氧化应激诱导髓核细胞凋亡中细胞内活性氧及线粒体膜电位的变化

Changes of ROS and mitochondrial transmembrane potential in oxidative stress-induced apoptosis in nucleus pulposus cells of rats

目的:研究氧化应激诱导的髓核细胞凋亡中细胞内活性氧(ROS)及线粒体膜电位的变化。方法:原代培养大鼠髓核细胞,以不同浓度过氧化氢(H2O2)作用于髓核细胞制造氧化应激模型,CCK8法检测H2O2诱导髓核细胞损伤过程中的细胞存活率,Annexin V/PI流式细胞术检测细胞凋亡率,DCFH-DA荧光定量法检测ROS含量、JC-1染色检测线粒体膜电位。结果:CCK8法检测H2O2能够降低髓核细胞生存率并呈剂量相关性,流式细胞术检测发现随着H2O2浓度的升高细胞凋亡比例增多,细胞内ROS增加,线粒体膜电位下降。结论:ROS介导的线粒体膜电位下降参与氧化应激诱导的髓核细胞凋亡,ROS有望成为延缓椎间盘退变的治疗靶点。

Objective To investigate the changes of ROS and mitochondrial transmembrane potential （MMP） in oxidative stress-induced apoptosis in nucleus pulposus cells （NPC） of rats. Methods Nucleus pulposus cells of rats were cultured and then were administered with different concentrations of hydrogen dioxide （H2O2） for 6 h. Then the survival rate of the cells was examined by CCK8 ; the apoptotic rate was defined by Annexin V/PI flow cytometric analysis; intraeellular level of ROS was measured by DCFH-DA fluorescent quantitation; and MMP was detected by JC- 1 staining. Results With the increase of the concentration of H2O2, the survival rate of NPC was decreased, the apoptotic rate was increased, intracellular level of ROS was increased, and the MMP was decreased. Conclusion ROS-mediated MMP loss played an important role in oxidative stress-induced apoptosis in NPC, and may serve as a potential target in the treatment for disc degeneration.