氧化槐定碱对新生大鼠海马神经元氧糖剥夺再灌注损伤后细胞凋亡的影响  

Anti-apoptotic effects of oxysophoridine following oxygen-glucose deprivation and reperfusion in neonatal rat primary cultured hippocampal neurons

作　　者：周茹[1] 郝银菊[1] 姚婉霞[1] 杨文莉[1] 杜娟[1] 余建强[1] 

机构地区：[1]宁夏医科大学药学院药理学系,银川750004

出　　处：《中药药理与临床》2014年第6期32-35,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：宁夏留学人员创新创业个人项目择优资助项目

摘　　要：目的:研究氧化槐定碱对原代培养新生大鼠海马神经元氧糖剥夺再灌注损伤后细胞调亡的影响。方法:以原代培养的新生大鼠海马神经元为研究对象建立氧糖剥夺再灌注损伤模型。Hoechst 33342和TUNEL染色法检测海马神经细胞凋亡,透射电子显微镜术观察凋亡细胞形态学变化,化学比色法测定海马神经细胞Caspase-3、Caspase-9和Caspase-8的活性,免疫蛋白印迹技术和实时荧光定量PCR技术检测凋亡相关因子Cytochrome C、Caspase-3、Bcl-2、Bax和PARP-1的蛋白和mRNA的表达。结果:与正常组比较,损伤组Hoechst 33342和TUNEL染色法显示神经细胞凋亡率明显升高;Caspase-3、Caspase-9和Caspase-8活性显著提高;Cytochrome C、Caspase-3、Bax和PARP-1蛋白和mRNA水平明显上调,Bcl-2蛋白和mRNA水平明显下调,Bcl-2/Bax比率也明显下调。与损伤组比较,氧化槐定碱治疗组(20、5、1.25mg/L)可显著改善氧糖剥夺再灌注损伤所致的神经细胞凋亡的形态学变化,并降低神经细胞凋亡率;明显抑制Caspase-3、Caspase-9和Caspase-8的活性。氧化槐定碱治疗组(20mg/L)可明显抑制Cytochrome C、Caspase-3、Bax和PARP-1蛋白和mRNA表达,增强Bcl-2蛋白和mRNA的表达,使Bcl-2/Bax比率上升。结论:氧化槐定碱通过抗凋亡作用对氧糖剥夺再灌注损伤的大鼠海马神经细胞发挥保护作用。Objective： To study the protective effects of oxysophoridine on hippocampal neurons injured by oxygen-glucose deprivation/reperfusion （OGD/RP） in vitro and elucidated the related mechanisms. Methods： Cultured hippocampal neurons exposed to oxygen-glucose deprivation （OGD） for 2h followed by a 24h re-oxygenation were used as an in vitro model of ischemia and reperfusion. The neuron apoptosis was evalua- ted by Hoechst 33342 staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay （TUNEL）. Morphological change in the neurons was examined using a Transmission Electron Microscope （ TEM or EM ）. To evaluate neuron apoptosis, Caspase-3, -9, and -8 activities were measured using assay kits by chemical colorimetry. Western blotting assay and Quantitative real-time PCR assay were used to evaluate the release of cytochrome c, protein and mRNA expression of Caspase-3, Bcl-2 , Bax and PARP-1. Results： Compared with the OGD/RP group, treatment with OSR （20,5,1.25 mg/L） significantly attenuated neuronal damage, with evidence of decreased cell apop- tosis and decreased cell morphologlc impairment. Compared with the OGD/RP group, treatment with OSR （20, 5, 1.25 mg/L） signiiricandy attenuated these increase of Caspase-3, Caspase-9 and Caspase-8 activities. Compared with the OGD/RP group, treatment with OSR （20mg/ L） coald effectively down regulate the expression oir Cytochrome C, Caspase-3, Bax and PARP-land in protein level, induce an increase of Bcl-2in protein level, Bcl-2/Bax ratio also was increased. Compared with the OGD/RP group, treatment with OSR （20mg/L） could signifi- cantly down regulate the expression of Cytochrome C, Caspase-3, Bax and PARP-1 in protein level, induce an increase of Bcl-2 in mRNA level. Conclusion： Oxysophoridine affords protective effects may be mediated by its action on the downstream Caspase-3, Caspase-9, and Caspase-8 activities,suppression of mitochondrial Cytochrome C release into cytosol and PARP-1, and up-regalation of Bc

关 键 词：氧化槐定碱 氧糖剥夺再灌注损伤 抗凋亡 

分 类 号：R285[医药卫生—中药学]