摘要
目的研究重组人促红细胞生成素（rhEPO）在人视网膜色素上皮（RPE）细胞光化学损伤中的保护作用及其作用机制，为年龄相关性黄斑变性等眼部病变的药物治疗和预防提供理论依据。方法为实验研究。取成人ARPE-19细胞株传代培养的2～5代细胞建立光损伤模型，光照12h后再培养24h终止培养，采用噻唑蓝比色法和AnnexinV-FITC／PI流式双染法检测不同浓度的rhEPO干预治疗前后RPE细胞活性的变化及细胞凋亡的变化，采用酶联免疫吸附实验和免疫组织化学法定量检测不同含量rhEPO干预治疗前后RPE细胞caspase-3表达的变化，并添加AG490（Jak2激酶抑制剂）探讨重组人EPO对人RPE细胞光化学损伤的保护性作用和途径及其保护性机制。结果rhEPO可明显增强RPE细胞的活性，以40U／mLEPO组结果最明显；明显减少光化学损伤诱导的人RPE细胞的凋亡，以40U／mLrhEPO组结果最明显。在加入AG490组，rhEPO对细胞活性的增强作用和抑制凋亡作用均被阻止。结论rhEPO对人RPE细胞的光化学损伤有保护治疗作用，主要通过EPO的受体后保护作用机制起作用，即通过与受体结合，激活Jak2激酶的途径实现的。（中华眼科杂志，2008，44：50-55）
Objective The aim of this study was to assess the protection of recombinant human erythropoietin （rhEPO） in light-induced injures in human retinal pigment epithelial （RPE）cells. Methods It was a experimental study. Cultured human RPE cells were exposed to light of 8w 2000 ± 500 Lux for 12 hours. The 3-（4,5-dimethylthiazole-2yl ）-2,5-diphenyl tetrazolium bromide （MTF） cell viability assay were used to assess the effects of rhEPO in light-induced injury on human RPE cells. The effect of inhibiting apoptosis of rhEPO was detected by AnnexinV-flunorescein isothiocyanate/Propidium iodium labeling and flow cytometry. The enzyme linked immunosorbant assay （ELISA） and immunocytochemical staining were used to assess the expressions of caspase-3 treated by different doses of rhEPO in light-induced injury on human RPE cells and examine the protective mechanism of rhEPO by treatment with AG490 （ the special inhibitor of jak2）. Results There was a significant increase of inhibiting apoptosis in every rhEPO group, and cell viability was the highest in 40 U/ml rhEPO group, the value was 4.93 ± 1.45/ml. The decrease in expression of caspase-3 was the most obvious in 40 U/ml rhEPO group, in which the value was 0. 125 ± 0. 029 ng/ml. There was a significant increased effect on inhibiting apoptosis in every rhEPO group, and it was the most conspicuous in 40 U/ml rhEPO group. But these increased cell viability and effect on inhibiting apoptosis in rhEPO group were restrained by AG490, in which the value of apoptosis was 11.29 ± 2.11/ml and the density of caspase-3 increased to 0. 362 ± 0. 042 ng/ml. Conclusions It is suggested that rhEPO can protect human RPE cells from the light-induced injures. Its protective mechanism is principally mediated by the EPO-EPOR pathway, which subsequently leads to jak2 activation. （Chin J Ophtbalmol, 2008,44： 50-55）
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2008年第1期50-55,共6页
Chinese Journal of Ophthalmology
基金
国家自然科学基金资助项目（30572010）
关键词
色素上皮
眼
光刺激
红细胞生成素
重组
细胞凋亡
Pigment epithelium of eye
Photic stimulation
Erythropoietin, recombinant
Apoptosis
