Colivelin对视神经保护作用的初步研究

发布时间：2018-06-27 10:25

本文选题：Colivelin + 外伤性视神经病变　；参考：《眼科新进展》2012年04期

【摘要】：目的初步探讨Colivelin对大鼠外伤性视神经损伤后的神经保护作用和机理,及其保护效果是否具有剂量依赖性。方法 40只健康2月龄Wistar大鼠,随机分为模型组、安慰剂组及Colivelin低剂量组(10-6 μmol·L-1 Colivelin)、Colivelin中剂量组(10-4μmol·L-1Colivelin)、Colivelin高剂量组(10-2 μmol·L-1 Colivelin),每组8只。40只大鼠均做单侧视神经损伤模型,模型组另一侧未做任何处理的8眼为空白对照组。应用无创血管夹建立大鼠视神经夹伤模型,造模后30 min Colivelin治疗组玻璃体内分别注射不同浓度Colivelin 5 μL,安慰剂组注射5μLPBS缓冲液,模型组和空白对照组不给予任何治疗。注药后7d,通过视网膜切片技术,进行HE染色观察视网膜神经节细胞(retinal ganglion cell,RGC)的形态及数目,TUNEL染色法检测RGC的凋亡以及免疫组织化学法检测视网膜中caspase-3的表达。结果 HE染色可见空白对照组视网膜各层细胞排列整齐密集,模型组、安慰剂组、Colivelin治疗组均可见部分RGC核固缩,但随着Colivelin注射浓度的增加,RGC发生核固缩的数量减少。RGC计数:空白对照组每400倍光镜视野下RGC数量为25.750±1.264,模型组为8.236±1.239,安慰剂组为8.514±1.222,Colivelin低剂量组为14.500±1.021,Colivelin中剂量组为16.250±1.319,Colivelin高剂量组为18.097±1.323,除模型组与安慰剂组之间差异无统计学意义(P0.05)外,其他各组之间差异均有统计学意义(均为P0.05)。TUNEL染色:模型组和安慰剂组注药后7 d TUNEL染色可见大量凋亡细胞,Colivelin治疗组凋亡细胞数量随Colivelin注射浓度的增加依次递减。RGC细胞凋亡率:模型组为(60.928±2.961)%,安慰剂组为(61.446±2.755)%,Colivelin低剂量组为(58.432±2.835)%,Colivelin中剂量组为(51.948±2.802)%,Colivelin高剂量组为(47.656±2.331)%。Caspase-3表达:模型组、安慰剂组及Colivelin低剂量组、中剂量组、高剂量组均可见caspase-3表达。平均光密度值显示,模型组和安慰剂组相比caspase-3表达无明显差异(分别为0.482±0.012和0.486±0.012),均高于Colivelin治疗组(均为P0.05),并且Colivelin低剂量组、中剂量组、高剂量组组间差异亦均有统计学意义(分别为0.411±0.017、0.326±0.018、0.234±0.016;均为P0.05)。结论 Colivelin能有效增加视神经损伤后RGC的数量,抑制其凋亡,减少caspase-3表达,且这一作用呈剂量依赖性。
[Abstract]:Objective to investigate the neuroprotective effect and mechanism of Colivelin on traumatic optic nerve injury in rats and whether the protective effect is dose-dependent. Methods Forty healthy 2-month-old Wistar rats were randomly divided into three groups: model group, placebo group and Colivelin low dose group (10-6 渭 mol L-1 Colivelin). The model of unilateral optic nerve injury was established in 10 ~ 4 渭 mol L ~ (-1) Colivelin high-dose group (10 ~ (-2) 渭 mol ~ (-1) Colivelin). 8 eyes in the model group without any treatment on the other side were blank control group. A rat model of optic nerve injury was established by using a non-invasive vascular clamp. 30 min after the establishment of the model, different concentrations of Colivelin 5 渭 L were injected into the vitreous of the model group and 5 渭 L PBS buffer solution was injected into the placebo group. The model group and the blank control group were not given any treatment. At 7 days after injection, the morphology and number of retinal ganglion cells (retinal ganglion cells) were observed by HE staining. The apoptosis of retinal ganglion cells was detected by Tunel staining and the expression of caspase-3 in retina was detected by immunohistochemistry. Results HE staining showed that all layers of retinal cells in blank control group were arranged neatly and densely, and partial RGC nuclear contractions were observed in both the model group and the placebo group. However, with the increase of Colivelin injection concentration, the number of nuclear contractions in RGCs decreased. RGC count: the number of RGCs in the blank control group was 25.750 卤1.264 per 400-fold light microscope, in the model group was 8.236 卤1.239, and in the placebo group was 8.514 卤1.222um Colivelin, 14.500 卤1.021Colivelin in the middle dose group and 16.250 卤1.319m Colivelin high dose group. There was no significant difference between the model group and the placebo group (P0.05). There were significant differences between the other groups (P0.05). Tunel staining: Tunel staining showed that the number of apoptotic cells decreased with the increase of Colivelin injection concentration in model group and placebo group 7 days after injection. RGC showed that the number of apoptotic cells in Colivelin treatment group decreased with the increase of Colivelin injection concentration. The rate of apoptosis in the model group was (60.928 卤2.961) and in the placebo group was (61.446 卤2.755). The expression of Caspase-3 in the low dose group was (58.432 卤2.835) vs (51.948 卤2.802) in the middle dose group and (47.656 卤2.331) in the high dose group. Caspase-3 expression was observed in placebo group, Colivelin low dose group, middle dose group and high dose group. The mean optical density showed that there was no significant difference in caspase-3 expression between model group and placebo group (0.482 卤0.012 and 0.486 卤0.012, respectively), which was higher than that of Colivelin treatment group (P0.05). The difference of high dose group was also statistically significant (0.411 卤0.017) 0.326 卤0.018 ~ 0.234 卤0.016; all were P0.05). Conclusion Colivelin can effectively increase the number of RGCs after optic nerve injury, inhibit their apoptosis and reduce the expression of caspase-3 in a dose-dependent manner.
【作者单位】：广西医科大学第一附属医院眼科;
【分类号】：R774.6

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