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Esketamine Improves Working Memory Impairment in Neuropathic Mice Through Hippocampal BDNF-TrkB Pathway
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摘要:
目的 探究艾司氯胺酮改善神经病理性疼痛所致小鼠工作记忆障碍的作用及可能机制。 方法 将50只2月龄雄性C57BL/6J小鼠采用随机数字表法均分为5组：假手术+生理盐水组(SN组)、慢性坐骨神经压迫(chronic constriction injury，CCI)+生理盐水组(CN组)、CCI+艾司氯胺酮组(CE组)、CCI+ANA-12组(CA组)、CCI+ANA-12+艾司氯胺酮组(CAE)组，每组10只。采用CCI建立神经病理性疼痛模型，于造模后第16天，CE组、CAE组分别给予艾司氯胺酮10 mg/kg，其中CAE组于艾司氯胺酮注射前30 min给予ANA-12(一种选择性TrkB受体拮抗剂)0.5 mg/kg；CA组仅给予ANA-12；SN组、CN组仅给予等量生理盐水，连续5 d腹腔注射给药。于造模后第21天行旷场实验、Y迷宫实验，检测小鼠机械缩足反应阈值(paw withdrawal threshold，PWT)及热缩足潜伏期(paw withdrawal latency，PWL)，并于造模后第21~23天腹腔注射溴脱氧尿嘧啶核苷(Bromo-2-deoxyUridine，BrdU)。通过蛋白质免疫印迹试验检测小鼠海马脑源性神经营养因子(brain-derived neurotrophic factor，BDNF)蛋白表达，采用免疫荧光检测海马齿状回BrdU及双皮质素(doublecortin，DCX)共染阳性(BrdU+/DCX+)细胞数量。 结果 与SN组相比，CN组、CE组、CA组、CAE组PWT明显降低(P均＜0.05)，PWL明显缩短(P均＜0.05)；在旷场实验中，5组小鼠的运动总距离无明显差异(P=0.142)；Y迷宫实验中，与SN组相比，CN组自发性交替正确率明显降低(P＜0.001)，与CN组相比，CE组自发性交替正确率明显升高(P＜0.001)，与CE组相比，CAE组自发性交替正确率明显降低(P=0.004)；与SN组相比，CN组BDNF表达下调(P=0.021)，与CN组相比，CE组BDNF表达上调(P=0.030)，与CE组相比，CAE组表达下调(P=0.043)；与SN组相比，CN组BrdU+/DCX+细胞数量明显降低(P=0.025)，与CN组相比，CE组BrdU+/DCX+细胞数量明显升高(P=0.003)，与CE组相比，CAE组BrdU+/DCX+细胞数量明显降低(P=0.014)。 结论 艾司氯胺酮可能通过海马BDNF-TrkB神经通路促进海马齿状回神经再生，进而改善神经病理性疼痛所致的工作记忆障碍。 Abstract:Objective To explore the effect of esketamine on working memory impairment in neuropathic mice and its underlying mechanism. Methods Fifty clean grade male C57BL/6J mice (2 months) were divided into 5 groups by random number table method: sham+saline (SN group), CCI+saline (CN group), CCI+esketamine(CE group), CCI+ANA-12 (CA group), CCI+ANA-12+esketamine (CAE group), with 10 mice in each group. Chronic constriction injury (CCI) was employed to establish a neuropathic pain model. On the 16th day after modeling, CE group and CAE group were administered ketamine (10 mg/kg), CAE group received ANA-12 (0.5 mg/kg) half an hour before ketamine injection, CA group was only given ANA-12, and SN group and CN group received an equivalent volume of saline. The administration was done through intraperitoneal injection for 5 consecutive days. The open-field test (OFT), paw withdrawal threshold (PWT), paw withdrawal latency (PWL) and Y-maze test were performed from day 21 after surgery. Bromo-2-deoxyUridine (BrdU) was dissolved in saline and intraperitoneally injected into the mice on days 21 to 23 after the surgery. Western blot was performed to determine the expression of Brain-derived neurotrophic factor(BDNF) in hippocampus, the immunofluorescence was performed to determine the number of BrdU and doublecortxin (DCX) positive cells in the dentate gyrus (DG) area of hippocampal. Results Compared with SN group, the other four groups showed significant reductions in both PWT and PWL on day 21 after surgery (all P<0.05); There was no significant difference in the total distance travelled by the 5 groups of mice (P=0.142); In the Y maze test, compared to SN group, the accurate percentage of spontaneous alternation in CN group showed significant reductions (P<0.001), which was reversed by esketamine administration (P<0.001); compared with CE group, there was a significant reduction of the accurate percentage of spontaneous alternation in CAE group (P=0.004). The expression of BDNF protein in CN group was lower than that in SN group (P=0.021) and CE group (P=0.030), and compared with CE group, the expression of BDNF was significant decreased in CEA group (P=0.043). The number of BrdU positive and DCX positive cells significantly reduced in the DG area of the hippocampus in CN group compared to SN group (P=0.025) and CE group (P=0.003). The number of BrdU positive and DCX positive cells in CAE group significantly reduced in the DG region of the hippocampus compared to CE group (P=0.014). Conclusion Esketamine maybe improve working memory impairment in neuropathic mice and the neurogenesis in dentate gyrus area of hippocampal through the BDNF-TrkB pathway. -
