体外切割损伤脊髓神经元对其Fos、ChAT及GAD表达的影响

目的:研究切割损伤对体外培养脊髓神经元Fos、胆碱乙酰转移酶(ChAT)及谷氨酸脱羧酶(GAD)表达的影响。方法:脊髓神经元纯化培养后分别计数碘化丙啶(PI)阳性细胞数和微管相关蛋白2(MAP2)阳性细胞数,决定脊髓神经元纯度。根据本实验室已建立的损伤模型,对原代纯化培养脊髓神经元进行切割损伤。通过免疫组化及WesternBlot方法比较神经元损伤前后Fos、ChAT和GAD表达的变化情况。结果:脊髓神经元纯化培养后纯度达到92%。免疫组化及WesternBlot显示,切割损伤组与正常组相比,Fos、GAD表达上调,而ChAT表达降低。结论:Fos、GAD和ChAT可能参与脊髓神经元急性损伤及随后的修复过程,其生物学意义有待进一步探讨。

Effects of incision in vitro on expression of Fos.ChAT and GAD in spinal cord neuron cells

Objective:To explore effects of incision injury to cultured spinal cord neuron cells on expressions of Fos,ChAT(choline acetyltransferase)and GAD(glutamate decarboxylase).Methods:The purity of the spinal cord neuron culture was determined according to account numbers of microtubule-associated protein-2(MAP2)-positive cells in propidium iodide(PI)-stained cells.Based on our established incision injury model,the purified spinal cord neuron cells were incised and expression levels of Fos,ChAT and GAD in the incised neuronal cells were compared with those in uninjured cells through immunohistochemical staining and Western Blot.Results:The spinal cord neuron cultures were comprised of 92% neurons.The expression levels of Fos and GAD were upregulated but ChAT downregulated through immunohistochemical staining and Western Blot.Conclusion:Fos、GAD and ChAT may be involved in process of acute injury and subsequent recovery of spinal cord neuron cells,and its biological significances should be further explored.