促红细胞生成素对大鼠视网膜Müller细胞低氧损伤的保护作用  

The protective effect of erythropoietin on cultured retinal Müller cells injured by hypoxia

作　　者：曲虹[1] 厉泉[1] 

机构地区：[1]山东省千佛山医院眼科,济南250014

出　　处：《中华临床医师杂志（电子版）》2015年第14期86-89,共4页Chinese Journal of Clinicians(Electronic Edition)

基　　金：山东省自然科学基金(ZR2011HL057;ZR2013HM028)

摘　　要：目的探讨促红细胞生成素对大鼠视网膜Müller细胞低氧损伤的保护作用。方法传代培养大鼠视网膜Müller细胞,分为N组(正常对照)、C50、C100、C150、C200、C300组(氯化钴浓度50,100,150,200,300μmol/L),MTT检测不同浓度氯化钴对Müller细胞活力的影响,确定制作低氧损伤的氯化钴浓度。然后再分组为N组(正常对照),C组(200μmol/L氯化钴),E1C组(1×104 IU/L rh EPO+200μmol/L氯化钴),E2C组(2×104 IU/L rh EPO+200μmol/L氯化钴),E4C组(4×104 IU/L rh EPO+200μmol/L氯化钴),MTT比色法比较五组视网膜Müller细胞活力的改变。Western blot检测各组细胞谷氨酰胺合成酶蛋白表达的情况。结果氯化钴浓度低于200μmol/L时,细胞活力不受氯化钴浓度的影响。从200μmol/L开始,细胞活力随氯化钴浓度增高而下降。E1C、E2C、E4C组细胞活力均高于C组,E4C组高于E2C组和E1C组,C组低于N组。Western blot检测谷氨酰胺合成酶表达结果显示,C组低于N组,EPO干预各组蛋白表达均高于C组,并随EPO浓度增高表达增高,但均低于N组。结论 EPO对视网膜Müller细胞低氧损伤有保护作用。Objective To investigate the protective effect of erythropoietin （EPO） on cultured retinal Muller cells injured by hypoxia. Methods Neonatal rats＇ retinal Muller cells of serial subcultivation were divided into N （normal control group） and Cs0, C100, C150, C200, C300 groups according to the different concentration of cobalt chloride （50, 100, 150, 200, 300 μmol/L）. MTT assay was applied to detect cell viability of every group. Then cultured cells were divided into N （normal control） group, C （200 μmol/L cobalt chloride） group, EIC （1 - 104 IU/L rhEPO＋200 μmol/L cobalt chloride） group, E2C （2× 10^4 IU/L rhEPO＋200μmol/L cobalt chloride）group, E4C （4×10^4 IU/L rhEPO＋200 μmol/L cobalt chloride）. MTT assay was applied to detect cells viability in five groups. The expression of glutamine synthetase was detected with Western blot method. Results MTT assay indicated that there was no change of cell viability when the concentrations of cobalt chloride were lower than 200 μmol/L, then, the cell viability decreased along with the concentration of cobalt chloride increased. Cell viability of E1C, E2C and E4C group was higher than that of C group, while, E4C group was higher than those of E2C and E1C group. The expressions of glutamine synthetase in E4C, E2C and E1C group were higher than that in C group which was low compared with N group. Conclusion EPO pretreatment could effectively protect the cultured retinal M/iller cells from injuries induced by hypoxia.

关 键 词：细胞低氧 谷氨酰胺连接酶 视网膜 Mfiller细胞 重组人促红细胞生成素: 氯化钴 

分 类 号：R774.1[医药卫生—眼科]