重组人促红细胞生成素对大鼠视网膜神经节细胞的影响

Effect of recombinant human erythropoietin on rat retinal ganglion cells

目的:观察重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对体外培养的大鼠视网膜神经节细胞(retinalganglioncells,RGCs)存活及其轴突生长的影响,检测作为神经再生标志的生长相关蛋白43(growth associated protein 43,GAP-43)的表达情况,并探讨rhEPO对RGCs可能的作用机制。方法:RGCs分为实验组(rhEPO组)和对照组(DMEM组)行体外培养,倒置显微镜下观察细胞和轴突生长情况,培养72h测量细胞最长突起长度进行比较,行GAP-43的Western-blot检测,图像分析系统对两组标本进行灰度值测量。结果:培养72h的RGCs倒置显微镜下观察,形成典型的细胞突起,rhEPO组的细胞比对照组胞体更大,突起更长(P<0.05)。培养72h的RGCs行GAP-43Western-blot检测,DMEM组GAP-43呈阳性表达,rhEPO组呈强阳性表达,两组差异有统计学意义(P<0.05)。结论:rhEPO能够促进体外培养的RGCs轴突生长,能够上调体外培养的RGCs的GAP-43蛋白表达水平,可能是其能够促进RGCs轴突生长的原因。

AIM： To observe the influence of recombinant human erythropoietin （rhEPO） on cultured retinal ganglion cells （RGCs） survival and axonal regeneration of rats, detect the expression of growth associated protein 43 （GAP-43）, and to investigate the possible effective mechanism.METHODS：RGCs were cultured in DMEM （control group） or DMEM containing rhEPO （rhEPO group） for 72 hours. Cell morphology and axonal growth were observed under phase-contrast microscope. The length of the longest processes of RGCS were measured and compared between two groups. The GAP-43 expression was detected by Western blot and the gray value scales of GAP-43 were measured by imaging analysis system. All results were compared using t test.RESULTS：The RGCs were observed to extend processes under microscope after culturing for 72 hours. The cells in rhEPO group were bigger and the processes were longer compared to the control group （P0.05）. The expression of GAP-43 in vitro was detected by Western blot.The expression of GAP-43 in rhEPO group was higher than that in control group （P0.05）.CONCLUSION： rhEPO can promote the axonal growth of cultured RGCs in vitro. rhEPO can also increase the expression of GAP-43 in cultured RGCs, which may result in the promotion of rhEPO on the axonal growth of RGCs.