PTEN表达下调对神经元胞内游离钙离子和神经元凋亡的研究  

The mechanism of PTEN down-regulation on neuron intracellular free calcium and apoptosis

作　　者：陈力学[1] 刘宝松[2] 康格非[1] 姜利人[2] 

机构地区：[1]重庆医科大学检验系临床生化室,重庆400016 [2]第三军医大学大坪医院野战外科研究所三室,重庆400042

出　　处：《重庆医科大学学报》2006年第4期505-508,共4页Journal of Chongqing Medical University

基　　金：重庆市科委应用基础研究资助项目(8107)

摘　　要：目的:探讨神经元缺血缺氧损伤再灌后,下调PTEN在阻断Ca2+内流及凋亡调控方面的保护机制。方法:建立神经元缺糖缺氧(oxygen-glucose deprivation,OGD)损伤模型,转染shRNAspten-GFP质粒,用Western blotting检测PTEN水平,Fura2-AM法测定胞内Ca2+浓度,流式细胞术和AO/EB检测神经元凋亡。结果:与正常组相比OGD后细胞凋亡率和胞内Ca2+浓度显著升高(P<0.05);与对照组相比经shRNAspten干预后胞内Ca2+浓度和细胞凋亡率明显降低(P<0.05)。结论:shRNAspten-GFP能成功转染到神经元中,PTEN表达下调可阻断Ca2+内流并对缺糖缺氧诱导的细胞凋亡有拮抗作用,从而达到神经元保护的目的。Objective： To investigate the regulation of PTEN on apoptosis and the active roles in blocking Ca^2＋ influx and neuroprotection. Methods： In the OGD model with anaerobic gas mixture and deoxygenated, glucose-free extracellular solution, shRNAspten-GFP and unrelated control plasmid PshGFP transfection were done respectively. Intracellular free calcium concentration were measured with Fura-2 AM fluorescent indicator, the expression of PTEN were analyzed by immunoblotting（blot） and neuron apoptosis were detected by AO/EB and Flow Cytometry. Results： Apoptosis rate and calcium concentration after OGD was significantly higher than that of normal group （ P〈0.05 ）. But intracellular free calcium concentration and apoptosis with shRNAspten-GFP treatment after OGD was significantly lower than that in OGD group and PshGFP group （P〈0.05）. Conclusion： shRNAspten-GFP plasmid can been expressed successfully in cultured neurons. PTEN down-regulation could effectively suppress apoptosis occurrence induced by OGD and the increase of intracellular free calcium concentration, and then protect neuron from anoxia injury.

关 键 词：PTEN shRNAspten-GFP质粒 游离钙 凋亡 神经元培养 

分 类 号：R743[医药卫生—神经病学与精神病学]