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- 2015
Rho/ROCK通路在大鼠实验性弥漫性轴索损伤中的作用
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Abstract:
摘要：目的 研究RhoA和Nogo-A在弥漫性轴索损伤(diffuse axonal injury, DAI)后的动态表达，并探讨Rho/ROCK通路抑制对DAI后脑内Nogo-A表达的调控及其在DAI中的作用。方法 用头颅瞬间旋转损伤装置制作大鼠DAI模型。观察DAI后RhoA和Nogo-A表达的时间依赖性和严重程度依赖性，并用辛伐他汀和Y27632药理性抑制RhoA/ROCK通路的活性，观察该通路在DAI后神经修复中的作用。用HE染色和PTAH染色指示DAI后脑组织病理学改变。Western blot检测RhoA和Nogo-A的表达，并检测β-APP的表达来指示轴突的损伤程度。并用免疫荧光染色指示RhoA和Nogo-A表达的空间相关性。结果 DAI后均出现明显的神经细胞坏死和轴突变性、断裂等病理改变；DAI后RhoA和Nogo-A的表达趋势一致，两者的表达量均随着DAI后时间的延长而增加，同时也随着损伤程度加重而增加，经免疫荧光双染色显示两者在脑内的表达有显著的同一性；用辛伐他汀和Y27632抑制RhoA/ROCK通路后能明显改善DAI后神经功能，并降低β-APP的表达，同时两者也能明显减少Nogo-A的表达。结论 DAI后脑内RhoA和Nogo-A的表达有高度相关性，抑制Rho/ROCK通路可改善神经功能并减轻轴突损伤。
ABSTRACT: Objective To clarify the effect of Rho/ROCK pathway inhibition on diffuse axonal injury (DAI) and its role in Nogo-A expression regulation and DAI. Methods The model of DAI by instant head rotation was produced in 52 adult rats. Twenty rats were randomly grouped according to the same severity of DAI but various duration, or the same duration but various severity. The time- and severity-dependencies of RhoA and Nogo-A expressions were investigated. HE staining and PTAH staining were used to assay the pathological changes. The expressions of RhoA and Nogo-A were detected by Western blot, and the expression of β-APP was detected for axonal injury. Immunofluorescent (IF) double staining was used to observe the locus correlation. Results There were obvious pathologic changes in the rats after DAI such as neuronal death and axonal disruption. The expressions of RhoA and Nogo-A were in the consistent trend in that the two both increased with prolonged time duration and with the increased severity of injury. IF double staining showed a significant correlation between the locuses of RhoA and Nogo-A in the brain. Both simvastatin and Y27632 improved neurorecovery through inhibiting RhoA/ROCK pathway and decreasing the expressions of β-APP and Nogo-A. Conclusion There is a significant correlation between RhoA and Nogo-A expressions, and neurorecovery can be improved by Rho/ROCK pathway inhibition
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