Key words:
- esketamine /
- neuropathic pain /
- working memory /
- hippocampus
作者贡献：蒋玉斌、王星明负责研究实施及论文撰写；周志强、杨建军负责研究设计并指导论文修订；蒋玉斌、王星明、张跃负责实验设计、数据分析及结果解读；王星明负责实验过程及文献查询。利益冲突：所有作者均声明不存在利益冲突 -
图 1 5组小鼠海马BDNF相对表达水平(n=5)
A. BDNF及β-actin蛋白免疫印迹；B. BDNF相对表达量
Figure 1. Expression of hippocampal BDNF in 5 groups (n=5)
A. Western blot; B. Relative expression levels of hippocampal BDNF
图 2 5组小鼠BrdU+/DCX+在海马齿状回的表达水平(n=5)
A. 免疫荧光检测(20×)；B. BrdU+/DCX+细胞数目
Figure 2. Expression of BrdU+/DCX+in the dentate gyrus (n=5)
A. Immunofluorescence(20×); B. The number of BrdU+/DCX+cells in the dentate gyrus
表 1 5组小鼠PWT和PWL比较(x±s)
Table 1. Comparison of PWT and PWL among 5 groups (x±s)
组别 PWT(g) PWL(s) SN组(n=10) 0.804±0.226 11.25±2.91 CN组(n=10) 0.086±0.094* 4.80±1.42* CE组(n=10) 0.181±0.189* 5.99±1.51* CA组(n=10) 0.064±0.036* 4.47±1.18* CAE组(n=10) 0.069±0.076* 4.35±0.86* PWT(paw withdrawal threshold)：机械缩足反射阈值；PWL(paw withdrawal latency)：热缩足潜伏期；与SN组比较，*P＜0.001 
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表 2 5组小鼠旷场实验结果比较(x±s)
Table 2. Comparison of open field test results among 5 groups (x±s)
组别 进入中央区次数(次) 运动总距离(mm) SN组(n=10) 14.10±6.49 14892.10±3843.94 CN组(n=10) 13.55±7.20 12292.35±4317.10 CE组(n=10) 18.18±5.91 14518.19±3969.52 CA组(n=10) 14.73±5.26 12801.53±3969.52 CAE组(n=10) 19.00±4.85 16371.60±2664.84 F值 2.129 1.811 P值 0.092 0.142
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表 3 5组小鼠进臂总次数及自发性交替正确率比较(x±s)
Table 3. The total number of arm entries and spontaneous alternations among 5 groups (x±s)
组别 进臂总次数(次) 自发性交替正确率(%) SN组(n=10) 23.70±3.06 72.41±7.43 CN组(n=10) 22.90±2.85 54.21±7.27* CE组(n=10) 23.10±2.13 70.70±4.83# CA组(n=10) 23.00±4.37 57.75±6.28 CAE组(n=10) 22.90±2.51 59.59±10.65† F值 0.118 11.26 P值 0.975 0.001 CN组与SN组比较，*P＜0.001；CN组与CE组比较，#P＜0.001；CAE组与CE组比较，†P=0.004
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[1] Mazza S, Frot M, Rey A E. A comprehensive literature review of chronic pain and memory[J]. Prog Neuropsychopharmacol Biol Psychiatry, 2018, 87(Pt B): 183-192. [2] Miller E K, Lundqvist M, Bastos A M. Working memory 2.0[J]. Neuron, 2018, 100(2): 463-475. doi: 10.1016/j.neuron.2018.09.023 [3] Berryman C, Stanton T R, Jane Bowering K, et al. Evid-ence for working memory deficits in chronic pain: a systematic review and meta-analysis[J]. Pain, 2013, 154(8): 1181-1196. doi: 10.1016/j.pain.2013.03.002 [4] Zhang G F, Zhou Z Q, Guo J, et al. Histone deacetylase 3 in hippocampus contributes to memory impairment after chronic constriction injury of sciatic nerve in mice[J]. Pain, 2021, 162(2): 382-395. doi: 10.1097/j.pain.0000000000002056 [5] Wei Y, Chang L J, Hashimoto K. Molecular mechanisms underlying the antidepressant actions of arketamine: beyond the NMDA receptor[J]. Mol Psychiatry, 2022, 27(1): 559-573. doi: 10.1038/s41380-021-01121-1 [6] Wei Y, Chang L J, Hashimoto K. A historical review of antidepressant effects of ketamine and its enantiomers[J]. Pharmacol Biochem Behav, 2020, 190: 172870. doi: 10.1016/j.pbb.2020.172870 [7] Zhang C, He J Q, Shi Q Y, et al. Subanaesthetic dose of esketamine during induction delays anaesthesia recovery a randomized, double-blind clinical trial[J]. BMC Anesthesiol, 2022, 22(1): 138. doi: 