复方益智散对PC12细胞增殖和抗凋亡作用

Effect of Yi Zhi San on cell proliferation and apoptosis in cultured PC12 cells

目的观察复方益智散对体外培养PC12细胞增殖和抗活性氧(reactive oxygen species,ROS)氧化损伤作用。方法SD大鼠60只,随机分为对照组、复方益智散大、中、小剂量组和复方丹参片组,每组12只。通过血清药理学方法制备含药血清,以过氧化氢(H2O2)作为ROS,应用四氮唑蓝比色法、流式细胞术法和DNA凝胶电泳法观察复方益智散对正常生长PC12细胞的增殖和抗ROS凋亡损伤作用。结果复方益智散中剂量和高剂量组能显著提高正常生长PC12细胞的存活数(P<0.01);各剂量组均能有效对抗100μmol/L H2O2引起的细胞存活率下降和细胞凋亡率升高(P<0.01),减轻DNA片断化现象,复方益智散的抗凋亡效果好于复方丹参片。结论复方益智散具有良好的神经营养和神经保护作用,其机制可能与提高细胞的清除ROS能力和抗氧化酶活力有关。

Objectives To study the effect of Yi Zhi San on cell proliferation and oxidative damage induced by reactive oxygen species （ROS） in cultured PC12 ceils. Methods Sixty SD rats were randomly divided into control group, large, middle and low dosage groups of Yi Zhi San, and compound Salvia Miltiorrhiza tablet group. Each group included 12 rats. The sera containing Yi Zhi San were obtained from rats by using the serum pharmacology method. Hydrogen peroxide was used as ROS and MTT, flow cytometry and DNA gel electrophoresis methods were used to evaluate the cell proliferation and apoptosis of the cultured PC12 cells. Results Yi Zhi San in middle and large dose groups could increase PC12 cell proliferation, but the low dose group could not. All of the three dosages could resist the decrease in the cell viability and increase in the apoptotic rate and inhibit the DNA fragmentation induced by 100 μmol/L H2O2 treatment （ P 〈 0.01）. These effects were superior to those achieved by compound Salvia Miltiorrhiza tablet （ P 〈 0.01）. Conclusion Yi Zhi San has the great potential of neurotrophic and neuroprotective effects and the mechanisms may be related to enhancing the abilities of ceils to scavenge ROS and to resist the oxidative damage.