目的:通过对小鼠进行24 h束缚应激构建快速抑郁样行为模型,探究DNA甲基化调控BDNF表达在小鼠抑郁行为中发挥的作用。方法:使用8～10周龄体质量20～22 g的ICR雄性小鼠作为实验对象,随机分为两组,即正常组(Control)和抑郁组(Restraint),通过行为学检测小鼠是否产生抑郁行为,并检测小鼠海马脑区脑源性神经营养因子(Brain-derived neurotrophic factor,BDNF)及其受体TrkB的mRNA及蛋白表达变化。检测小鼠DNA甲基转移酶1(DNMT1)、DNA甲基转移酶3A(DNMT3A)、DNA甲基转移酶3B(DNMT3B)的mRNA表达变化以及焦磷酸测序检测小鼠BDNF启动子区域甲基化水平。结果:与Control组相比,Restriant组的移动总距离缩短(P＜0.05);与Control组相比,Restriant组的糖水偏好比率下降(P＜0.05);Restriant组的体质量变化与Control组相比,其体质量恢复的幅度下降(P＜0.05);与Control组相比,Restriant组BDNF的mRNA和蛋白的表达降低(P＜0.05),TrkB的蛋白及mRNA表达差异无统计学意义(P＞0.05);DNMT3A、DNMT3B的mRNA表达升高(P＜0.05),DNMT1的mRNA表达差异无统计学意义;与Control组相比,Restriant组的BDNF启动子区域DNA甲基化程度明显升高(P＜0.05)。结论:小鼠经过24 h束缚应激后,可诱导小鼠产生明显的抑郁样行为,这可能是通过上调DNA甲基化水平进而下调BDNF表达而发挥作用。
Objective: To explore the molecular mechanism of DNA methylation regulation of BDNF expression in depressive behavior of mice by establishing a rapid depression-like behavior model in mice under 24-hour restraint stress. Methods: ICR male mice aged 8-10 weeks weighing 20-22g were used as experimental subjects. They were randomly divided into two groups, namely the normal group (Control) and the depression group (Restraint). The depressive behavior of mice was detected by behavior, and the mRNA and protein expression of brain-derived neurotrophic factor (BDNF) and its receptor TrkB in hippocampus were detected. The mRNA expression of DNA methyltransferase 1, DNA methyltransferase 3A, DNA methyltransferase 3B (DNMT1, DNMT3A, DNMT3B) in mice was detected, and the methylation level of BDNF promoter region in mice was detected by pyrosequencing. Results: Compared with the Control group, the total distance of movement in the Restriant group was shortened (P＜0.05), the sugar water preference ratio of the Restriant group decreased (P＜0.05), the body weight recovery of the Restriant group decreased (P＜0.05), the expression of BDNF mRNA and protein in the Restriant group was decreased (P＜0.05), but there was no significant difference in the expression of TrkB protein and mRNA between the Control group and the Restriant group (P＞0.05). And compared with the Control group, the mRNA expression of DNMT3 A and DNMT3 B increased (P＜0.05), and there was no significant difference in the mRNA expression of DNMT1 between the Control group and the Restriant group. Compared with the Control group, the degree of DNA methylation in the BDNF promoter region of the Restriant group was significantly increased (P＜0.05). Conclusion: After 24 h restraint stress, mice can induce obvious depression-like behavior, which may play a role by up-regulating DNA methylation level and down-regulating BDNF expression.
[1] Chu XX, Zhou Y, Hu ZQ, et al. 24-hour-restraint stress induces long-term depressive-like phenotypes in mice[J]. Sci Rep, 2016,6:32935.
[2] Ma H, Li JF, Qiao X, et al. Sigma-1 receptor activation mediates the sustained antidepressant effect of ketamine in mice via increasing BDNF levels[J]. Acta Pharmacol Sin, 2023:1-10.
[3] Ye ZY, Wang JW, Fang F, et al. Zhi-Zi-Hou-Po decoction alleviates depressive-like behavior and promotes hippocampal neurogenesis in chronic unpredictable mild stress induced mice via activating the BDNF/TrkB/CREB pathway[J]. J Ethnopharmacol, 2024,319(Pt 3):117355.
[4] 张世纪,付翁,贾小娥,等. 5-Aza-cdR通过DNA甲基化调节BDNF参与学习记忆的机制[J]. 包头医学院学报,2022,38(1):21-25.
[5] Zhang SJ, Fu W, Jia XE, et al. Hypoxic preconditioning modulates BDNF and its signaling through DNA methylation to promote learning and memory in mice[J]. ACS Chem Neurosci, 2023,14(12):2320-2332.
[6] Liu FL, Huang SH, Guo D, et al. Deep brain stimulation of ventromedial prefrontal cortex reverses depressive-like behaviors via BDNF/TrkB signaling pathway in rats[J]. Life Sci, 2023,334:122222.
[7] 储锡霞. 环境应激模式动物的建立及其神经生物学机制的研究[D]. 上海:上海交通大学,2017.
[8] Yao CH, Zhang YW, Sun XR, et al. Areca catechu L. ameliorates chronic unpredictable mild stress-induced depression behavior in rats by the promotion of the BDNF signaling pathway[J]. Biomed Pharmacother, 2023,164:114459.
[9] Duman RS, Deyama S, Fogaça MV. Role of BDNF in the pathophysiology and treatment of depression: activity-dependent effects distinguish rapid-acting antidepressants[J]. Eur J Neurosci, 2021,53(1):126-139.
[10] Tang YH, Su H, Nie KX, et al. Berberine exerts antidepressant effects in vivo and in vitro through the PI3K/AKT/CREB/BDNF signaling pathway[J]. Biomed Pharmacother, 2024,170:116012.
[11] Xu YY, Zhang J, Yu LY, et al. Engeletin alleviates depression-like phenotype by increasing synaptic plasticity via the BDNF-TrkB-mTORC1 signalling pathway[J]. J Cell Mol Med, 2023,27(23):3928-3938.
[12] Zeng JS, Xie ZQ, Chen L, et al. Rosmarinic acid alleviate CORT-induced depressive-like behavior by promoting neurogenesis and regulating BDNF/TrkB/PI3K signaling axis[J]. Biomed Pharmacother, 2024,170:115994.
[13] Zhao T, Piao LH, Li DP, et al. BDNF gene hydroxymethylation in hippocampus related to neuroinflammation-induced depression-like behaviors in mice[J]. J Affect Disord, 2023,323:723-730.
[14] Sales AJ, Maciel IS, Suavinha ACDR, et al. Modulation of DNA methylation and gene expression in rodent cortical neuroplasticity pathways exerts rapid antidepressant-like effects[J]. Mol Neurobiol, 2021,58(2):777-794.
[15] Vidovi Č E, Pelikan S, Atanasova M, et al. DNA methylation patterns in relation to acute severity and duration of anxiety and depression[J]. Curr Issues Mol Biol, 2023,45(9):7286-7303.
