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目的:明确Lhx8在SD大鼠切割穹窿海马伞后海马齿状回颗粒下层和门区中的表达变化。方法:切割SD大鼠穹窿海马伞,成功制备海马去神经支配动物模型后,通过Western Blot的方法检测切割后第1、3、7、14、21、28 d海马中Lhx8蛋白的表达变化;通过免疫组织化学方法检测切割后第7天海马齿状回颗粒下层和门区中Lhx8阳性细胞的表达定位,并通过免疫荧光化学方法检测Lhx8/ChAT双标阳性细胞的共定位情况;切割SD大鼠穹窿海马伞并制备海马提取液,将其加入至细胞培养液中,模拟体内海马神经再生微环境,检测培养的海马放射状胶质细胞(radial slia cells,RGCs)向胆碱能神经元分化的情况。结果:切割穹窿海马伞后第7 d,Lhx8蛋白表达量较其他各时相点明显增高;海马齿状回颗粒下层和门区中Lhx8阳性细胞数目和光密度也明显高于正常侧,Lhx8/ChAT双标阳性细胞数目也较正常侧增多;体外细胞培养,切割穹窿海马伞侧海马提取液也较正常侧海马提取液更能促进海马放射状胶质细胞向胆碱能神经元的分化。结论:切割穹窿海马伞后,Lhx8表达明显上调,与海马齿状回的胆碱能神经再生有关。
OBJECTIVE: To determine the expression of Lhx8 in the inferior layer and the portal area of the hippocampal dentate gyrus after the Fn is cut in SD rats. Methods: The SD rat FNF was excised and the hippocampus denervated animal model was successfully established. The expression of Lhx8 protein in hippocampus was detected by Western Blot after 1, 3, 7, 14, 21, Immunohistochemistry was used to detect the expression of Lhx8-positive cells in the lower layer and the portal area of hippocampal dentate gyrus on the 7th day after cleavage. The co-localization of Lhx8 / ChAT double positive cells was detected by immunofluorescence staining. Fimbria fornix and prepare the hippocampus extract, which was added to the cell culture medium to simulate the hippocampal nerve regeneration microenvironment in vivo, to detect the differentiation of the cultured hippocampal radial slia cells (RGCs) into the cholinergic neurons . Results: On the 7th day after fimbria fornix transection, the expression of Lhx8 protein was significantly increased compared with other time points. The numbers and optical density of Lhx8 positive cells in the inferior layer and the portal area of hippocampal dentate gyrus were also significantly higher than those in normal Lhx8 / ChAT Double positive cells also increased compared with the normal side; In vitro cell culture, hippocampal fimbria for cutting hippocampal extracts than normal hippocampal extracts more can promote hippocampal radial glial cells to cholinergic neurons differentiation. Conclusion: Lnx8 expression is significantly up-regulated after Fimbria fornix transection, which is related to the cholinergic regeneration in hippocampal dentate gyrus.