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目的探讨切割穹隆海马伞大鼠海马胆碱能神经再生过程中Lhx8的作用。方法 72只SD大鼠随机分成4组,每组18只。所分的4组分别为:对照组(未做任何处理),切割组(切割右侧穹隆海马伞),过表达组(右侧海马齿状回中注射Lenti6.3-Lhx8慢病毒)及切割联合过表达组(右侧海马齿状回中注射Lenti6.3-Lhx8慢病毒并切割右侧穹隆海马伞)。28d后每组各取6只分别制备脑冷冻切片行免疫荧光检测,提取海马总RNA行Real-time PCR,提取海马蛋白行Western blotting检测。结果与对照组相比,切割组、过表达组和切割联合过表达组中Lhx8 mRNA和蛋白水平均显著上调,以切割联合过表达组中上调最为显著,组间比较差异均有统计学意义;切割组胆碱乙酰转移酶(ChAT)mRNA水平与对照组相比无统计学差异,过表达组和切割联合过表达组中ChAT mRNA显著上调,且以切割联合过表达组中上调最为显著;切割组、过表达组和切割联合过表达组中均检测到ChAT蛋白,组间比较差异有统计学意义;切割组、过表达组和切割联合过表达组海马齿状回门区和颗粒下层中均能检测到新生的Lhx8阳性的胆碱能神经元,且以切割联合过表达组最多,组间比较差异均有统计学意义。结论上调Lhx8的表达能促进海马中神经干细胞向胆碱能神经元分化,Lhx8可能与海马胆碱能神经再生有关。
Objective To investigate the role of Lhx8 in hippocampal cholinergic nerve regeneration in the sedentary FHL rats. Methods 72 SD rats were randomly divided into 4 groups with 18 rats in each group. The four groups were divided into control group (without any treatment), cutting group (right dome FNA), overexpression group (Lenti6.3-Lhx8 lentivirus injected into the right hippocampal dentate gyrus) The co-overexpression group (Lenti 6.3-Lhx8 lentivirus injected into the right hippocampal dentate gyrus and the right dome of the hippocampus was excised). After 28 days, 6 rabbits in each group were separately prepared for immunofluorescence detection of brain frozen sections. Real-time PCR was used to extract the total RNA of hippocampus, Western blotting was used to detect the protein in hippocampus. Results Compared with the control group, the expression of Lhx8 mRNA and protein in the overexpression group and the overexpression group were significantly up-regulated. The up-regulation was most significant in the combined overexpression group and the differences between the two groups were statistically significant. There was no significant difference in the level of ChAT mRNA between the two groups (P <0.05). Compared with the control group, there was no significant difference in the level of ChAT mRNA in the control group. ChAT mRNA was significantly up-regulated in the overexpression group and the cleaved overexpression group, ChAT protein was detected in the overexpression group, overexpression group and combined overexpression group, and there was significant difference between the two groups. In the cutting group, overexpression group and cutting combined overexpression group, Neonatal Lhx8-positive cholinergic neurons were detectable, and the co-expression group was the most common in cutting and combining groups with statistically significant differences. Conclusion Upregulation of Lhx8 can promote the differentiation of neural stem cells into cholinergic neurons in the hippocampus. Lhx8 may be related to cholinergic neurogenesis in the hippocampus.