通路的抗氧化反应与肿瘤血管新生的关系。方法将实验分为空白组:MCF-7细胞培养,实验组按黄芪、制大黄、片姜黄(3∶1∶1)不同浓度,分为5组:0.5mg/ml,1.0mg/ml,2.0mg/ml,4.0mg/ml,8.0mg/ml。CCK-8法检测不同浓度三黄抗氧化方对MCF-7细胞增殖率的影响;免疫印迹法(Western Blot)检测MCF-7细胞Nrf-2蛋白的表达;用RNA干扰技术(RNAi)静默Nrf-2基因,Western Blot法检测三黄抗氧化方对siRNA静默MCF-7细胞Nrf-2的影响;酶联免疫吸附实验(ELISA法)检测三黄抗氧化方对Nrf-2siRNAMCF-7细胞VEGF含量水平的影响。结果三黄抗氧化方能够抑制MCF-7细胞增殖;三黄抗氧化方可以上调MCF-7细胞中Nrf-2的表达;三黄抗氧化方可以促进Nrf-2核内转位;三黄抗氧化方可以上调Nfr-2siRNA静默MCF-7细胞中Nrf-2基因表达;三黄抗氧化方可以抑制Nfr-2siRNA静默MCF-7细胞的VEGF水平。结论三黄抗氧化方通过促进Nrf-2蛋白核内转位从而激活抗氧化反应,发挥抑制MCF-7细胞血管新生的作用。
Objective To investigate the relationship between antioxidant response based on Nrf - 2/ARE pathway and tumor angiogenesis of Sanhuang antioxidant decoction （SHD）. Methods The experiment was divided into blank group： MCF - 7 cells were cultured, the experimental group： astragalus root, rhubarb, turmeric （3： 1：1 ） divided into 5 groups of O. 5mg/mL, 1. 0mg/mL and 2.0mg/mL, 40mg/mL, 8.0mg/mL. CCK - 8 method detected MCF -7 cell proliferation rate of SHD. Western Blot was used to detect the expression of Nrf - 2 protein of MCF - 7 cell in the cytoplasm and nuclear. The Nrf - 2 gene was knocked down by RNA interference （RNAi）, and the MCF -7 Nrf- 2siRNA cell of VEGF was detected by enzyme -linked immunosor- bent assay （ELISA）. Results SHD can inhibit the proliferation of MCF- 7 cells SHD can upregulate the expression of Nrf-2 in MCF - 7 cells and transfer to nuclear, SHD can inhibit the expression of VEGF level in MCF - 7 Nrf - 2siRNA cells. Conclusion Antioxidant by promoting Nrf - 2 protein SHD nuclear translocation, therefore inhibit the role of angiogenesis in MCF - 7 cells.
出处
《时珍国医国药》
CAS
CSCD
北大核心
2016年第11期2579-2581,共3页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(No.81503583)
江苏省中医院院级课题
(No.Y140130)
