【作者】 颜涛；

【导师】 严奉祥；

【作者基本信息】 南华大学 ， 药理学， 2006， 硕士

【摘要】 目的：观察迷迭香酸(Rosmarinic acid，RosA)对谷氨酸(Glutamic acid，Glu)诱导PC12细胞凋亡的作用，初步探讨迷迭香酸抗谷氨酸诱导PC12细胞凋亡的机制。 方法：以谷氨酸损伤的PC12细胞为模型，用普通光镜和吖啶橙(AO)荧光染色观察谷氨酸对PC12细胞形态和核形态的影响；用四甲基偶氮唑盐(MTT)法观察迷迭香酸对PC12细胞存活率的量-效关系和时-效关系；用碘化丙啶(Propidium iodide，PI)染色流式细胞术检测迷迭香酸对PC12细胞凋亡率的效应；用RT-PCR、间接免疫荧光法和Western blot分别检测迷迭香酸对PC12细胞Bcl-xl和Bax mRNA含量和蛋白质表达的影响。 结果：用不同剂量谷氨酸(1、5、10、12.5、15、20mmol／L)孵育PC12细胞24h后，该细胞存活率随药物浓度的增加依次下降，12.5mmol／L谷氨酸孵育细胞24h后，其细胞形态发生了明显的改变，并出现细胞核浓缩、碎裂和凋亡小体等凋亡特征；Bcl-xl表达下调，Bax表达上调。用100μmol／L迷迭香酸预处理细胞1h后，再用12.5mmol／L谷氨酸孵育细胞24h，细胞凋亡率由21.63％±0.25％下降至4.23％±1.32％(P＜0.01)，Bcl-xl表达上调(P＜0.01)，Bax表达下调(P＜0.05)。加Bcl-xl蛋白质阻断剂HA14-1(40μmol／L)后，迷迭香酸抗谷氨酸诱导的PC12细胞凋亡的作用完全被取消(11.62％±0.65％ vs 2.55％±0.13％)，其凋亡率甚至比单独用Glu处理组还高(11.62％更多还原

【Abstract】 Objective: The study was to investigate the effects of rosmarinic acid on the PC12 cells apoptosis inducd by glutamic acid and explore the involved underling molecular mechanism.Methods: With the model of PC12 cell injury induced by glutamic acid, the cell shape and nuclear morphology were observed by light microscope and nuclear staining with acridine orange (AO), respectively. The relationships both dose-effect and time-effect of rosmarinic acid on survival rate of the PC12 cells were determined by 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The effect of rosmarinic acid on apoptotic rate of PC12 cells was determined by flow cytometry analysis dyed with propidium iodide (PI). RT-PCR, indirect immunofluorescence and western blot were used to detect the effect of rosmarinic acid on Bcl-xl, Bax mRNA and protein expression in PC12 cells, respectively.Results: After PC12 cells incubated with deffrent dose of glutamic acid (1, 5, 10, 12.5, 15, 20mmol/L) for 24 hours, the survival rate of cells was reduced in dose- dependent manner. While PC12 cells were pretreated of 12.5 mmol/L glutamic acid for 24 hours, the PC12 cells were evidently changed in shape, presented clitypical morphological更多还原