siRNA靶向沉默Vimentin抑制大鼠急性脊髓损伤胶质瘢痕形成的研究

Spinal cord injury is one of serious central nervous system diseases, which an increasing number of victims were suffered from all over the world. Unfortunately, efficient methods had not been found for dealing with it. People found that Vimentin expression would elevate after spinal cord injury, which always have expressed during development period. In order to proof the effects of Vimentin in glial scar after acute spinal cord injury, the expression of Vimentin will be silenced by RNA interference. The in vitro and in vivo mechanism of Vimentin after acute spinal cord injury would be searched. Furthermore, the association between reactive astrogliosis, neuron microenvironment, glial scar formation and Vimentin gene will be studied further. We will verify the effects of Vimentin in acute spinal cord injury rats’ models. The purpose of this study is to scientifically analysis the effects of Vimentin from molecular, cellular and animal level. The results of the study should provide both the experimental evidence and theoretical witness for RNA interference Vimentin dealing with spinal cord injury. What is more, it would be verified to be a efficient method to cure spinal cord injury.

脊髓损伤（SCI）是中枢神经系统严重创伤，发生率在全球呈上升趋势，目前尚无确切有效治疗方法。研究发现胶质瘢痕形成过程中表达于发育期的中间丝波形蛋白(Vimentin)重新表达，为了验证Vimentin基因在胶质瘢痕形成及急性脊髓损伤过程中的作用，本研究项目利用siRNA技术，选择性沉默Vimentin基因表达，验证其在体内、外的作用机制。深入研究Vimentin在急性脊髓损伤后，对反应性星形胶质细胞增生、神经元细胞生长微环境以及胶质瘢痕形成的影响。并在急性SCI动物模型当中进一步验证该基因的作用对胶质瘢痕形成及脊髓损伤修复的影响。从动物整体、细胞和分子水平系统分析Vimentin在胶质瘢痕形成及急性脊髓损伤过程当中的作用机制。可进一步阐明脊髓损伤后胶质瘢痕形成的机制，并且研究项目也为RNA干扰Vimentin治疗SCI的临床应用提供了实验基础和理论依据，可能开拓一种治疗SCI的新途径。

脊髓损伤（SCI）是中枢神经系统严重创伤，发生率在全球呈上升趋势，目前尚无确切有效治疗方法。反应性星形胶质细胞瘢痕一直被认为是脊髓损伤过程当中的不利因素，主要由于在中枢神经系统创伤当中瘢痕形成物理屏障抑制轴突跨越损伤部位。研究发现胶质瘢痕形成过程中表达于发育期的中间丝波形蛋白(Vimentin)重新表达，为了验证Vimentin基因在胶质瘢痕形成及急性脊髓损伤过程中的作用，本研究项目利用siRNA技术，选择性沉默Vimentin基因表达，验证其在体内、外的作用机制。 . 研究主要包括：设计合成Vim干扰siRNA质粒，进行鉴定，并研究其功能，其中siRNA-Vim-1073相对沉默效果最好；siRNA靶向沉默Vim对大鼠星形胶质细胞活性、生长周期、功能基因表达及反应性星形胶质细胞瘢痕的影响；siRNA靶向沉默Vim对大鼠脊髓神经元细胞活性及细胞生长微环境的影响；建立SD 大鼠急性脊髓损伤模型，评价siRNA 靶向沉默Vim 对SCI 大鼠胶质瘢痕形成、功能基因表达、轴突再生情况的影响，并从行为学、免疫组织化学、神经电生理等角度研究siRNA 靶向沉默Vim 的作用效果。. 研究结果表明：siRNA作用于Vimentin能够下调星形胶质细胞表达GFAP、CSPG，从而抑制反应性星形胶质细胞增生；脊髓损伤早期局部注射siRNA-Vimentin可以使脊髓损伤部位GFAP、CSPG和Vimentin表达下调；GFAP、CSPG和Vimentin协同发挥作用，抑制脊髓损伤后胶质瘢痕的形成，为脊髓损伤后轴突再生及神经功能恢复提供良好的微环境。动物实验的结果也进一步证实siRNA-Vimentin对促进脊髓损伤大鼠运动功能及神经电生理功能的恢复有明显的促进作用。并且研究项目为RNA干扰Vimentin治疗SCI的临床应用提供了实验基础和理论依据，可能开拓一种治疗SCI的新途径。