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为探讨切割穹窿海马伞海马提取液和银杏叶提取物(extract of ginkgo bilobo,EGb)在海马NSCs向胆碱能神经元定向分化中的作用,分别制备大鼠穹窿海马伞切割侧和正常侧海马提取液;将从鼠胚海马中分离扩增的NSCs球分成4组,应用不同的培养液促其分化:(1)联合组:含切割侧海马提取液和银杏内酯的DMEM/F12培养基;(2)提取液组:含切割侧海马提取液的DMEM/F12培养基;(3)EGb组:含银杏内酯的DMEM/F12培养基;(4)对照组:含正常侧海马提取液的DMEM/F12培养基。培养14d后行ChAT免疫荧光检测,计算ChAT阳性神经元的分化率,图像处理细胞面积和周长。结果显示联合组各项指标均明显优于其它各组(P<0.01);提取液组、EGb组各项指标也均优于对照组(P<0.05);细胞面积提取液组优于EGb组(P<0.05),细胞周长EGb组优于提取液组(P<0.05),两组ChAT阳性神经元分化率无明显差异(P>0.05)。上述提示切割穹窿海马伞的海马提取液和EGb联合应用可诱导海马NSCs分化为更多、更为成熟的胆碱能神经元。
To investigate the role of cutting hippocampi hippocampus extract and extract of ginkgo bilobo (EGb) in the differentiation of hippocampal NSCs into cholinergic neurons, the hippocampi of the rat hippocampus were prepared. Extracts; NSCs isolated from rat embryonic hippocampus were divided into 4 groups and cultured in different media to promote their differentiation: (1) Combined group: DMEM/F12 medium containing cut hippocampal extract and ginkgolide (2) Extract group: DMEM/F12 medium containing cut side hippocampus extract; (3) EGb group: DMEM/F12 medium containing ginkgo lactone; (4) Control group: hippocampus extract containing normal side DMEM/F12 medium. After 14 days of culture, ChAT immunofluorescence was performed to determine the differentiation rate of ChAT-positive neurons and the area and perimeter of the image-processing cells. The results showed that the indexes in the combination group were significantly better than those in other groups (P<0.01); the indexes in the extract group and the EGb group were also better than those in the control group (P<0.05); the cell area extract group was superior to the EGb group. (P<0.05), the cell perimeter EGb group was superior to the extract group (P<0.05), and there was no significant difference in the differentiation rate of ChAT-positive neurons between the two groups (P>0.05). The combined use of hippocampal extract and EGb for cutting the temporal lobe hippocampus can induce hippocampal NSCs to differentiate into more mature cholinergic neurons.