Yan Xiao-jing, Yang Ye, Bi Lei, Chen Shan-shan, Zhu Jing-jing, Chen Wei-ping
Zhongguo Zhong Yao Za Zhi. 2014 Nov;39(22):4436-41.
This study aims to optimize the most effective component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on lung cancer A549 using the orthogonal design method, and to investigate its effects of the component formula on cell proliferation, apoptosis and cytoskeleton in lung cancer A549 cells. The orthogonal design method was introduced to optimize the most effective component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on lung cancer A549 cells. CCK-8 assay and Real-time cell analysis were adapted to analyze the effect of component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on A549 cells viability at different time and dose. Cell apoptosis was measured by Annexin V- FITC/PI double staining and flow cytometry. Cell skeleton protein F-actin was detected by high content screening (HCS). The optimizing component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma for total salvianolic acid, total saponins of panax ginseng and ginseng polysaccharide doses were 5, 10, 5 mg L(-1). CCK-8 assay and real-time cell analysis demonstrated that the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma treatment could significantly decrease the A549 cell viability in both dose- and time-dependent manner compared with control group (P < 0.01). Moreover, the increase of cell apoptosis was detected by Annexin V-FITC/PI double staining and flow cytometry when cells treated with the component formula, which indicating that the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma could induce A549 cell apoptosis in a time-dependent manner compared with control group (P < 0.01). Furthermore, compared with control group, a significant decrease in A549 cell skeleton area was found in the component formula-exposed cells in the dose-dependent manner (P < 0.01). In summary, the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma inhibits A549 cell proliferation by inducing cell apoptosis and decreasing cell microfilament formation. All of these results will be helpful to reveal antitumor mechanism of the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma, which provides a basis for the exploration of antitumor mechanism of the component formula on lung cancer.
本研究旨在采用正交设计法优化丹参和人参对肺癌A549细胞的最有效成分配方，并研究该成分配方对肺癌A549细胞增殖、凋亡和细胞骨架的影响。引入正交设计法优化丹参和人参对肺癌A549细胞的最有效成分配方。采用CCK-8法和实时细胞分析法分析丹参和人参成分配方在不同时间和剂量下对A549细胞活力的影响。通过Annexin V-FITC/PI双染和流式细胞术检测细胞凋亡。通过高内涵筛选（HCS）检测细胞骨架蛋白F-肌动蛋白。丹参总酚酸、人参总皂苷和人参多糖剂量的丹参和人参优化成分配方分别为5、10、5 mg L(-1)。CCK-8法和实时细胞分析表明，与对照组相比，丹参和人参成分配方处理能以剂量和时间依赖的方式显著降低A549细胞活力（P < 0.01）。此外，用该成分配方处理细胞时，通过Annexin V-FITC/PI双染和流式细胞术检测到细胞凋亡增加，这表明与对照组相比，丹参和人参成分配方能以时间依赖的方式诱导A549细胞凋亡（P < 0.01）。此外，与对照组相比，在成分配方处理的细胞中发现A549细胞骨架面积以剂量依赖的方式显著减少（P < 0.01）。综上所述，丹参和人参成分配方通过诱导细胞凋亡和减少细胞微丝形成来抑制A549细胞增殖。所有这些结果将有助于揭示丹参和人参成分配方的抗肿瘤机制，为探索该成分配方对肺癌的抗肿瘤机制提供依据。
