【作者】 张壮； 韦颖； 闫彦芳； 牛福玲； 崔巍； 王硕仁； 孙塑伦； 朱陵群；

【Author】 ZHANG Zhuang~1,WEI Ying~1,YAN Yan-fang~1,et al (1.The Affiliated Dongzhimen Hospital of Beijing University of TCM,Beijing 100700,China; 2.Beijing University of TCM,Beijing 100700,China)

【机构】 北京中医药大学东直门医院； 北京中医药大学；

【摘要】 [目的]以原代培养的大鼠大脑星形胶质细胞进行缺氧缺糖/再给氧处理,观察党参总皂苷对星形胶质细胞损伤的保护作用。[方法]1d龄Wistar大鼠乳鼠大脑星形胶质细胞原代培养,含连二亚硫酸钠的无糖Earle’s液进行缺氧缺糖处理造模,四甲基偶氮唑蓝(MTT)法测定细胞存活率,Hoechst 33 342和PI原位双染法荧光显微镜观察细胞形态学变化和细胞坏死、凋亡率,比色法测定乳酸脱氢酶(LDH)漏出率,评价药效。[结果]1)细胞存活率(%):与模型组(52.2±7.3)比较党参总皂苷0.52 mg/L组(68.5±3.1)、0.005 2 mg/L组(64.3±5.1)均能显著提高细胞存活率(P<0.01);2)细胞坏死率(%):与模型组(34.0±3.2)比较党参总皂苷0.52mg/L组(18.0±2.3)、0.052mg/L组(11.3±2.7)和0.005 2 mg/L组(11.4±2.9)均能显著降低细胞坏死率(P<0.01);3)原位双染法细胞凋亡率(%):与模型组(10.6±4.7)比较党参总皂苷0.52 mg/L组(14.1±5.3)、0.052 mg/L组(10.1±6.0)和0.005 2 mg/L组(10.1±3.4)对细胞凋亡率均无显著影响(P>0.05);4)LDH漏出率(%):与模型组(20.9±3.7)比较党参总皂苷党参总皂苷0.52mg/L组(12.8±2.0)、0.052mg/L组(12.0±1.2)和0.005 2mg/L组(12.3±1.2)均能显著降低细胞LDH漏出率(P<0.01)。[结论]党参总皂苷对缺血再灌注损伤后星形胶质细胞的坏死具有显著抑制作用,但对凋亡过程无保护作用,提示党参总皂苷是党参治疗中风病急性期的主要效应成分。更多还原

【Abstract】 [Objective]To study the protective effect and its mechanism of the total saponin of Radix Codonopsis(RCTS) on the damage induced by hypoxia/hypoglycemia and reoxygenation in cultured rat primary astrocytes.[Method]The model rats of hypoxia/ hypoglycemia and reoxygenation induced damage was set up in vitro by sodium dithionate.The cells survival rate was detected by MTT assay.Morphological changes of astrocytes necrosis and apoptosis were observed by double stain of hoe33 342 and propidium iodide with fluorescence microscope.Lactate dehydrogenase(LDH) release was detected by chromometry.[Result]Hypoxia/ hypoglycemia cultures and reoxygenation induced astrocyte necrosis,apoptosis and significantly increased LDH release.Compared with the damaged group,the cell survival rate(%) increased significantly in the protective groups RCTS 0.52 mg/L(68.5±3.1) and RCTS 0.005 2 mg/L(64.3±5.1)(P<0.01).Meanwhile,both the percentage of astrocytes cells necrosis and LDH release decreased and in the protective groups of 0.52 mg/L(18.0±2.3 and 12.8±2.0),0.052 mg/L(11.3±2.7 and 12.0±1.2) and 0.005 2 mg/L (11.4±2.9 and 12.3±1.2)(P<0.01),but RCTS of different concentration might have no effect on the apoptosis of astrocytes cells (P>0.05).[Conclusion]It suggested that RCTS was one of the representative brain protective component groups of Radix Codonopsis,and the astrocytes cells protective mechanism of RCTS was the effect of anti-necrosis,decreasing LDH release.更多还原