鱼藤酮诱发PC12细胞内质网应激的钙机制和超微结构的变化  

Calcium mechanism of endoplasmic reticulum stress and the change of ultrastructures induced by rotenone in PC12 cells

作　　者：赵黔鲁[1] 柴锡庆[1] 

机构地区：[1]河北医科大学研究生院

出　　处：《中华神经科杂志》2005年第6期363-366,共4页Chinese Journal of Neurology

摘　　要：目的探讨鱼藤酮诱发内质网(endoplasmicreticulum,ER)应激的钙离子机制及其超微结构的变化。方法应用流式细胞仪(flowcytometry,FCM)检测鱼藤酮对体外培养大鼠肾上腺嗜铬细胞瘤(PC12)细胞株活性氧的影响,应用激光扫描共聚焦显微镜观察细胞内钙离子变化及透射电镜观察超微结构。结果0·1、1·0、2·0和3·0μmol/L浓度鱼藤酮诱导细胞内活性氧的生成,荧光指数(FI)值分别为1·55±0·17、2·16±0·10、1·77±0·20和1·41±0·12,相同浓度鱼藤酮所致细胞内钙离子荧光强度变换值分别为0·6029±0·0685、1·0902±0·1127、0·7479±0·0820和0·5614±0·0870,分别与对照组比较差异有统计学意义(P<0·01)。当鱼藤酮浓度大于1·0μmol/L时,其作用呈剂量依赖性递减。预先用超氧化物歧化酶(SOD)1200U/ml干预24h,可有效减弱1·0μmol/L鱼藤酮引发的细胞内钙离子升高(P<0·01)。电镜观察到滑面内质网大量增生,粗面内质网轻、中度扩张及脱颗粒。结论鱼藤酮可通过活性氧途径耗竭ER腔钙离子,诱发ER应激,并引起其超微结构的相应变化。Objective To investigate the calcium mechanism of endoplasmic reticulum stress and the changes of ultrastructure induced by rotenone. Methods Rotenone induced reactive oxygen species(ROS) production in PC12 cells were measured by using flow cytometry(FCM). Relative changes of intracellular calcium concentration were monitored by use of laser scanning confocal microscope. The ultrastructures were observed under transmitting electronic microscope. Results Treatment with different concentrations of rotenone ( 0.1 μmol/L,1.0 μmol/L,2.0 μmol/L and 3.0 μmol/L )increased the ROS generation in PC12 cells, and fluorescence intensity (FI) value was shown as 1.55±0.17, 2.16±0.10, 1.77±0.20 and 1.41±0.12, respectively. The same concentration of rotenone resulted in the elevation of intracellular calcium,the relative changes of fluorescence intensity about 0.6029±0.0685,1.0902±0.1127,0.7479±0.0820 and 0.5614±0.0870. There were significant differences as compared with those in control group (P<0.01). When rotenone concentrations exceeded 1.0 μmol/L, the effects were decreased in a dose-dependent manner. Preincubation with superoxide dismutase (SOD) 1200 U/ml for 24 hours significantly attenuated the elevation of intracellular calcium induced by rotenone 1.0 μmol/L (P<0.01). Ultrastructural alterations showed a smooth endoplasmic reticulum in great deal of hyperplasia and a rough endoplasmic reticulum enlargement as well as degranulation in mild and moderate degrees. Conclusions Rotenone might deplete Ca~ 2+ of the endoplasmic reticulum lumen by ROS pathway inducing endoplasmic reticulum stress. It might also cause ultrastructural alterations of endoplasmic reticulum.

关 键 词：超微结构 鱼藤酮 PC12细胞 内质网应激 诱发 CYTOMETRY 肾上腺嗜铬细胞瘤 细胞内钙离子变化 mol/L 钙离子荧光强度 超氧化物歧化酶 透射电镜观察 细胞内活性氧 钙离子机制 流式细胞仪 剂量依赖性 钙离子升高 滑面内质网 

分 类 号：R114[医药卫生—卫生毒理学]