蛋白激酶A抑制剂H-89及PI3-羟基激酶抑制剂对促进远端视神经损伤后视网膜神经节细胞轴突再生的影响(英文)  

Effect of protein kinase A inhibitor H-89 and PI3-kinase inhibitor on the axon regeneration of the retinal ganglion cells after distal axotomy of the optic nerve

作　　者：马宁芳[1] 李海标[2] 

机构地区：[1]广州医学院组织学与胚胎学教研室,广东省广州市510182 [2]中山医科大学组织学与胚胎学教研室,广东省广州市510080

出　　处：《中国临床康复》2006年第2期181-183,F0003,共4页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然基金资助项目(39870266)~~

摘　　要：背景:近年的研究显示中枢神经受损后在适宜的条件下如提高神经元内cAMP水平受损神经元的突起可以再生,但其确切的机制仍不清楚。目的:探讨蛋白激酶A抑制剂H-89及PI3-羟基激酶抑制剂wortman-nin对霍乱毒素促进成年金黄地鼠远端视神经受损后视网膜神经节细胞再生的影响。设计:随机对照动物实验。单位:广州医学院组胚教研室和中山医科大学组胚教研室。材料:实验于2000-01/2001-12在广州医学院和中山医科大学完成。实验动物选择健康成年雄性金黄地鼠25只。随机分为5组,手术模型组;实验对照组;霍乱毒素组;霍乱毒素+蛋白激酶A抑制剂组;霍乱毒素+PI3-羟基激酶抑制剂组,每组5只。方法:取自体坐骨神经2cm,剥去近端的神经外膜,接于视神经断端(近侧端),明胶海绵封闭窗口,坐骨神经另一端置颅骨外缝合固定于颅顶肌肉上。手术模型组视神经近侧残端对接一段自体坐骨神经。实验对照组在对照组基础上玻璃体内注射霍乱毒素溶剂Na2EDTA/NaCl;霍乱毒素组:对照组基础上玻璃体内注射霍乱毒素(1000pg/眼);霍乱毒素+蛋白激酶A抑制剂组:手术前30min玻璃体内注射3μL蛋白激酶A抑制剂H-89(60μmol/L),其余同霍乱毒素组;霍乱毒素+PI3-羟基激酶抑制剂组手术前30min玻璃体内注射3μLPI3-羟基激酶抑制剂wortmannin(1μmol/L),其余同霍乱毒素组。各组动物术后存活4周。术后每隔5天重复给药,蛋白激酶A抑制剂H89及PI3-羟基激酶抑制剂wortmannin与霍乱毒素CTx之间相隔30min注射,共4次。于取材前3d,将沾有30g/L粒蓝的明胶海绵放置在视神经断端,逆行标记再生的视网膜神经节细胞。荧光显微镜下观察,记录视网膜有轴突再生的视网膜神经节细胞数。主要观察指标:各组视网膜有轴突再生的视网膜神经节细胞数。结果:手术模型组及实验对照组有极少数视网膜神经节细胞再生(2.6±0.87,2.4±0.95),霍乱毒素BACKGROUND： Recent studies show that the neurons in central nervous system have the regenerating ability under certain suitable environment such as elevating the c-AMP level of the neuron. But the mechanism is unclear. OBJECTIVE： To investigate the effect of protein kinase A inhibitor H-89 and PI3-kinase inhibitor, wortmannin, on Cholera Toxin （CTx） in promoting the axon regeneration of retinal ganglion cells （RGCs） after distal axotomy of the optic nerve in adult hamstem. DESIGN： A randomized and controlled animal experiment SETTING： Department of Histology and Embryology of Guangzhou Medical College and Sun Yat-sen University of Medical Sciences MATERIALS： This experiment was conducted in the Guangzhou Medical College and Sun Yat-sen University of Medical Sciences between January 2000 and December 2001. Totally 25healthy adult male hamsters were chosen and randomly divided into 5 groups： model group; experimental control group; CTx group; CTx ＋protein kinase A inhibitor group; CTx ＋ PI3-K inhibitor group, with 5 animals in each group. METHODS： A 2 cm segment of autologns sciatic nerve was removed and desheathed. The proximal end of the sciatic nerve was connected to the proximal stump of the optic nerve （ON）. The wound was enveloped with gelatin sponge. The remaining portion of sciatic nerve was placed on the top of the skull. Model group： a segment of autologns sciatic nerve was connected to the ON proximal stump. Experimental control group： Received an injection of Na2EDTA/ aCL solution introvitreally based on the treatment in control group. CTx group： CTx （1000 pg/eye）was injected introvitreally on the basis of the treatment in the control group. CTx＋ protein kinase A inhibitor group ： 3μ protein kinase A inhibitor H89 （60 μmol/L）was injected introvitreally 30 minutes before operation, the other treatment was like that of CTx group. ⑤CTx＋PI3-K inhibitor group ： 3 o,L PI3-K inhibitor wortmannin （1μmol/L） was injected introvitreally 30 minutes before

关 键 词：视网膜神经节细胞 霍乱毒素 轴突 

分 类 号：R774.1[医药卫生—眼科]