天麻素通过抑制细胞凋亡保护谷氨酸诱导的PC12细胞损伤

目的：研究天麻素（Gastrodin）对谷氨酸诱导的大鼠肾上腺嗜铬细胞瘤PC12细胞损伤的影响及可能机制。方法：以谷氨酸建立体外培养PC12细胞损伤模型并采用MTT比色法测定细胞存活率;AO/EB双染法经荧光显微镜观察细胞凋亡形态;采用流式细胞术检测细胞内活性氧含量以及Annexin V/PI染色后的细胞凋亡率;Western blot法检测细胞内Caspase-3蛋白表达。结果：天麻素可明显抑制谷氨酸诱导的PC12细胞凋亡,在0.1～10μmol/L剂量呈一定的量效关系;同时,天麻素可明显抑制谷氨酸引起的活性氧（ROS）的累积,降低谷氨酸诱导的活性Caspase-3蛋白的表达,降低PC12细胞的凋亡率,在0.1～10μmol/L剂量呈量效相关性。结论：在一定剂量范围内,天麻素对谷氨酸损伤的PC12细胞具有保护作用,其机制可能与减少ROS的生成,阻止氧化损伤的发生,抑制Caspase-3途径依赖的细胞凋亡相关。

Protection of gastrodin on PC12 cell injury induced by glutamate via inhibition of cell apoptosis

AIM： To study the protection and the molecular mechanisms of gastrodin on PC12 cells injury induced by glutamate.METHODS： The injury model was established by treating PC12 cells with glutamate in vitro and the cell activity was analyzed by MTT assay.Cell apoptosis morphological changes were detected by AO/EB staining with fluorescence microscope.The flow cytometry（FCM） was used to evaluate the level of intracellular reactive oxygen species（ROS） and to examine cell apoptosis rate via Annexin V-FITC/PI double staining.The expression of caspase-3 protein was determined by Western blot methods.RESULTS：It was indicated that gastrodin can apparently inhibit PC12 cell apoptosis induced by glutamate,it presented a dose-response relationship of the dosage 0.1-10 μmol/L.Gastrodin can also reduce ROS level,drop the expression of cleaved caspase-3 protein and decrease cell apoptosis rate,and gastrodin of 0.1-10 μmol/L presented a dose-response relationship.CONCLUSION： Gastrodin has a protective effect on apoptosis of PC12 cell within a certain dosage range,the protective mechanism of gastrodin on PC12 cell injury induced by glutamate may be related to gastrodin reducing the releasing of ROS,preventing occurrence of oxidative damage and thereby inhibiting caspase-3 pathway-dependent apoptosis in PC12 cell.