灯盏乙素对PC12细胞拟缺血性损伤的保护作用研究

Protective Effect of Scutellarin on Ischemic Injury in PC12 Cells

目的:研究灯盏乙素对PC12细胞拟缺血性损伤的保护作用,并探讨其作用机制。方法:用连二亚硫酸钠(Na2S2O4)加缺糖复制PC12细胞拟缺血模型。以MTT法检测细胞存活率并进行乳酸脱氢酶(LDH)活力测定;应用Hoechst33342染色法证明凋亡细胞的存在;应用流式细胞仪检测凋亡细胞的比率及线粒体膜电位的变化;应用激光共聚焦显微镜观察灯盏乙素作用下细胞内钙离子(Ca2+)浓度的变化。结果:在10-7～10-5mol.L-1范围内,灯盏乙素可增加损伤细胞的存活率,降低缺血性损伤所致培养介质内LDH的释放,具有浓度依赖性;灯盏乙素还可降低PC12拟缺血性损伤细胞的凋亡百分率和稳定细胞线粒体跨膜电位,降低细胞内Ca2+浓度,具有浓度依赖性。结论:灯盏乙素对PC12细胞拟缺血性损伤有显著保护作用,其机制可能与稳定细胞线粒体跨膜电位与抑制Ca2+内流有关。

OBJECTIVE： To investigate the protective effect of scutellarin on ischemic injury in PC12 cells and explore its action mechanisms. METHODS： PC12 cells were treated with Na2SO4 in combination with sugar deficiency to induce ischemic injury model. The survival rate of the PC12 cells were detected using MTT assay; and the activity of lactate dehydrogenase （LDH） were detected. The apoptosis was confirmed by Hoechst 33342 staining and flow cytometric assay was performed to determine the apoptotic rate and the mitochondrial membrane potential（MMP） in PC12 cells. The concentration of cellular calcium ion （Ca^2 ＋） was detected by laser scanning confocal microscopy. RESULTS： Within the range of 10^- 7 10 ^-5 mol · L^- 1, Scutellarin significantly increased the survival rate of the injured cells, decreased ischemie injury - induced LDH release within the cultured medium in a concentration dependent manner, decreased the apoptotic rate, stabilized mitochondrial transmembrane potential and decreased the intracellular concentration of Ca^2＋ in concentration- dependent manner. CONCLUSION： Scutellarin has remarkable protective effect on PC12 cells against the ischemic injury. The mechanism of the protection might be attributed to its action of stabilizing mitochondrial transmembrane potential and inhibiting on the inflow of Ca^2＋.