【摘要】 目的以原代培养的胚胎大鼠大脑皮质神经细胞进行缺氧缺糖再给氧处理,观察党参皂苷L1对神经细胞凋亡的抑制作用。方法胚胎大鼠大脑皮质神经细胞原代培养,含连二亚硫酸钠的无糖Earle s液进行缺氧缺糖处理造模,MTT法测定细胞存活率,Hoechst33342和PI原位双染法荧光显微镜观察细胞形态学变化和坏死、凋亡细胞百分率,APO BRDU法流式细胞术检测凋亡细胞百分率,Fluo3法流式细胞术检测细胞内Ca2+浓度,评价药效。结果(1)细胞存活率(%)与模型组(29.89±6.28)比较党参皂苷L11.0mg L组(46.05±8.92)、0.1mg L组(46.83±6.76)、0.01mg L组(41.05±9.18)均能显著提高细胞存活率(P<0.01和P<0.05);(2)细胞坏死率(%)与模型组(44.99±12.81)比较,党参皂苷L11.0mg L组(19.45±2.31)、0.1mg L组(18.78±4.95)和0.01mg L组(22.37±4.44)能显著降低细胞坏死率(P<0.001);(3)原位双染法细胞凋亡率(%)与模型组(17.44±4.82)比较,党参皂苷L11.0mg L组(8.75±1.88)、0.1mg L组(9.73±1.76)能显著降低细胞凋亡率(P<0.01和P<0.05);(4)流式细胞术检测细胞凋亡率(%)与模型组(8.55±3.84)比较,党参皂苷L11.0mg L组(4.21±2.31)、0.1mg L组(3.83±2.95)能显著降低细胞凋亡率(P<0.05);(5)细胞内Ca2+平均荧光值,与模型组(47.37±9.68)比较,?更多还原

【Abstract】 Objective:To study the protective effect and its mechanism of saponin L1 of Radix Codonopsis(RCS-L1) on the neuronal death and apoptosis induced by hypoxia/hypoglycemia and reoxygenation in cultured rat primary cell culture nerve cells,and to supply experimental foundation of the therapeutic princirple of“supporting health energy,strengthening body resistance and protecting brain” for the treatment in the acute stage of apoplexic stroke.Method:The model of hypoxia/hypoglycemia and reoxygenation induced damage was set up in vitro by sodium dithionate.The cell survival rate was detected by MTT assay.Morphological changes of neuronal necrosis and apoptosis were observed with fluorescence microscope.Apoptosis and intracellular free calcium ion concentration([Ca~2+ ]i) were measured by flow cytometry.Results:Hypoxia/hypoglycemia cultures for 1 hour and reoxygenation induced neuronal necrosis,apoptosis and significantly increased neuronal [Ca~2+ ]i.Compared with the damaged group,the cell survival rate increased significantly in all of the three groups of RCS-L1 1.0mg/L(P<0.01),0.1mg/L(P<0.01) and 0.01mg/L(P<0.05),and the percentage of neuronal necrosis decreased significanfly in all of the three groups of RCS-L1(P<0.001),while RCS-L1 at the concentration of 1.0mg/L and 0.1mg/L decreased the percentage of neuronal apoptosis and reduced neuronal [Ca~2+ ]i significanfly(P<0.05).Conclusion:It suggested that RCS-L1 was one of the representative brain protective components of Radix Codonopsis,and the neuronal protective mechanism of RCS-L1 might be the anti-apoptosis effect that might be related to decreasing the intracellular free calcium ion concentration.更多还原