原肌球蛋白相关激酶B受体及其信号通路在低频重复经颅磁刺激治疗癫痫大鼠中的作用
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摘要
目的探索原肌球蛋白相关激酶B受体(TrkB)表达和TrkB信号通路活性在低频重复经颅磁刺激(rTMS)治疗癫痫大鼠过程中的变化以及作用机制。方法构建匹罗卡品诱导的癫痫大鼠模型,采用低频rTMS(1 Hz)干预癫痫大鼠模型后,Western Blot检测TrkB受体全长型和截短型蛋白表达以及全长型TrkB信号通路磷酸化水平的变化情况;膜片钳检测神经元自发放电频率的变化情况。结果经匹罗卡品诱导构建的癫痫大鼠模型,其神经元放电频率较对照组明显增加(P <0.05)。与对照组相比,癫痫模型中全长型TrkB受体表达降低(P <0.05),而截短型表达升高(P <0.05),此外全长型TrkB受体磷酸化程度降低(P <0.05)。低频rTMS干预癫痫模型后,与癫痫组相比,全长型TrkB受体表达上调(P <0.05),截短型TrkB受体表达下调(P <0.05),全长型TrkB受体磷酸化水平上调并激活TrkB信号通路(P <0.05)。最终,癫痫模型经低频rTMS干预后,其自发放电频率明显降低(P <0.05)。结论低频rTMS可能是通过上调癫痫模型中低表达的全长型TrkB受体,下调高表达的截短型TrkB受体,并激活全长型TrkB信号通路,最终发挥其抗癫痫作用。
Objective To explore the expression changes and effects of tropomyosin-related kinase B receptor(TrkB)and TrkB signalingin the treatment of epileptic rats by low frequency repetitive transcranial magnetic stimulation(rTMS). Methods Pilocarpine-induced status epilepticus rat model were constructed. After the intervention of low frequency rTMS(1 Hz)in epileptic rats,the changes in the expression of full-length and truncated TrkB receptor and the phosphorylation of the full-length TrkB signaling were detected by Western blot,then the changes in spontaneous discharge frequency of neurons were detected by patch clamp. Results The discharge frequency of neurons in pilocarpine-induced status epilepticus rat model was significantly higher than that of the control group(P < 0.05). Compared with the control group,the expression of the full-length TrkB was decreased(P < 0.05)and the truncated TrkB was increased(P < 0.05)in the epileptic model. Furthermore,the phosphorylation of full-length TrkB was decreased(P < 0.05). After the intervention of low frequency rTMS in epileptic rats,compared with the epileptic group,the full-length TrkB was upregulated(P < 0.05)and truncated TrkB was downregulated(P < 0.05),and the TrkB signaling was activated by the increase of the phosphorylation of full-length TrkB receptor(P < 0.05). Finally,the discharge frequency of neurons in epileptic models was decreased significantly after low frequency rTMS(P < 0.05). Conclusion Low frequency rTMS might upregulate the expression of full-length TrkB receptor,downregulate the truncated TrkB receptor and activate the full length TrkB signaling in the epileptic model to exertanti-epileptic effect.
Objective To explore the expression changes and effects of tropomyosin-related kinase B receptor(TrkB)and TrkB signalingin the treatment of epileptic rats by low frequency repetitive transcranial magnetic stimulation(rTMS). Methods Pilocarpine-induced status epilepticus rat model were constructed. After the intervention of low frequency rTMS(1 Hz)in epileptic rats,the changes in the expression of full-length and truncated TrkB receptor and the phosphorylation of the full-length TrkB signaling were detected by Western blot,then the changes in spontaneous discharge frequency of neurons were detected by patch clamp. Results The discharge frequency of neurons in pilocarpine-induced status epilepticus rat model was significantly higher than that of the control group(P < 0.05). Compared with the control group,the expression of the full-length TrkB was decreased(P < 0.05)and the truncated TrkB was increased(P < 0.05)in the epileptic model. Furthermore,the phosphorylation of full-length TrkB was decreased(P < 0.05). After the intervention of low frequency rTMS in epileptic rats,compared with the epileptic group,the full-length TrkB was upregulated(P < 0.05)and truncated TrkB was downregulated(P < 0.05),and the TrkB signaling was activated by the increase of the phosphorylation of full-length TrkB receptor(P < 0.05). Finally,the discharge frequency of neurons in epileptic models was decreased significantly after low frequency rTMS(P < 0.05). Conclusion Low frequency rTMS might upregulate the expression of full-length TrkB receptor,downregulate the truncated TrkB receptor and activate the full length TrkB signaling in the epileptic model to exertanti-epileptic effect.
引文
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[10] SUN W,MAO W,MENG X,et al. Low-frequency repetitive transcranial magnetic stimulation for the treatment of refractory partial epilepsy:a controlled clinical study[J]. Epilepsia,2012,53(10):1782-1789.
