摘要
目的:研究丹酚酸B对谷氨酸诱导PC12细胞兴奋毒的保护及作用机制。方法:以谷氨酸损伤PC12细胞24 h为模型,采用四甲基偶氮唑蓝法检测细胞存活率;LDH法检测乳酸脱氢酶的漏出率;AO/EB双染法荧光显微镜和PI单染流式细胞术检测细胞凋亡;流式细胞术检测细胞内活性氧的含量;Western blotting法检测细胞内Caspase-3蛋白的表达。结果:丹酚酸B可明显抑制谷氨酸诱导的PC12细胞的损伤,阻止细胞LDH的释放,在50～200μmol.L-1剂量呈一定的量效关系;同时,丹酚酸B明显降低谷氨酸诱导的活性Caspase-3蛋白的表达,抑制谷氨酸引起的ROS的累积,降低PC12细胞的凋亡率,在50～200μmol.L-1剂量呈量效相关性。结论:在一定剂量范围内,丹酚酸B对谷氨酸损伤的PC12细胞有保护作用,其保护的机制可能与丹酚酸B减少ROS的生成,阻止氧化损伤的发生,抑制Caspase-3途径依赖的凋亡相关。
Objective： To study the protective effect and mechanism of salvianolic acid B （ Sal B） on glutamate-induced excitotoxicity. Method： Glutamate-induced PC12 cell injury model was established to detect the cell survival rate by MTT, the leakage rate of lactic dehydrogeuases using LDH, and the cell apoptosis by using AO/EB double staining for fluorescence microscope and PI single staining flow cytometry which was also used to detect the content of intraceflular reactive oxygen species. The expression of Caspase-3 protein was also detected by the Western blotting method. Result： Sal B is proved to inhibit glutamate-induced PC12 cells from injury and prevent them from releasing LDH within the range from 50 p^mol ~ L- 1 to 200 ~mol ~ L-l. Meanwhile, Sal B has an effect on sig- nificantly reducing the expression of inhibit glutamate-induced active Caspase-3 protein, inhibiting accumulated glutamate-induced ROS and decreasing PC12 cell apoptosis rate within the range from 50μmol·L-1 to 200μmol·L-1. Conclusion： The study proves that Sal B prevented against glutamate-induced cell injury via inhibiting ROS formation and Caspase-3 pathway-dependent apoptosis in PC12 ceils.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2012年第3期353-357,共5页
China Journal of Chinese Materia Medica
基金
安徽高等学校省级自然科学研究项目(KJ2009B199Z)
