黄芩苷和黄芩素通过 ERK/p38 MAPK 通路增强人乳腺癌细胞凋亡。
The combination of baicalin and baicalein enhances apoptosis via the ERK/p38 MAPK pathway in human breast cancer cells.
机构信息
Research Center for Traditional Chinese Medicine Complexity System, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
出版信息
Acta Pharmacol Sin. 2009 Dec;30(12):1648-58. doi: 10.1038/aps.2009.166.
AIM
To examine whether the cell growth inhibitory effect of the combination of baicalin and baicalein is related to apoptosis. Moreover, to determine whether the expression of some apoptosis-related proteins is regulated by the ERK/p38 MAPK pathway.
METHODS
Cell viability was measured using a 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was detected by acridine orange (AO) staining, DNA ladder assay and flow cytometric analysis. Apoptosis-related proteins were observed using Western blot analysis.
RESULTS
Compared with baicalin or baicalein alone, the combination treatment of baicalin (50 micromol/L) and baicalein (25 micromol/L) had an anti-proliferative effect in a time-dependent manner. Isobologram analysis demonstrated that the combination treatment had a synergistic effect. Moreover, apoptosis in MCF-7 cells was increased by 12% and 20% with the combination treatment at 24 h and 48 h, respectively. With the combination treatment in MCF-7 cells, cleaved caspase-3 and caspase-9 were observed, and the level of bcl-2 expression was decreased approximately 20% and 40% at 24 h and 48 h, respectively. The expression of bax and p53 were increased about 25% and 15% at 48 h, respectively. Moreover, the activation of caspase-3, -9 and the regulation of bcl-2, bax and p53 were related to ERK /p38 MAPK activation.
CONCLUSION
In this study, apoptosis was enhanced by the combination treatment of baicalin and baicalein, which activated caspases-3 and caspase-9, downregulated the level of bcl-2 and upregulated the level of bax or p53 via the ERK/p38 MAPK pathway.
目的
研究黄芩苷和黄芩素联合应用的细胞生长抑制作用是否与细胞凋亡有关，同时探讨某些细胞凋亡相关蛋白的表达是否受细胞外信号调节激酶（ERK）/p38 丝裂原活化蛋白激酶（p38 MAPK）通路调控。
方法
采用 3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）比色法检测细胞活力，吖啶橙（AO）染色、DNA 梯状电泳和流式细胞术检测细胞凋亡，Western blot 检测凋亡相关蛋白。
结果
与黄芩苷或黄芩素单独处理相比，黄芩苷（50 μmol/L）和黄芩素（25 μmol/L）联合处理呈时间依赖性抑制 MCF-7 细胞增殖，棋盘格分析表明两药联合具有协同作用。联合用药 24 h 和 48 h 时 MCF-7 细胞凋亡率分别增加了 12%和 20%。联合用药可激活 caspase-3 和 caspase-9，24 h 和 48 h 时 bcl-2 蛋白表达分别下降约 20%和 40%，bax 和 p53 蛋白表达分别增加约 25%和 15%。此外，caspase-3、caspase-9 的激活及 bcl-2、bax 和 p53 的调控与 ERK/p38 MAPK 的激活有关。
结论
本研究表明黄芩苷和黄芩素联合应用可通过激活 ERK/p38 MAPK 通路，增强 caspase-3、caspase-9 的活性，下调 bcl-2 蛋白水平，上调 bax 或 p53 蛋白水平，从而促进 MCF-7 细胞凋亡。
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