促红细胞生成素预处理对视网膜神经元缺氧性损伤的保护作用  

Protective effects of recombinant human erythropoietin(EPO) on rat retinal neurons hypoxia injuries

作　　者：刘夫玲[1] 牛膺筠[1] 王红云[1] 赵颖[1] 曲虹[1] 

机构地区：[1]青岛大学医学院附属医院眼科,山东青岛266003

出　　处：《中国药理学通报》2007年第2期228-233,共6页Chinese Pharmacological Bulletin

基　　金：山东省科技攻关项目基金资助(No2004GGB14112)

摘　　要：目的研究缺氧对培养大鼠视网膜神经元的影响及促红细胞生成素(erythropoietin,EPO)预处理的保护作用。方法取体外原代培养3d的大鼠视网膜神经元,利用1mmol.L-1的连二亚硫酸钠消除培养基中的氧合并培养基质缺糖持续6h,导致神经元缺氧性损伤。采用免疫组化染色技术检测了EPO受体在原代培养视网膜神经元的表达,乳酸脱氢酶(LDH)释放率作为评价损伤的指标,TUNEL染色检测视网膜神经元的凋亡,并进行形态学观察。结果原代培养的大鼠视网膜神经元正常下可见EPOR的弱阳性表达,表达部位定位于神经元的胞体和突起,缺氧后EPOR的表达明显增加;神经元LDH释放率和凋亡百分率增加;在损伤前加入浓度2.5～40kU.L-1的rhEPO均可有效抑制LDH释放(P<0.05)。TUNEL结果显示rhEPO预处理能抑制神经元的凋亡,明显改善细胞缺氧性损伤的形态学变化。结论EPO预处理对神经元的缺氧损伤具有保护作用,且具有浓度和时间依赖性。Aim To study the protective effects of recombinant human erythropoietin （ rhEPO ） pretreatment on rat retinal neurons injuries induced by defect of oxygenation/glucose in vitro. Methods The rat retinal neurons were cultured for 3 d, and placed in the environment of Na2 S2O4 （ 1.0 mmol· L^-1 ） and glucose defeetion for 6 h to induce neuronal cells hypoxia injury. The expression of erythropoietin receptor （EPOR） was analyzed by immunohistochemistry, the releasing quantity of LDH was determined as index of neuronal cells injury. Terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling （TUNEL） assay demonstrated the neuron apoptosis induced by hypoxia. After being treated with rhEPO, the protective effects of the drug on neuronal cells injury and apoptosis were observed. Results Cultured rat cortical neurons expressed EPOR faintly, located in the neuron body and neurites. Induction of hypoxia caused a dramatic increase of EPOR expression, the releasing quantity of LDH and apoptosis increased markedly after hypoxia injuries. Being treated with rhEPO （2. 5-40 kU ·L^-1 ） the hypoxia injuries of neuronal cells relieved effectively. TUNEL assay indicated the apoptosis lowered markedly after rhEPO pretreatment. Conclusion EPO pretreatment could effectively protect the neuronal cells from injuries induced by hypoxia, which was positively related to the time and dosage.

关 键 词：促红细胞生成素(EPO) 缺氧 视网膜 神经元 细胞培养 

分 类 号：R-332[医药卫生] R322.91