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大鼠胚胎小脑提取液对培养脊髓神经细胞的作用 |
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吕捷 吕永利
摘要 目的：探讨不同胎龄大鼠小脑提取液(CE)对体外培养脊髓神经细胞的影响。方法：制备不同蛋白浓度的14，16，18，20 d胎鼠(E14、E16、E18、E20)CE，加入原代培养的E18胎鼠脊髓神经细胞中，24 h后以微量快速自动比色法检测细胞活性。结果：实验组各胎龄CE均能促进培养脊髓神经元的存活，并有蛋白浓度依赖关系，与对照组及其它各实验组相比有极显著性差异(P＜0.001)，E14和E18 CE分别在蛋白浓度为0.20和0.40 mg/ml时有最大神经营养活性。结论：CE对培养脊髓神经细胞具有明显营养作用。
关键词 靶源性神经营养因子；小脑提取液；胎龄；脊髓
Effect of Cerebellum Extracts from Embryonic Rat on Cultured Spinal Neural Cells
L Jie, L Yongli
Neuroanatomy Research Office, Brain Institute, China Medical University, Shenyang, 110001
ABSTRACT Objective:The effects of cerebellum extracts (CE) from different embryonic aged of rat on the cultured rat spinal cells were studied. Methods：CE from 14 d, 16 d, 18 d, 20 d aged of embryonic rats (E14～E20) were collected and were made into solutions of containing different concentrations of proteins. Then these solutions were added to the cultured spinal cells of E18 rats. The neuronal survival was determined by the auto-colorimetric assay. Results:CE of all experimental aged of rats could promote the survival of spinal neurons, and there was also a concentration-dependent relation on protein, with significant differences (P<0.001) between the control and experimental groups. The most prominent neurotrophic activities were found in E14 CE of 0.20 mg/ml and E18 CE of 0.40 mg/ml. Conclusion:CE has significant neurotrophic effects on cultured spinal neural cells.
KEY WORDS target-derived neurotrophic factors; cerebellum extracts; embryonic age; spinal cord
受损伤神经元的存活及功能恢复有赖于这些神经元的靶细胞提供的特异的靶源性神经营养因子(target-derived neurotrophic factors, TNTFs)［1］。小脑是脊髓的主要靶器官之一，已发现有多种神经营养因子(neurotrophic factors NTFs)及其受体在两者表达，但以小脑为靶器官提供的TNTFs还未见报道。胚胎小脑是一种在脑移植中被广为应用的移植物，它能抑制瘢痕组织生长，促进神经组织再生［2］，诱导损伤后脊髓小脑束的重建［1］。为进一步探讨胚胎小脑中的有效神经营养成分，我们制备了小脑提取液(cerebellum extracts, CE)，观察了它对培养脊髓细胞的作用。
1 材料与方法
1.1 CE的制备
取相应胎龄(E14、E16、E18、E20)胚鼠小脑各20个，制备匀浆，经超声粉碎，超速离心，取上清液，以紫外分光光度法测定蛋白质浓度，以PBS (pH7.0[1] [2] [3] 下一页
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