论文部分内容阅读
目的探讨细胞外信号调节激酶(ERK)信号转导通路在海马提取液诱导神经干细胞向神经元分化过程中的作用。方法切割12只SD大鼠右侧穹隆海马伞,14d后分别取切割侧和正常侧海马制成匀浆,离心收集上清液,蛋白定量后备用。将获取的鼠胚海马源性神经干细胞培养在24孔板中,分成3组,每组8孔,于无血清条件下,切割组加切割侧海马提取液;正常组加正常侧海马提取液;对照组不加上述海马提取液。培养14d后进行微管相关蛋白2(MAP-2)与p-ERK的免疫荧光双标检测。结果MAP-2阳性神经元数量、胞体面积和细胞周长在切割组、正常组和对照组中依次减少,3组之间均有显著性差异。MAP-2、p-ERK免疫荧光双标细胞数量在切割组、正常组、对照组中也依次下降,但双标记细胞数占MAP-2阳性神经元数的百分比却反而依次增加,两者3组之间均有显著性差异,双标细胞大多为欠成熟细胞。结论切割穹隆海马伞侧海马提取液较正常海马提取液有明显促使神经干细胞向成熟神经元分化的作用;形态学初步证实ERK信号转导通路可能与神经干细胞向神经元分化有关。
Objective To investigate the role of extracellular signal-regulated kinase (ERK) signal transduction pathway in the differentiation of neural stem cells into neurons induced by hippocampus. Methods 12 right ventricle of the hippocampus of 12 SD rats were cut. After 14 days, the hippocampus of the cut side and the normal side were made into a homogenate. The supernatant was collected by centrifugation and the protein was reserved for quantification. The obtained hippocampal-derived neural stem cells were cultured in 24-well plates and divided into 3 groups of 8 wells. Under serum-free conditions, the hippocampal slices of the hippocampus were cut and the hippocampus was harvested. The control group did not add the hippocampus extract. After cultured for 14 days, immunofluorescence double-labeled detection of MAP-2 and p-ERK was performed. Results The number of MAP-2-positive neurons, the area of the soma and the cell perimeter were decreased in the cut group, the normal group and the control group. There were significant differences among the three groups. MAP-2, p-ERK immunofluorescence double-labeled cells in the cut group, the normal group, the control group also declined, but the number of dual-labeled MAP-2 positive neurons in turn the percentage increased in turn, both 3 Significant differences between groups, double-labeled cells are mostly less mature cells. Conclusion The hippocampal extracts from the hippocampal of the diencephalons are more effective than the normal hippocampus extracts in differentiating neural stem cells from mature neurons. Morphology preliminarily confirmed that ERK signal transduction pathway may be related to differentiation of neural stem cells into neurons.