α-晶体蛋白保护视网膜神经节细胞存活及抑制小胶质细胞活化的在体研究

发布时间：2018-04-01 07:34

本文选题：α-晶状体蛋白　切入点：视网膜神经节细胞　出处：《第三军医大学》2011年硕士论文

【摘要】：目的观察静脉注射α-晶体蛋白对大鼠视神经损伤后视网膜神经节细胞(retinal ganglion cells RGCs )存活和小胶质细胞(Retinal microglia cells RMGs)活化以及对重要脏器的影响研究。方法 1、荧光金经双侧上丘及外侧膝状体逆行标记RGCs,7d后制作视神经钳夹伤模型。视神经钳夹伤后经尾静脉分别注射1.25ml等渗生理盐水、1×10~(-2)g/Lα-晶体蛋白、1×10~(-1)g/Lα-晶体蛋白及1g/Lα-晶体蛋白,每2天重复注射1次,共7次。伤后2周对实验大鼠行RGCs及RMGs计数,并对肝、肾、脑、脾、肺等重要脏器进行病理观察。 2、在视神经损伤模型,尾静脉注射浓度1×10~(-1)g/L的α-晶状体蛋白,注射方法同前,运用荧光金逆行示踪标记、视网膜铺片及免疫组织化学技术,观察视神经损伤2周及4周对RGCs存活及RMGs活化的影响。结果 1、视神经钳夹伤后2周RGCs数显著下降,3个不同浓度(1×10~(-2)g/L、1×10~(-1)g/L、1g/L)α-晶体蛋白组存活的RGCs数均显著高于生理盐水对照组(P0.01),3个不同浓度α-晶体蛋白组活化的RMGs数均显著低于生理盐水对照组(P0.01)。 2、静脉注射3个不同浓度(1×10~(-2)g/L、1×10~(-1)g/L、1g/L)α-晶体蛋白在视神经钳夹伤后2周对重要脏器(肝、肾、脑、脾、肺)的病理观察未见充血、肿大、炎症等病理改变。 3、视神经损伤2周及4周,1×10~(-1)g/Lα-晶状体蛋白组存活的RGCs数均显著高于生理盐水对照组(P0.01)。伤后2周,1×10~(-1)g/Lα-晶状体蛋白组活化的RMGs数显著低于生理盐水对照组(P0.01),而伤后4周,2组间RMGs数无统计学差异(P0.05)。结论 1、α-晶体蛋白静脉注射给药途径对视神经损伤2周及4周RGCs存活具有一定保护作用,表明静脉注射是α-晶体蛋白应用的一条有效途径。 2、静脉注射α-晶体蛋白能够抑制视网膜小胶质细胞反应,视神经损伤后2周小胶质细胞的增殖活化被抑制。 3、本研究所用静脉注射α-晶体蛋白的浓度不会引起大鼠重要脏器大体及光镜下的病理性改变。 4、运用荧光金联合免疫组化双重标记小胶质细胞能够较为准确的对视神经损伤后活化的小胶质细胞进行计数,是研究小胶质细胞更准确有效的方法。
[Abstract]:Purpose. To observe the effects of intravenously injected 伪 -crystal protein on the survival of retinal ganglion cells RGCs and the activation of microglial microglia cells RMGs after optic nerve injury in rats. Method. 1. The model of optic nerve clamp injury was made by retrograde labeling of RGCs through bilateral superior colliculus and lateral geniculate body for 7 days. 1 脳 10~(-2)g/L 伪 -crystal protein 1 脳 10~(-1)g/L 伪 -crystal protein and 1 脳 10~(-1)g/L 伪 -crystal protein and 1g/L 伪 -crystal protein were injected into the caudal vein after optic nerve clamp injury. RGCs and RMGs were counted 2 weeks after injury, and liver, kidney, brain, spleen, lung and other important organs were observed. 2. In the optic nerve injury model, 伪 -crystallin with 1 脳 10~(-1)g/L concentration was injected into the caudal vein with the same method as before. Fluorescent gold retrograde tracing, retinal smear and immunohistochemistry were used. The effects of optic nerve injury on RGCs survival and RMGs activation were observed at 2 and 4 weeks after injury. Results. 1. The number of RGCs decreased significantly 2 weeks after optic nerve clamp injury. The RGCs number of 伪 -crystal protein group was significantly higher than that of normal saline control group (P 0.01). The number of activated RMGs in three different concentrations of 伪 -crystal protein group was significantly lower than that in saline control group (P 0.01). The number of 伪 -crystal protein activated in three groups was significantly lower than that in normal saline control group (1 脳 10 ~ (-1) g 路L ~ (-1) 路L ~ (-1)) ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 伪 -crystal protein. 2. Intravenously injected 3 different concentrations of 1 脳 10 ~ (-1) ~ (-2) g / L ~ (1 脳 10 ~ (-1)) ~ (-1) g 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 伪 -crystal protein did not show the pathological changes of important organs (liver, kidney, brain, spleen, lung) in 2 weeks after optic nerve clamp injury, such as congestion, swelling, inflammation and so on. 3. The number of RGCs surviving in the 1 脳 10~(-1)g/L 伪 -crystallin group was significantly higher than that in the saline control group at 2 and 4 weeks after injury, and the number of activated RMGs in the 1 脳 10~(-1)g/L 伪 -crystallin group was significantly lower than that in the saline control group at 2 and 4 weeks after injury, while the number of activated RMGs in the 1 脳 10~(-1)g/L 伪 -crystallin group was significantly lower than that in the saline control group at 4 weeks after injury. There was no significant difference in the number of RMGs between groups (P 0.05). Conclusion. 1, 伪 -crystal protein was injected intravenously to protect the survival of RGCs for 2 and 4 weeks after optic nerve injury, indicating that intravenous injection is an effective way to use 伪 -crystal protein. 2. Intravenously injected 伪 -crystal protein could inhibit retinal microglia reaction, and the proliferation and activation of microglia were inhibited 2 weeks after optic nerve injury. 3. The concentration of 伪-crystal protein by intravenous injection did not cause pathological changes in important organs of rats under light microscope. 4. Fluorescent gold combined with double labeling of microglia with immunohistochemistry can accurately count the activated microglia after optic nerve injury, which is a more accurate and effective method to study microglia.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R774.1

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