[11] COOPER Y A,PIANKA S T,ALOTAIBI N M,et al. Repetitive transcranial magnetic stimulation for the treatment of drugresistant epilepsy:A systematic review and individual participant data meta-analysis of real-world evidence[J]. Epilepsia Open,2018,3(1):55-65.
[12] KOTLOSKI R,MCNAMARA J O. Reduction of TrkB expression de novo in the adult mouse impairs epileptogenesis in the kindling model[J]. Hippocampus,2010,20(6):713-723.
[13] UNSAIN N,NUNEZ N,ANASTASIA A,et al. Status epilepticus induces a TrkB to p75 neurotrophin receptor switch and increases brain-derived neurotrophic factor interaction with p75neurotrophin receptor:an initial event in neuronal injury induction[J]. Neuroscience,2008,154(3):978-993.
[14] XIE W,SONG Y J,LI D,et al. The suppression of epileptiform discharges in cultured hippocampal neurons is regulated via alterations in full-length tropomyosin-related kinase type B receptors signalling activity[J]. Eur J Neurosci,2014,40(3):2564-2575.
[15] XIANG L,REN Y,CAI H,et al. MicroRNA-132 aggravates epileptiform discharges via suppression of BDNF/TrkB signaling in cultured hippocampal neurons[J]. Brain Res,2015,1622:484-495.
[2] KISTSEN V,EVSTIGNEEV V,DUBOVIK B,et al. The effects of repetitive transcranial magnetic stimulation on picrotoxin-induced convulsions in mice[J]. Adv Clin Exp Med,2016,25(2):317-325.
[3]崔桂雪,闫海亮,于毅,等.重复经颅磁刺激对阿尔茨海默病大鼠脑电LZ复杂度和脑白质中碱性髓鞘蛋白分布的影响[J].实用医学杂志,2017,33(15):2468-2473.
[4]魏宇佳,石先俊,刘立红,等.结合导航系统的经颅磁刺激在功能区癫痫术前评估中的应用[J].中华神经医学杂志,2017,16(10):978-983.
[5]任萍,秦幸娜,李小俚.经颅磁刺激干预自闭症的理论基础和应用[J].中华神经医学杂志,2017,16(6):643-648.
[6] SANCHEZ-ESCANDON O,ARANA-LECHUGA Y,TERANPEREZ G,et al. Effect of low-frequency repetitive transcranial magnetic stimulation on sleep pattern and quality of life in patients with focal epilepsy[J]. Sleep Med,2016,20:37-40.
[7] HELGAGER J,LIU G,MCNAMARA J O. The cellular and synaptic location of activated TrkB in mouse hippocampus during limbic epileptogenesis[J]. J Comp Neurol,2013,521(3):499-521,Spc1.
[8] HEINRICH C,LAHTEINEN S,SUZUKI F,et al. Increase in BDNF-mediated TrkB signaling promotes epileptogenesis in a mouse model of mesial temporal lobe epilepsy[J]. Neurobiol Dis,2011,42(1):35-47.
[9] MA J,ZHANG Z,SU Y,et al. Magnetic stimulation modulates structural synaptic plasticity and regulates BDNF-TrkB signal pathway in cultured hippocampal neurons[J]. Neurochem Int,2013,62(1):84-91.
[10] SUN W,MAO W,MENG X,et al. Low-frequency repetitive transcranial magnetic stimulation for the treatment of refractory partial epilepsy:a controlled clinical study[J]. Epilepsia,2012,53(10):1782-1789.
[11] COOPER Y A,PIANKA S T,ALOTAIBI N M,et al. Repetitive transcranial magnetic stimulation for the treatment of drugresistant epilepsy:A systematic review and individual participant data meta-analysis of real-world evidence[J]. Epilepsia Open,2018,3(1):55-65.
[12] KOTLOSKI R,MCNAMARA J O. Reduction of TrkB expression de novo in the adult mouse impairs epileptogenesis in the kindling model[J]. Hippocampus,2010,20(6):713-723.
[13] UNSAIN N,NUNEZ N,ANASTASIA A,et al. Status epilepticus induces a TrkB to p75 neurotrophin receptor switch and increases brain-derived neurotrophic factor interaction with p75neurotrophin receptor:an initial event in neuronal injury induction[J]. Neuroscience,2008,154(3):978-993.
[14] XIE W,SONG Y J,LI D,et al. The suppression of epileptiform discharges in cultured hippocampal neurons is regulated via alterations in full-length tropomyosin-related kinase type B receptors signalling activity[J]. Eur J Neurosci,2014,40(3):2564-2575.
[15] XIANG L,REN Y,CAI H,et al. MicroRNA-132 aggravates epileptiform discharges via suppression of BDNF/TrkB signaling in cultured hippocampal neurons[J]. Brain Res,2015,1622:484-495.