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目的研究APP5肽对体外培养的大鼠胚胎海马神经干细胞(NSCs)增殖和分化的影响。方法1.原代培养SD大鼠胚胎脑海马NSCs;2.利用5-溴脱氧尿嘧啶核苷(BrdU)和神经元、星型胶质细胞、少突胶质细胞的特异性标记物微管相关蛋白2(MAP2)、胶质纤维酸性蛋白(GFAP)、半乳糖脑苷脂(GalC)对培养的NSCs进行鉴定;3.将培养的NSCs分为对照组、血清组、APP5肽反序列组和APP5肽组,观察各组细胞形态的变化;4.将培养的NSCs分为对照组、APP5肽反序列组和APP5肽组,利用细胞计数、干细胞克隆形成率、干细胞克隆形成大小以及四甲基偶氮唑盐(MTT)代谢率测定的方法,分析APP5肽对海马NSCs增殖的影响。结果1.海马神经干细胞呈神经球聚集生长,BrdU染色阳性;加入血清后神经球周围有细胞呈放射状向四周生长,并带有突起。染色呈MAP2、GFAP或GalC阳性;2.海马NSCs加入APP5肽及其反序列后细胞形态与对照组相比没有明显改变;3.与对照组相比,加APP5肽后海马NSCs数量明显增加,克隆形成率和克隆形成的直径均有明显的增加,并有统计学差异;4.与对照组相比,加入5肽后海马NSCs的MTT代谢率明显增加,有统计学差异,反序列组没有明显改变。结论APP5肽能促进海马NSCs增殖,但并不促进其分化。
Objective To study the effect of APP5 peptide on the proliferation and differentiation of rat embryonic hippocampal neural stem cells (NSCs) cultured in vitro. Methods 1. Primary cultured SD rat embryonic hippocampal NSCs; 2.Using 5-bromodeoxyuridine (BrdU) and neurons, astrocytes, oligodendrocyte-specific markers microtubules NSCs were identified as MAP2, GFAP and GalC.3. The cultured NSCs were divided into control group, serum group, APP5 peptide reverse sequence group And APP5 peptide group to observe the morphological changes of each group.4. The cultured NSCs were divided into control group, APP5 peptide reverse sequence group and APP5 peptide group. Using the cell counting, the rate of stem cell clone formation, the size of stem cell clone, The effect of APP5 peptide on the proliferation of hippocampal NSCs was assayed by MTT metabolic rate assay. The neurons in hippocampal neurons gathered and grew, BrdU staining was positive. After the addition of serum, the cells around the neurospheres grew radially and had protrusions. The staining of MAP2, GFAP or GalC was positive; 2. The morphological changes of NSCs in hippocampus after adding APP5 peptide and its reverse sequence did not change significantly compared with the control group; 3. Compared with the control group, the number of NSCs in hippocampus increased significantly after adding APP5 peptide, The formation rate of colony and the diameter of colony formation increased obviously, and there was statistical difference.4.Compared with control group, the MTT metabolic rate of hippocampal NSCs increased obviously after addition of 5-peptide, and the difference was significant Significant change. Conclusion APP5 peptide can promote the proliferation of hippocampal NSCs but does not promote their differentiation.