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作 者:陈宏[1] 陈建宗[1] 厚荣荣[1] 杨爱莉[2] 康小刚[1] 张影[1] 李源莉[1] 李海龙[1]
机构地区:[1]第四军医大学西京医院中医药研究中心中西医结合老年脑病研究室,陕西西安710033 [2]第四军医大学西京医院老年病科,陕西西安710033
出 处:《中国药理学与毒理学杂志》2007年第3期190-196,共7页Chinese Journal of Pharmacology and Toxicology
摘 要:目的探讨红景天苷(salidroside)的神经保护作用及其可能的作用机制。方法采用百草枯(PQ)诱导的具有多巴胺神经元特性的PC12细胞凋亡作为帕金森病的体外模型。实验分对照组、PQ诱导组和红景天苷3个浓度(终浓度10,20和30μmol.L-1)预处理组。用MTT法测定细胞存活率,流式细胞术和Hoechst33258染色法测定细胞凋亡,比色法检测半胱氨酸天冬氨酸蛋白酶(caspase)-3活性,免疫组化法检测细胞色素c(Cytc)的释放和Bcl-2蛋白的表达。结果PQ(800μmol.L-1)作用于PC12细胞24h后,与正常对照组比较,细胞存活率降低,细胞染色质固缩,细胞核呈致密浓染,细胞凋亡百分率升高;红景天苷3个浓度预处理后,细胞存活率增加,细胞核凝聚明显减少,细胞凋亡率降低,且具有量-效关系。另外,PQ可使PC12细胞caspase-3活性增强,Cytc的释放增加,Bcl-2的表达代偿性升高;红景天苷预处理后,PC12细胞增高的caspase-3活性和Cytc的释放明显降低,Bcl-2的表达进一步升高。结论红景天苷对PQ诱导的细胞凋亡具有浓度依赖性的抑制作用,其作用机制可能是促进Bcl-2的表达,抑制Cytc的释放和caspase-3的激活,提示红景天苷可能具有神经保护作用。AIM To study the neuroprotective effect and its related mechanisms of salidroside. METHODS Paraquat ( PQ ) -induced apoptosis in PC12 cells, which having the characteristics of dopaminergic neuron, as the model of Parkinson disease in vitro. Cells were divided into control group, PQ group and 3 groups pretreated with salidroside of final concent.ration 10, 20 and 30 μmol·L^-1. The cell viability was measured with MTT assay. The apoptosis was monitored by using flow cytometry and Hoechst 33258 staining. Caspase-3 activity was detected with colorimetry, and the expressions of cytochrome c (Cyt c) and Bcl-2 were detected by using immunostaining assay. RESULTS Compared with control group, after exposure of PQ ( 800 μmol·L^-1 ) for 24 h, the viability of PC12 cells was decreased, chromatin was condensed, cell nuclei exhibited the dense fluorescence and apoptosis ratio was elevated. While after pretreatment with salidroside 10, 20 and 30 μmol·L^-1, resepectively, cell viability was increased, nuclear condensation was significantly reduced and apoptosis ratio was lowered. Moreover, this effect was concentration-dependent. In addition, in PQ-treated PC12 cells, caspase-3 activity and Cyt c release were increased, and compensatory expression of Bcl-2 was enhanced. Pretreated with salidroside, caspase-3 activity and Cyt c release were reduced obviously, and Bcl-2 expression elevated further. CONCLUSION Salidroside inhibits PC12 cell apoptosis induced by PQ in a concentration-dependent manner. The mechanisms of this effect include increasing the expression of Bcl-2, inhibiting the release of Cyt c and decreasing the activation of caspase-3, which suggests that salidroside has neuroprotective effect.
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