大鼠缺氧神经元细胞中促红细胞生成素及其mRNA的表达  

Erythropoietinprotein and its mRNA expression in rats cortical neurons following hypoxia

作　　者：李俊[1] 陶陶[2] 邹哲华[3] 徐坚[4] 罗开俭[5] 刘智[6] 

机构地区：[1] 华中科技大学同济医学院附属普爱医院神经内科 [2] 贵阳医学院附属人民医院康复医学科, 550002 [3] 河北省秦皇岛市第一医院老年病科 [4] 贵阳医学院附属医院神经内科 [5] 贵阳医学院附属医院急诊科 [6] 贵阳医学院附属医院药剂科

出　　处：《中华神经科杂志》2014年第4期254-257,共4页Chinese Journal of Neurology

基　　金：贵州省科学技术基金资助项目（黔科合J字[2007]2096号）

摘　　要：目的 观察大鼠大脑皮质神经元细胞缺氧时促红细胞生成素（erythropoietin,EPO）及其mRNA表达的变化,探讨内源性EPO对缺氧神经元细胞的保护作用.方法 制备体外培养大鼠大脑皮质神经元细胞缺氧模型,分为正常对照组；缺氧12、24、48、72 h组.利用免疫组织化学、逆转录聚合酶链反应及蛋白质印迹观察细胞中EPO及其mRNA的表达,并在变化过程中观察培养液中乳酸脱氢酶来评估神经元细胞的活性.结果 免疫组织化学、逆转录聚合酶链反应及蛋白印迹检测显示：缺氧组中EPO（20.79±2.98）及其mRNA（0.78±0.05）在12 h已有基本表达,与对照组（EPO：17.12±1.99；mRNA：0.39±0.05）比较,差异均有统计学意义（t=2.51,P＜0.05；t=13.51,P＜0.01）；且48 h达高峰（EPO：28.88±3.41,mRNA：1.45±0.07）,与对照组比较,差异有统计学意义（t =7.29,P＜0.叭；=33.24,P＜0.01）；神经元细胞活性也最强.乳酸脱氢酶活性72 h后表达明显下降,各时间组之间比较差异同样具有统计学意义.结论 神经元细胞缺氧后使细胞内EPO及EPO mRNA的表达增强,且可增强神经元细胞的活性.提示EPO参与了神经元细胞缺氧的发生发展过程,在神经元细胞缺氧的过程中起重要作用.Objective To observe erythropoietin （EPO） and its mRNA expression changes in rats cortical neurons when suffering hypoxia and investigate the endogenous EPO protective effect of hypoxia neuronal cells.Methods Cultured cortical neurons were prepared from hypoxia rats and divided into control,hypoxia 12,24,48,72 h group.Using immunohistochemistry,reverse transcriptase polymerase chain reaction （RT-PCR） and Western blot,we observed EPO and its mRNA expression in cells,and also observe the culture medium containing lactate dehydrogenase to evaluate the activity of neurons in the whole process.Results Immunohistochemistry,RT-PCR and Western blot analysis showed：the EPO （20.79 ± 2.98） and its mRNA （0.78 ± 0.05） at 12 h had a basic expression in hypoxia group,compared with the control group （EPO：17.12 ± 1.99; mRNA：0.39 ± 0.05）,and the difference was statistically significant （t =2.51,P 〈 0.05 ; t =13.51,P 〈 0.01） ; the strongest expression was observed at 48 h （EPO：28.88 ± 3.41,mRNA：1.45 ± 0.07）,the difference was statistically significant （t =7.29,P 〈 0.01 ; t =33.24,P 〈 0.01） ; and neuronal activity was strongest.Lactate dehydrogenase activity was significantly decreased after hypoxia 72 h,and also a statistically significant difference was found between the groups at each point.Conclusions The EPO and EPO mRNA expression are increased after hypoxia in neuron cells,and may enhance the activity of neurons.Our study suggests that EPO might be involved in the development process of neuronal hypoxia and play an important role in neuronal hypoxia process.

关 键 词：红细胞生成素 大脑皮质 神经元 细胞低氧 

分 类 号：R741[医药卫生—神经病学与精神病学]