晶状体损伤促进视神经损伤后轴突再生机制的研究

发布时间：2018-07-06 19:06

本文选题：生长相关蛋白43 + 晶状体损伤　；参考：《郑州大学》2011年硕士论文

【摘要】：视神经损伤(optic nerve injury)是眼科常见的一种临床眼外伤类型及致盲原因,常见于各种面部、颅脑及眼眶外伤,如压迫、牵拉、切断等原因均能导致视神经严重受损,可能造成患者视功能不可逆性的丧失。视神经损伤后大量视网膜神经节细胞(retinal ganglion cells, RGCs)的坏死或凋亡,是视力丧失的主要原因。减缓或抑制RGCs的死亡及促进其轴突的再生,是有效治疗视神经损伤和促进视功能恢复的基础。因此视神经损伤再生已成为当前国内外研究的热点。传统认为成年哺乳动物的中枢神经系统损伤后不能再生,损伤后的神经元难以形成新的突触连接。20世纪80年代,Hall等人用一段自体坐骨神经移植到视神经损伤区,并成功地诱导激发了视神经损伤后轴突的再生,中枢神经损伤后不能再生的观念发生了改变。近年来Leon和Fischer等研究发现,单纯晶状体损伤能大大增加RGCs的存活率,并能促进视神经损伤后的轴突再生。因而晶状体损伤促进视神经损伤再生的机制研究,受到国内外学者的广泛关注。研究认为激活的巨噬细胞在外周神经损伤后,具有吞噬、合成和分泌多种活性因子等作用,形成有利于神经再生的内环境,从而促进外周神经的再生,但巨噬细胞在中枢神经损伤再生中是否也有这种作用,晶状体损伤能否激活巨噬细胞在视神经损伤再生中发挥作用研究很少。本实验采用大鼠在体实验研究的方法进行进一步探讨和研究。目的研究晶状体损伤促进视神经损伤后视网膜节细胞轴突再生的作用,并探讨活化的巨噬细胞所起作用,为视神经损伤后治疗方面提供新的思路。方法选用普通级成年健康Wistar大鼠50只作为实验动物,随机分为4组：正常对照组5只不做任何处理；单纯视神经横断伤(NC)组15只；联合晶状体损伤(NC+LP)组15只；联合玻璃体腔注射酵母多糖(NC+ZI)组15只。分别在术后1周、2周、3周后处死大鼠5只,通过免疫组化方法应用ED-1单克隆抗体,标记视网膜内激活的巨噬细胞,及免疫荧光方法标记视网膜生长相关蛋白-43(growth associated protein 43, GAP-43)的表达,观察并记录不同时间点阳性荧光着色面积。结果正常对照组及NC组,在视网膜的各层中未见ED-1阳性的巨噬细胞,NC+LP组及NC+ZI组,在视网膜的各层中可见ED-1阳性巨噬细胞。视网膜中GAP-43表达：正常对照组仅见少量GAP-43荧光素着色,主要集中在内丛状层,视网膜节细胞层未见着色。NC组术后1周时视网膜节细胞层可见少量GAP-43荧光素染色,随时间延长逐渐减少,在2周时基本消失。NC+LP组及NC+ZI组术后视网膜节细胞层可见大量GAP-43荧光素着色,术后前2周GAP-43着色面积较明显,至3周时仍有着色。 NC组与正常组视网膜GAP-43阳性细胞着色面积相比,在1周时有显著统计学差异(P0.01)；在2、3周时无统计学差异(P0.05)。 NC组分别与NC+LP组及NC+ZI组在不同时间点视网膜GAP-43阳性着色面积均有显著统计学差异(P0.01)；均在伤后早期GAP-43表达最多,以后其表达量逐渐下降。 NC+LP组与正常组相比,视网膜GAP-43阳性细胞着色面积均有显著统计学差异(P0.01)；与NC+ZI组相比,1周时无统计学差异(P0.05),2、3周时均有统计学差异(P0.05)。 NC+ZI组与正常组视网膜GAP-43阳性细胞着色面积不同时间点均有显著统计学差异(P0.01)： NC+LP组(及NC+ZI组)在各时间点间相比较视网膜GAP-43阳性细胞着色而积均有显著统计学差异(P0.01)。结论 1.单纯视神经损伤后,早期仅见少量视网膜反应性GAP-43的表达增多。 2.晶状体损伤和玻璃体内注射酵母多糖可以激活巨噬细胞,使视神经损伤后视网膜中GAP-43的表达明显增多,对视神经的轴突再生具有促进作用,在损伤后前两周时作用较明显,可持续至第3周。
[Abstract]:optic nerve injury ( optic nerve injury ) is a common type and cause of blindness in ophthalmology . It is common in various facial , head and orbital trauma , such as compression , traction , cutting , etc .

In recent years , it has been found that the activation of macrophages can significantly increase the survival rate of RGCs and promote axonal regeneration after optic nerve injury . In recent years , it has been found that the activation of macrophages can significantly increase the survival rate of optic nerve and promote axonal regeneration after optic nerve injury .

Purpose

To study the effect of lens injury on retinal ganglion cell axonal regeneration after optic nerve injury , and to explore the role of activated macrophages in the treatment of optic nerve injury .

method

50 adult healthy Wistar rats were randomly divided into four groups : normal control group and normal control group .
15 cases of simple optic nerve transection injury ( NC ) group ;
15 patients with combined lens injury ( NC + LP ) ;
Fifteen rats were sacrificed at 1 week , 2 weeks and 3 weeks after surgery . The expression of the retinal growth related protein 43 ( GAP - 43 ) was marked by immunohistochemical method , and the stained area of positive fluorescence was observed and recorded .

Results

In normal control group and NC group , ED - 1 positive macrophages , NC + LP group and NC + ZI group were not seen in each layer of retina , and ED - 1 positive macrophages were seen in each layer of retina .

GAP - 43 expression in the retina was observed : only a small amount of GAP - 43 fluorescein staining was observed in the normal control group . The staining of GAP - 43 fluorescein was observed in the retinal ganglion cell layer at 1 week after operation .

Compared with the stained area of GAP - 43 positive cells in NC group , there was significant difference between NC group and normal group retinal GAP - 43 positive cells ( P0.01 ) .
There was no statistical difference between 2 and 3 weeks ( P0.05 ) .

There was significant difference between NC group and NC + LP group and NC + ZI group at different time points ( P0.01 ) .
The expression of GAP - 43 at the early stage of injury was the most , and the expression of GAP - 43 decreased gradually in the future .

Compared with the normal group , the staining area of GAP - 43 positive cells was significantly different between NC + LP group and normal group ( P0.01 ) .
There was no statistical difference ( P0.05 ) at 1 week compared with NC + ZI group ( P0.05 ) .

The staining area of GAP - 43 positive cells in NC + ZI group was significantly different from that in normal group ( P0.01 ) .

The GAP - 43 positive cells were positive in NC + LP group ( NC + ZI group ) at each time point .

There was significant difference between color and color ( P0.01 ) .

Conclusion

1 . After optic nerve injury , only a small amount of retinal reactive GAP - 43 was seen in the early stage .

2 . Lens damage and intravitreous injection of yeast polysaccharide can activate macrophages , and the expression of GAP - 43 in the retina after optic nerve injury is obviously increased , and the axonal regeneration of optic nerve can be promoted , and the effect is obvious two weeks after injury , which can last until the third week .
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R779.1

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