a晶体蛋白调节星形胶质细胞活化在促视神经轴突再生的作用及机制研究

The optic nerve damage and repair is an eye foundation research difficulty and hot. The formation of glial scar by astrocytes activation is the main reason that optic nerve axons can not regenerate after injury. Preliminary studies have shown that α-crystallin can promote injuryed renewable axons across the damage area, also observed α-crystallin inhibits GFAP expression of astrocyte, but the relationship between the two facts is not yet clear. BMPs / Smads is a key signaling pathways that regulate the activation of astrocytes. This research intends to know whether α-crystallin play a role to promote axonal regeneration, through the impact of BMPs / Smads signaling pathway, resulting in inhibiting the activation of astrocytes to reduce glial scar and nerve inhibitor and inflammatory cytokine release, by using immunofluosrescence confocal laser, the recombinant adenovirus transfection method、RT-PCR、Western blot analysis in cultured rat optic nerve astrocytes and optic nerve incomplete injury model. This research further reveal regeneration way of α-crystallin facilitating the optic nerve axonal and possible molecular mechanism. This research is the continuation and depth of the preliminary work , contribute to a comprehensive exposition of the mechanism of the α-crystallin to promote optic nerve regeneration, reveal new targets of α-crystallin and provide a theoretical basis for the application of the α-crystallin.

视神经损伤后星形胶质细胞活化形成的胶质疤痕是视神经轴突不能再生的主要原因之一。研究证明：α晶体蛋白能促进视神经损伤后再生轴突穿越损伤区，但机理不清。前期发现：a晶体蛋白可抑制星形胶质细胞增殖及GFAP表达。鉴于BMPs/Smads是调节星形胶质细胞活化的关键信号通路，提示α晶体蛋白可能调节星形胶质细胞而促神经再生。本课题拟通过大鼠视神经星形胶质细胞培养和视神经不全损伤模型，采用免疫荧光激光共聚焦、over-expression 和siRNA等方法探讨α晶体蛋白是否可以通过对BMPs/Smads信号通路的影响，调节星形胶质细胞的活化，减少胶质疤痕及神经抑制物和炎性因子的释放而发挥促进轴突再生的作用，进一步揭示α晶体蛋白促进视神经轴突再生途径和可能的分子机制。本项目是前期工作的继续和深入，有助于全面阐述α晶体蛋白促视神经再生的作用机制，揭示α晶体蛋白新的作用靶点，为α晶体蛋白应用提供依据。

视神经损伤后星形胶质细胞活化形成的胶质疤痕是视神经轴突不能再生的主要原因之一。研究证明：α晶体蛋白能促进视神经损伤后再生轴突穿越损伤区，但机理不清。前期发现：a 晶体蛋白可抑制星形胶质细胞增殖及 GFAP 表达。鉴于 BMPs/Smads 是调节星形胶质细胞活化的关键信号通路，提示α-晶体蛋白可能调可节星形胶质细胞而促神经再生。本课题通过对大鼠视神经星形胶质细胞培养和视神经不全损伤模型，体外应用脂多糖、划痕和糖氧剥夺刺激视神经星形胶质细胞，采用免疫荧光激光共聚焦、ELISA和 siRNA 等方法证实α晶体蛋白可能通过对 BMPs/Smads 信号通路的影响，调节星形胶质细胞的活化，减少胶质疤痕及神经抑制物和炎性因子的释放而发挥促进轴突再生的作用，进一步揭示α晶体蛋白促进视神经轴突再生途径和可能的分子机制。本文阐述α-晶体蛋白促视神经再生的可能作用机制，揭示α晶体蛋白新的作用靶点，为α晶体蛋白的临床应用提供了一定理论依据。