| 引用本文： |
-
雇利敏,张敬法,徐华,等.人源促红细胞生成素在Müller细胞中拮抗谷氨酸毒性作用的研究[J].同济大学学报（医学版）,2013,34(1):1-7. [点击复制]
- GU Li-min,ZHANG Jing-fa,XU Hua,et al.Erythropoietin protects Müller cells through maintaining its expression of glutamate-aspartate transporter[J].同济大学学报（医学版）,2013,34(1):1-7. [点击复制]
|
|
| 本文已被：浏览 406次 下载 384次 |
码上扫一扫！ |
| 人源促红细胞生成素在Müller细胞中拮抗谷氨酸毒性作用的研究 |
| 雇利敏1,2,张敬法2,徐华1,2,吕立夏2,李维业3,徐国彤2 |
|
|
| (1.同济大学生命科学与技术学院,上海200092 ; 2.同济大学附属第十人民医院眼科,上海200072;3.美MDrexel大学医学院眼科，费城19129) |
|
| 摘要: |
| 目的观察高浓度谷氨酸刺激条件下Mtiller细胞的变化以及促红细胞生成素（erythropoietin，EPO）的保护作用是否与维持Mtiller细胞的谷氨酸转运功能相关。方法用M1Tr法检测不同浓度谷氨酸处理大鼠原代视网膜Mtiller细胞和大鼠视网膜Miiller细胞系rMC一1所引起的细胞活力变化。选择能显著降低细胞活力的最低谷氨酸剂量建立细胞损伤模型，并研究不同浓度EPO干预后细胞活力的变化。TUNEL（terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling）法检测各组细胞凋亡情况，Western印迹法检测谷氨酸转运体GLSAT在蛋白水平的变化。结果无论是在原代大鼠Mtiller细胞还是rMC-1细胞系中，谷氨酸的细胞毒性作用均呈剂量依赖趋势。原代Miiller细胞中，12mmol/L谷氨酸使细胞活力降低15.5％，10mmol/L谷氨酸使rMC-1细胞活力降低15.6％；此时细胞凋亡明显增加，GLAST表达降低。0.2U/ml和0．5U/mlEPO分别对原代Mfiller细胞和rMC-1细胞的保护作用最佳，细胞凋亡数量显著减少，并防止了GLAST蛋白水平的降低。结论体外高浓度谷氨酸可损伤Mtiller细胞，EPO可以通过维持谷氨酸转运体GLAST水平等机制维持Mfiller对谷氨酸的正常摄取、抑制凋亡发生。 |
| 关键词: 促红细胞生成素 谷氨酸 Mtiller细胞 细胞凋亡 谷氨酸转运体 |
| DOI：10.3969/j.issn1008-0392.2013.01.001 |
|
| 基金项目:国家自然科学基金(81000383) |
|
| Erythropoietin protects Müller cells through maintaining its expression of glutamate-aspartate transporter |
| GU Li-min1,2,ZHANG Jing-fa2,XU Hua1,2,LI Li-xia2,LI Wei-ye3,XU Guo-Tong2 |
| (1.Tongji University School of Life Science and Technology,Shanghai 200092,China; 2.Dept.of Ophthalmology,Tenth People’s Hospital,Tongji University,Shanghai 200072,China;;3.Dept.of Ophthalmology,Drexel University College of Medicine,Philadelphia 19129,USA) |
| Abstract: |
| Objective To determine the mechanism of the protective effect of erythropoietin （EPO） on Miiller cells, with special focus on its effects on expression of glutamate-aspartate transporter （GLAST） under glutamate stress. Methods Rat retinal Mtiller cell line （rMC-1） and primary Miiller cells were employed in this study. MTT assay was used to exam the effects of different dosage of glutamate on viability of rMC-1 and primary MUller cells, and to evaluate EPO＇s protection under glutamate stress. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） method was used to detect cell apoptosis. GLAST level was studied with Western blotting method. Results Glutamate toxicity presented a dose-dependent manner on both primary Mtiller cells and rMC- 1 cells; 12 mmol/L glutamate caused 15.5% of the primary Mfiller cells to die and 10 mmol/L glutamate caused similar death rate in rMC-1 cells. TUNEL staining examination confirmed such cell death by demonstrating the increased apoptosis in both cell types and Western blotting showed a decreased GLAST expression in glutamate-treated cells. EPO （0.2 U/ml ） exerted its optimal protection on primary MUller cell against apoptosis induced by 12 mmol/L glutamate and also maintained GLAST at normal level. Similarly, rMC-1 cells were protected by EPO but had a better response to 0.5 U/ml EPO. Conclusion The present study demonstrates that EPO might protect Mtiller cells by maintaining their normal GLAST expression so that the glutamate was uptake into the MUller cells. |
| Key words: erythropoietin glutamate Mtiller cell apoptosis glutamate transporter |