摘要
目的:研究双氢青蒿素(DHA)对Raji细胞放射敏感性的影响并探讨其作用机制。方法:CCK8测定DHA对Raji细胞活力的影响,流式细胞术检测细胞凋亡、胞内ROS及线粒体膜电位,Western blot检测AKT、pAKT、Bcl-2、Bax和Cleaved-Caspase-3蛋白表达量。结果:实验分为对照组、DHA组(5μmol/L DHA)、放射组(4Gyγ射线)、联合放射组(5μmol/L DHA和4 Gyγ射线),与其他3组相比,联合放射组Raji细胞的线粒体膜电位显著降低(P<0.01),胞内ROS含量和凋亡率显著升高(P<0.01);此外,Raji细胞AKT表达量与其他3组相比无明显差异,但AKT的磷酸化受到抑制;Bcl-2表达量显著降低,而Bax、Cleaved-Caspase-3表达量显著升高。结论:DHA可能通过抑制磷酸肌醇3-激酶(PI3K-AKT)信号通路及激活了Raji细胞的线粒体凋亡途径,引起氧化应激反应,从而增加Raji细胞对放射的敏感性。
Objective: To study the effects of dihydroartemisinin (DHA) on radiation sensitivity of Raji cells, and explore its mechanisms. Methods: CCK8 was used to determine the effect of DHA on cell viability of Raji cells; apoptosis, intracellular reactive oxygen speies (ROS) and mitochondrial membrane potential of Raji cells were detected by flow cytometry; and the protein expressions of protein kinase B (AKT), phospho-rylated-protein kinase B(p-AKT), Bcl-2 and Bax were determined by Western blot. Results: The cells were randomly divided into four groups: control group, DHA(5μmol/L DHA), irradiation(IR, 4 Gy), IR + DHA group (4 Gy IR + 5 μmol/L DHA). Compared with the other three groups, cells in DHA + IR group exhibited lower mitochondrial membrane potential ( P 〈 0.01). While the intracellular ROS content and apoptosis rate of Raji cells in DHA + IR group were increased significantly( P 〈 0.01). In addition, compared with the other three groups, there was no significant difference in the expression of AKT, but the phosphorylation of AKT protein were significantly inhibited and the expression of Bcl-2 protein was markedly decreased. However, the expressions of Bax and Cleaved-Caspase-3 protein were markedly increased. Conclusion: DHA might activate the mitochondrial apoptotic signal via inhibiting phosphoinositide 3-kinase (PI3K/AKT) pathway and increase oxidative stress to enhance the radiosensitivity of Raji cells.
出处
《中国应用生理学杂志》
CAS
CSCD
2017年第5期385-390,共6页
Chinese Journal of Applied Physiology
基金
国家自然科学基金资助项目(81270568)