本文選題：α-晶狀體蛋白　切入點(diǎn)：視網(wǎng)膜神經(jīng)節(jié)細(xì)胞　出處：《第三軍醫(yī)大學(xué)》2011年碩士論文

【摘要】：目的 觀察靜脈注射α-晶體蛋白對(duì)大鼠視神經(jīng)損傷后視網(wǎng)膜神經(jīng)節(jié)細(xì)胞(retinal ganglion cells RGCs )存活和小膠質(zhì)細(xì)胞(Retinal microglia cells RMGs)活化以及對(duì)重要臟器的影響研究。 方法 1、熒光金經(jīng)雙側(cè)上丘及外側(cè)膝狀體逆行標(biāo)記RGCs,7d后制作視神經(jīng)鉗夾傷模型。視神經(jīng)鉗夾傷后經(jīng)尾靜脈分別注射1.25ml等滲生理鹽水、1×10~(-2)g/Lα-晶體蛋白、1×10~(-1)g/Lα-晶體蛋白及1g/Lα-晶體蛋白,每2天重復(fù)注射1次,共7次。傷后2周對(duì)實(shí)驗(yàn)大鼠行RGCs及RMGs計(jì)數(shù),并對(duì)肝、腎、腦、脾、肺等重要臟器進(jìn)行病理觀察。 2、在視神經(jīng)損傷模型,尾靜脈注射濃度1×10~(-1)g/L的α-晶狀體蛋白,注射方法同前,運(yùn)用熒光金逆行示蹤標(biāo)記、視網(wǎng)膜鋪片及免疫組織化學(xué)技術(shù),觀察視神經(jīng)損傷2周及4周對(duì)RGCs存活及RMGs活化的影響。 結(jié)果 1、視神經(jīng)鉗夾傷后2周RGCs數(shù)顯著下降,3個(gè)不同濃度(1×10~(-2)g/L、1×10~(-1)g/L、1g/L)α-晶體蛋白組存活的RGCs數(shù)均顯著高于生理鹽水對(duì)照組(P0.01),3個(gè)不同濃度α-晶體蛋白組活化的RMGs數(shù)均顯著低于生理鹽水對(duì)照組(P0.01)。 2、靜脈注射3個(gè)不同濃度(1×10~(-2)g/L、1×10~(-1)g/L、1g/L)α-晶體蛋白在視神經(jīng)鉗夾傷后2周對(duì)重要臟器(肝、腎、腦、脾、肺)的病理觀察未見充血、腫大、炎癥等病理改變。 3、視神經(jīng)損傷2周及4周,1×10~(-1)g/Lα-晶狀體蛋白組存活的RGCs數(shù)均顯著高于生理鹽水對(duì)照組(P0.01)。傷后2周,1×10~(-1)g/Lα-晶狀體蛋白組活化的RMGs數(shù)顯著低于生理鹽水對(duì)照組(P0.01),而傷后4周,2組間RMGs數(shù)無統(tǒng)計(jì)學(xué)差異(P0.05)。 結(jié)論 1、α-晶體蛋白靜脈注射給藥途徑對(duì)視神經(jīng)損傷2周及4周RGCs存活具有一定保護(hù)作用,表明靜脈注射是α-晶體蛋白應(yīng)用的一條有效途徑。 2、靜脈注射α-晶體蛋白能夠抑制視網(wǎng)膜小膠質(zhì)細(xì)胞反應(yīng),視神經(jīng)損傷后2周小膠質(zhì)細(xì)胞的增殖活化被抑制。 3、本研究所用靜脈注射α-晶體蛋白的濃度不會(huì)引起大鼠重要臟器大體及光鏡下的病理性改變。 4、運(yùn)用熒光金聯(lián)合免疫組化雙重標(biāo)記小膠質(zhì)細(xì)胞能夠較為準(zhǔn)確的對(duì)視神經(jīng)損傷后活化的小膠質(zhì)細(xì)胞進(jìn)行計(jì)數(shù),是研究小膠質(zhì)細(xì)胞更準(zhǔn)確有效的方法。
[Abstract]:Purpose. To observe the effects of intravenously injected 偽 -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 偽 -crystal protein 1 脳 10~(-1)g/L 偽 -crystal protein and 1 脳 10~(-1)g/L 偽 -crystal protein and 1g/L 偽 -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, 偽 -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 偽 -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 偽 -crystal protein group was significantly lower than that in saline control group (P 0.01). The number of 偽 -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) 偽 -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) 偽 -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 偽 -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 偽 -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 偽 -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, 偽 -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 偽 -crystal protein. 2. Intravenously injected 偽 -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 偽-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.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R774.1

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