补充硒蛋氨酸促进低氧/复氧损伤神经元存活作用的体外研究  

STUDY ON NEURON SURVIVAL DURING HYPOXIA/REOXYGENATION AFTER SELENOMETHIONINE SUPPLEMENT IN VITRO

作　　者：窦贺荣[1] 李贤慧[2] 张新昌[2] 李积胜[3] 何冰[3] 

机构地区：[1]武警后勤学院实验动物中心 [2]武警后勤学院生化与分生教研室 [3]武警后勤学院天津市职业与环境危害生物标志重点实验室,天津300162

出　　处：《营养学报》2013年第4期369-373,共5页Acta Nutrimenta Sinica

基　　金：天津市应用基础及前沿技术面上项目(No.11JCYBJC11600)

摘　　要：目的研究低氧/复氧损伤时补充硒蛋氨酸(selenomethionine,SeMet)对神经元存活的影响及可能机制分析。方法原代培养大鼠脑皮质神经元低氧1h复氧6h,低氧/复氧同时加入SeMet。实验分为正常对照组、单纯低氧/复氧组、低氧/复氧补充SeMet 1、1.5、2μmol/L组。用CCK-8试剂检测细胞活力,Annexin V-FITC/PI双标记检测细胞凋亡,细胞总谷胱甘肽过氧化物酶(GPx)活力用试剂盒分析,Western–Blotting进一步分析谷胱甘肽过氧化物酶-1(GPx-1)和磷脂氢谷胱甘肽过氧化物酶(GPx-4)表达。结果低氧1h/复氧6h后神经元损伤明显,与对照组相比细胞活力降低至46%,凋亡率达到70.5%。补充SeMet可使神经元细胞活力下降减轻、神经元凋亡减少。GPx活力分析和Western–Blotting结果表明低氧/复氧时补充SeMet可使细胞总GPx活力增加,GPx-1和GPx-4,特别是GPx-4蛋白量明显增加。结论神经元低氧/复氧损伤时补充SeMet可增加GPx-1和GPx-4的表达进而促进神经元存活。Objective To investigate the effects of selenomethionine supplement on neuron survival during hypoxia/reoxygenation. Methods Cortical ueurons were cultured and exposed to hypoxia for lh and followed by reoxygenation for 6h. Experiments were performed on five groups： normal control group, hypoxia/reoxygenation group, selenomethionine 1.0, 1.5, 2 gmol/L groups. Cell viability was measured by CCK-8. Apoptosis was detected by using FITC-Annexin V/PI double staining. Glutathione peroxidase （GPx） activity was evaluated by GPx activity assay kit. The expression of GPx-1 and GPx-4 were analyzed by Western blotting. Results Neurons exposed to bypoxia for lh and followed by reoxygenation for 6h were injured markedly. Cell viability declined to 46% and the cell apoptosis increased to 70.5% compared with control. Selenomethionine supplement recovered the cell viability and prevented the cell ap0ptosis. GPx activity and Western blot assay showed that selenomethionine supplement not only increased total cellular GPx activity but also enhanced the expression of GPx-1 and GPx-4. Conclusion Selenomethionine supplement improves neuron survival during hypoxia/reoxygenation by enhancing the expression of GPx-1 and GPx-4.

关 键 词：硒蛋氨酸 神经元 低氧 复氧 谷胱甘肽过氧化物酶 体外 

分 类 号：R151[医药卫生—营养与食品卫生学]