10.1186/s12871-022-01662-0 [8] Hashimoto K. Arketamine for cognitive impairment in psychiatric disorders[J]. Eur Arch Psychiatry Clin Neurosci, 2023, 273(7): 1513-1525. doi: 10.1007/s00406-023-01570-5 [9] Zhang K, Yao Y T, Hashimoto K. Ketamine and its metabolites: potential as novel treatments for depression[J]. Neuropharmacology, 2023, 222: 109305. doi: 10.1016/j.neuropharm.2022.109305 [10] Xia S H, Hu S W, Ge D G, et al. Chronic pain impairs memory formation via disruption of neurogenesis mediated by mesohippocampal brain-derived neurotrophic factor signaling[J]. Biol Psychiatry, 2020, 88(8): 597-610. doi: 10.1016/j.biopsych.2020.02.013 [11] Wu A B, Zhang J Y. Neuroinflammation, memory, and depression: new approaches to hippocampal neurogenesis[J]. J Neuroinflammation, 2023, 20(1): 283. doi: 10.1186/s12974-023-02964-x [12] Wang X M, Zhang G F, Jia M, et al. Environmental enrichment improves pain sensitivity, depression-like phenotype, and memory deficit in mice with neuropathic pain: role of NPAS4[J]. Psychopharmacology (Berl), 2019, 236(7): 1999-2014. doi: 10.1007/s00213-019-5187-6 [13] Hussain G, Akram R, Anwar H, et al. Adult neurogenesis: a real hope or a delusion?[J]. Neural Regen Res, 2024, 19(1): 6-15. doi: 10.4103/1673-5374.375317 [14] Jaberi S, Fahnestock M. Mechanisms of the beneficial effects of exercise on brain-derived neurotrophic factor expression in Alzheimer's disease[J]. Biomolecules, 2023, 13(11): 1577. doi: 10.3390/biom13111577 [15] 李亚南, 张琦, 于家旭, 等. BDNF/TrkB信号通路在预先注射青年大鼠血浆减轻七氟烷诱发老龄大鼠认知功能障碍中的作用[J]. 中华麻醉学杂志, 2022, 42(5): 546-550. Li Y N, Zhang Q, Yu J X, et al. Role of BDNF/TrkB signaling pathway in pre-injection of young rat plasma-induced reduction of sevoflurane-caused cognitive dysfunc-tion in aged rats[J]. Chin J Anesthesiol, 2022, 42(5): 546-550. [16] Fabrazzo M, Cipolla S, Pisaturo M, et al. Bidirectional relationship between HIV/HBV infection and comorbid depres-sion and/or anxiety: a systematic review on shared biological mechanisms[J]. J Pers Med, 2023, 13(12): 1689. doi: 10.3390/jpm13121689 [17] Pettorruso M, Miuli A, Clemente K, et al. Enhanced peripheral levels of BDNF and proBDNF: elucidating neurotrophin dynamics in cocaine use disorder[J/OL]. Mol Psychiatry. https://doi.org/10.1038/s41380-023-02367-7 . doi:10.1038/s41380-023-02367-7 .[18] Zhang S X, Chen Y R, Wang Y J, et al. Tau accumulation in the spinal cord contributes to chronic inflammatory pain by upregulation of IL-1β and BDNF[J/OL]. Neurosci Bull. https://doi.org/10.1007/s12264-023-01152-4 . doi:10.1007/s12264-023-01152-4 .[19] Cook A A, Leung T C S, Rice M, et al. Endosomal dysfunction contributes to cerebellar deficits in spinocerebellar ataxia type 6[J]. Elife, 2023, 12: RP90510. doi: 10.7554/eLife.90510 [20] Borsellino P, Krider R I, Chea D, et al. Ketamine and the disinhibition hypothesis: neurotrophic factor-mediated treatment of depression[J]. Pharmaceuticals (Basel), 2023, 16(5): 742. [21] Medeiros G C, Gould T D, Prueitt W L, et al. Blood-based biomarkers of antidepressant response to ketamine and esketamine: a systematic review and meta-analysis[J]. Mol Psychiatry, 2022, 27(9): 3658-3669. [22] Zeng J S, Xie Z Q, 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. [23] Huang E J, Reichardt L F. Trk receptors: roles in neuronal signal transduction[J]. Annu Rev Biochem, 2003, 72: 609-642. [24] Carvalho A L, Caldeira M V, Santos S D, et al. Role of the brain-derived neurotrophic factor at glutamatergic synapses[J]. Br J Pharmacol, 2008, 153(Suppl 1): S310-S324. -
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