miR-124通过促进海马神经干细胞增殖分化改善MK-801介导的小鼠焦虑样行为

doi:10.12122/j.issn.1673-4254.2026.08.15

摘要/Abstract

摘要：

目的 探究 miR-124对MK-801介导小鼠焦虑样行为的调控作用。方法将60只6周龄C57BL/6J雄性小鼠随机分为CON组、MK-801+Saline组、MK-801+Vehicle组和MK-801+miR-124组，每组15只。通过腹腔注射MK-801诱导焦虑样模型，脑立体定位注射mmu-miR-124-3p mimics以过表达miR-124，RT-qPCR实验检测miR-124是否成功转染，行为学实验检测miR-124对小鼠焦虑样行为的影响，Western blotting实验检测miR-124对小鼠海马神经元蛋白表达的影响，免疫荧光染色实验检测miR-124对神经干细胞数量、新增殖神经元数量及神经元分化的影响。结果 miR-124上调后海马miR-124水平显著升高（P<0.05），小鼠旷场实验中总活动距离（P<0.05）与中央区活动距离（P<0.001）、高架十字迷宫实验中开放臂活动时间百分比（P<0.05）均显著升高，小鼠海马Tuj1蛋白表达量显著增加（P<0.001），小鼠海马神经干细胞数量和新生神经元数量均显著增多（P<0.001），新生神经元与成熟神经元荧光面积比值亦显著增加（P<0.001）。结论 miR-124可通过促进海马神经干细胞增殖分化、增加神经元数量，改善MK-801介导的小鼠焦虑样行为，为焦虑症治疗提供潜在靶点。

关键词: miR-124, 焦虑, 神经干细胞, 神经元

Abstract:

Objective To investigate the regulatory effect of miR-124 on MK-801-induced anxiety-like behavior in mice. Methods Sixty 6-week-old male C57BL/6J mice were randomly divided into control group, MK-801+Saline group, MK-801+Vehicle group, and MK-801+miR-124 group (n=15). Anxiety-like models were established in the latter 3 groups by intraperitoneal injection of MK-801, and stereotaxic brain injection of mmu-miR-124-3p mimics was performed for local miR-124 overexpression in the hippocampus, and successful miR-124 transfection was verified using RT-qPCR. Behavioral tests were conducted to assess the effect of miR-124 overexpression on anxiety-like behaviors of the mice. Western blotting and immunofluorescence staining were used to examine the effects of miR-124 overexpression on hippocampal neuronal protein expressions, neural stem cell number, number of new neurons, and neuronal differentiation. Results The mice in MK-801+miR-124 group showed significantly increased hippocampal miR-124 levels after the injection. The mice receiving miR-124 microinjection exhibited significant increases in both the total and central zone distance traveled in open field test and in percentages of time spent in the open arms in elevated plus maze test. Hippocampal Tuj1 protein expression and the numbers of hippocampal neural stem cells and newborn neurons, along with the fluorescence area ratio of newborn to mature neurons, increased significantly in the mice in MK-801+miR-124 group. Conclusion miR-124 overexpression ameliorates MK-801-induced anxiety-like behaviors in mice by promoting hippocampal neural stem cell proliferation and differentiation and increasing hippocampal neuron number, suggesting the value of miR-124 as a potential therapeutic target for anxiety disorders.

Key words: miR-124, anxiety, neural stem cells, neurons

余东焜, 支仕杰, 刘彦波, 杨治伦, 刘娟, 邵钰. miR-124通过促进海马神经干细胞增殖分化改善MK-801介导的小鼠焦虑样行为[J]. 南方医科大学学报, 2026, 46(8): 1878-1885.

Dongkun YU, Shijie ZHI, Yanbo LIU, Zhilun YANG, Juan LIU, Yu SHAO. MiR-124 ameliorates MK-801-induced anxiety-like behaviors in mice by promoting hippocampal neural stem cell proliferation and differentiation[J]. Journal of Southern Medical University, 2026, 46(8): 1878-1885.

图/表 6

图1 RT-qPCR验证病毒转染

Fig.1 RT-qPCR validation of viral transfection. CON: Control group; MK-801: Dizocilpine; Vehicle: Adeno-associated Virus Serotype 9-Glycoprotein-2-Negative Control; miR-124: Mus musculus microRNA-124-3p mimics. *P<0.05 vs CON group; #P<0.05 vs MK-801+Saline group (n=3).

图2 行为学实验检测miR-124对小鼠焦虑样行为的改善情况

Fig.2 Behavioral tests for assessing the ameliorative effects of miR-124 on anxiety-like behaviors in mice. A: Trajectory diagram in open field test. B: Trajectory diagram in elevated cross maze test. C: Comparison of distance traveled in the center region in open field test. D: Comparison of total distance traveled in open field test. E: Comparison of percentage of time in the open arm in the elevated cross maze test. *P<0.05, **P<0.01, ***P<0.001 vs CON group; #P<0.05, ##P<0.01, ###P<0.001 vs MK-801+Saline group (n=8).

图3 miR-124逆转MK-801抑制Tuj1蛋白的作用

Fig.3 miR-124 reverses the inhibitory effect of MK-801 on Tuj1 protein. A: Tuj1 protein bands in Western blotting. B: Tuj1 protein expression levels in the 4 groups. ***P<0.001 vs CON group; ###P<0.001 vs MK-801+Saline group (n=6).

图4 miR-124逆转MK-801抑制神经干细胞增殖的作用

图5 miR-124逆转MK-801抑制新生神经元增殖的作用

图6 miR-124逆转MK-801抑制小鼠DCX与NeuN正常比例的作用

Fig.6 miR-124 reverses the inhibitory effect of MK-801 on DCX to NeuN ratio in the mice (Scale bar=100 μm). A: DCX and NeuN fluorescence staining. B: DCX and NeuN ratio statistics. ****P<0.0001 vs CON group; ####P<0.0001 vs MK-801+Saline group; Zoom in the image to 5 times its original size (n=9).

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