雪旺细胞源性神经营养活性物质对培养视网膜节细胞正常及缺气损伤环境中存活及生长的作用  

Effects of Schwann Cell-derived Neurotrophic Acti - vity on Cultured Retinal Ganglion Cells Survive and Grow in Normal and Gas-deprived Environment

作　　者：黄蔚[1] 王琳[1] 惠延年[1] 马吉献[1] 

机构地区：[1]第四军医大学西京医院全军眼科研究所

出　　处：《眼科学报》2002年第4期235-239,共5页Eye Science

基　　金：国家自然科学基金:NO.39900076

摘　　要：目的:探讨雪旺细胞(Schwann cells,SC)源营养神经活性物质(SC derived neurotrophic activity,SCNA)对体外培养视网膜节细胞正常及缺气损伤环境中存活及生长相关蛋白(GAP)_(43)表达的作用。方法:培养乳鼠SC,制备不同蛋白浓度的SCNA,加入原代培养的视网膜细胞中,MTT法检测SCNA活性培养荧光金逆行标记的新生3d的SD乳鼠视网膜细胞,接种于24孔板中。培养第2d时SCNA作用组培养液中加入300mg/l。SCNA,对照组不加;培养第5d缺气损伤组在培养液表面加入液体石蜡造成缺气损伤。12h后去除液体石蜡,观察细胞形态和计数荧光金逆行标记的视网膜节细胞(retinal ganglion cell,RGC)。Western Blot法分析SCNA对视网膜细胞GAP_(43)表达的影响。结果:SCNA能促进培养视网膜细胞的存活,并有蛋白浓度依赖关系。加SCNA后,视网膜细胞生长明显旺盛,悬浮的死细胞较少,RGC数量明显多于视网膜细胞单纯培养组(F=62.89,P<0.01)。缺气损伤组视网膜细胞出现肿胀改变,加有SCNA缺气损伤组细胞形态大致正常,RGC数与对照组相比有显著性差异(F=49.27,P<0.01)。GAP_(43)的表达在视网膜细胞正常及缺气损伤SCNA作用组上调。结论:SCNA对培养RGC具有明显营养作用,并上调GAP_(43)的表达。在培养液中加入SCNA,可以减轻“缺气”造成的损伤,提高体外培?Purpose: To investigate the effects of Schwann cell-derived neurotrophic activity (SCNA) on cultured retinal ganglion cells (RGC) survive and the expression of growth associated protein 43 (GAP43) in normal and gas-deprived environment. Methods: Schwann cells of new born SD rat were cultured, and remaded it into solutions containing different concentrations of SCNA. Then these solutions were added to the primarily cultured retinal cells, the actvity of SCNA was determined by MTT method. The fluro-gold retrolabelled retina cells of 3d new born rats were cultured and plated on 24 well culture plate. At the 2rd day of culture, 300mg/L SCNA were added into the medium of SCNA group. At the 5th day, liquid paraffin was added on the surface ofmedium of air deprived groups to lead an air deprived injury. Cultured for 12h, the liquid paraffin was discarded off, and then we observed the morphology and cunted the fluro-gold labeled RGC The effects of SCNA on the expression of GA?43 in cultured retinal cells were identified by Western Blot. Result: SCNA could promote the survival of RGC in a protein concentration-dependant manner. Added SCNA to the medium, the retinal cells grown much better and had less floating dead cells. The number of RGC was also significantly more than the control ( F = 62. 89, P < 0. 01). The cells of air deprived group had showed swelling change. But the cells of group with SCNA are less affected and the number of RGCs is significantly different from the contrl ( F = 49. 27, P < 0. 01). The expression of GA?43 in normal and air deprived groups with SCNA were up-regulated. Conclusion: SCNA has significantly neurotrophic effect on cultured RGC and increases the expression of GAP4a. Added SCNA to the medum could reduce the injury of air deprived and promote RGC survival rate in this kind of injury environment. Eye Science 2002; 18: 235 - 239.

关 键 词：神经营养因子 雪旺细胞 视网膜节细胞 动物实验 RGC 

分 类 号：R770.2[医药卫生—眼科]