重组人促红细胞生成素对离体培养视网膜神经节细胞的影响  

Effect of recombinant human erythropoietin on retinal ganglion cells cultured in vitro

作　　者：牛建军[1] 曾玉晓[1] 王一[1] 阴正勤[1] 王仕军[1] 陈中山[1] 刘康[1] 

机构地区：[1]第三军医大学西南眼科医院

出　　处：《眼科新进展》2007年第3期188-192,共5页Recent Advances in Ophthalmology

摘　　要：目的研究重组人促红细胞生成素（recombinant humanerythropoiain，rhEPO）对培养的视网膜神经节细胞（retinal ganglion cells，RGCs）存活、突起生长及生长相关蛋白43（growth associmed protein43，GAP-43）表达的影响，探讨rhEPO对培养RGCs可能的作用机制。方法分别用DMEM及含有rhEPO的DMEM对RGCs进行离体培养，观察RGCs在体外存活的时间，测量2d、4d、6dRGCs的最长突起长度；免疫细胞化学检测GAP-43蛋白表达，并测定平均灰度值。结果DMEM组离体培养RGCs于6～8d死亡，而rhEPO组RGCs能存活10～12d，存活时间较DMEM组显著延长（P〈0．01）。培养2d、4d、6d时，DMEM组最长突起长度依次为（42．90±4．71）μm、（79．74±8．49）μm、（110．02±10．79）μm，rhEPO组依次为（55．47±7．07）μm、（100．16±7．78）μm、（118．63±11．50）μm，培养2d、4d时rhEPO组与DMEM组相比差异非常显著（P〈0．01），6d时差异显著（P〈0．05）。2组细胞在培养2d时GAP-43蛋白的表达水平较高，4d时GAP-43蛋白表达到高峰。6d时GAP-43蛋白的表达水平明显降低。各时间点rhEPO组RGCsGAP-43蛋白的表达均较DMEM组有明显增高，与DMEM组相比均有非常显著差异（P〈0．01）。结论rhEPO可延长离体培养RGCs的存活时间和促进其突起的生长。上调离体培养RGCs GAP-43蛋白的表达。rhEPO促进RGCs突起生长的作用可能通过上调RGCs GAP-43蛋白的表达来实现。[眼科新进展2007；27（3）：138-192]Objective To evaluate the effect of recombinat human erythropoictin（rhEPO） on survival ,axon growth and expression of growth associated protein 43 （ CTAP-43 ） in retinal ganglion cells（ RGCs ） cultured vitro,and to investigate the possible effective mechanism of rhEPO on RGcS. Methods RGCs were cultured in DMEM and DMEM containing rhEPO in vitro respoectively,then the survival time in vitro of RGCs was oberved,the length of the longest axon of RGCs was measured on the 2nd day,the 4th day and the 6th day,the expression of GAP-43 were detected by tmmtmocytochemteal staining, the average gray scale was measured. Results RGCs died out after cultured from 6 days to 8 days in DMEM group,but RGCs could survive from 12 days to 14 days in DMEM containing rhEPO group ,the survival time of RGCs in DMEM contai- ning rhEPO group was significantly longer than that in DMEM group （P 〈 0.01 ）. After cultured on the 2nd day, the 4th day and the 6th day, the length of the longest axon of RGCs in rhEPO groupwere （55.47 ±7.07） pan, （ 100.16 ±7.78） pan and （ 118.63 ± 11.50） pan respectively, while that in DMEM group were （42.90 ± 4.71）μm, （79.74 ±8.49） μm and （110.00 ± 10.79） μm respectively,the length of the longest axon of RGCs on the 2nd day,the 4th dayin rhEPO group were longer than that in DMEM group（P 〈 0.01 ） ,and the difference on the the 6th day was very sogmofocant（P 〈 0.05 ）. The expression level of CTAP-43 in RGCs of both groups were high on the 2nd day,and reached the peak on the 4th day,trot decreased notablely on the 6th day cultured in vitro. In every time point,the average gray scale of CTAP-43 in rhEPO group were higher than that in DMEM group obviously, and there was siguificant difference （ P 〈 0.01 ）. Conclusion rhEPO can prolong the survival time,promote the axon growth and up-regulate the expression of GAP-43 of RGCs cultured in vitro stgnificantly ,which suggests that the promotive effects of rhEPO on the axon growth of RGCs may be eatab

关 键 词：重组人促红细胞生成素 视网膜神经节细胞 细胞培养 生长相关蛋白-43 

分 类 号：R774.1[医药卫生—眼科]