美金刚通过调节ERK、CREB信号通路及突触可塑性保护脑缺血再灌注小鼠的作用研究
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摘要
目的研究美金刚对脑缺血再灌注小鼠模型ERK、CREB信号通路及突触可塑性的作用机制。方法60只小鼠随机分为假手术组(Sham),模型组(MCAO),美金刚20 mg/kg组(M20),美金刚4 mg/kg组(M4),生理盐水组(NS),除假手术组外,其余4组均使用线栓法建立小鼠大脑中动脉梗死模型,观察小鼠的体重变化及神经功能缺损程度,甲酚紫染色观察小鼠脑萎缩体积,粘附物去除实验观察缺血小鼠的运动及感觉功能变化,Morris水迷宫检测小鼠的认知记忆功能,Western Blot检测ERK1/2、p-ERK1/2、CREB、p-CREB、PSD-95、Synaptophysin等蛋白的表达情况。结果与模型组(MCAO)、美金刚4 mg/kg组(M4)及生理盐水组(NS)相比,美金刚20 mg/kg组的小鼠体重下降程度减小,神经功能缺损评分下降,脑萎缩体积减轻,小鼠感觉运动功能出现改善,水迷宫实验结果提示学习记忆损伤程度减轻,p-ERK1/2、p-CREB、PSD-95、synaptophysin等蛋白表达升高。结论小鼠脑缺血后持续给予美金刚20 mg/kg治疗,可以提高小鼠的神经功能恢复,改善小鼠的学习记忆功能,改善了大脑的突触可塑性。
Objective To investigate the mechanism of memantine on ERK/CREB signaling and synaptic plasticity in a mouse model of cerebral ischemia-reperfusion. Methods Sixty mice were randomly divided into a sham operation group( Sham),middle cerebral artery occlusion model group( MCAO),20 mg/kg memantine group( M20),4 mg/kg memantine group( M4),and saline group( NS). With the exception of the Sham group,ischemia-reperfusion was established in groups using a suture to achieve MCAO. Changes in body weight and degree of neurological deficits were observed in the mice. Brain atrophy volume was measured using cresyl violet staining,while sensorimotor function was observed by an adhesive removal test and cognitive function was detected by Morris water maze. Expression of ERK1/2,phosphorylated ERK1/2( p-ERK1/2),CREB,p-CREB,postsynaptic density protein 95( PSD-95) and synaptophysin were detected by western blot assay. Results Compared with MCAO,M4,and NS groups,the M20 group exhibited reduced weight loss,decreased neurological severity scores,decreased volume of brain atrophy,and improved sensorimotor function. In addition,the results of water maze testing showed that the degree of learning and memory impairment was reduced in the M20 group. Expression of p-ERK1/2,p-CREB,PSD-95,and synaptophysin proteins were increased in the M20 group compared with other groups. Conclusions Continuous treatment with memantine( 20 mg/kg) after cerebral ischemia in mice can improve the recovery of neurological function,ameliorate deficits in learning and memory,and improve synaptic plasticity of the brain.
Objective To investigate the mechanism of memantine on ERK/CREB signaling and synaptic plasticity in a mouse model of cerebral ischemia-reperfusion. Methods Sixty mice were randomly divided into a sham operation group( Sham),middle cerebral artery occlusion model group( MCAO),20 mg/kg memantine group( M20),4 mg/kg memantine group( M4),and saline group( NS). With the exception of the Sham group,ischemia-reperfusion was established in groups using a suture to achieve MCAO. Changes in body weight and degree of neurological deficits were observed in the mice. Brain atrophy volume was measured using cresyl violet staining,while sensorimotor function was observed by an adhesive removal test and cognitive function was detected by Morris water maze. Expression of ERK1/2,phosphorylated ERK1/2( p-ERK1/2),CREB,p-CREB,postsynaptic density protein 95( PSD-95) and synaptophysin were detected by western blot assay. Results Compared with MCAO,M4,and NS groups,the M20 group exhibited reduced weight loss,decreased neurological severity scores,decreased volume of brain atrophy,and improved sensorimotor function. In addition,the results of water maze testing showed that the degree of learning and memory impairment was reduced in the M20 group. Expression of p-ERK1/2,p-CREB,PSD-95,and synaptophysin proteins were increased in the M20 group compared with other groups. Conclusions Continuous treatment with memantine( 20 mg/kg) after cerebral ischemia in mice can improve the recovery of neurological function,ameliorate deficits in learning and memory,and improve synaptic plasticity of the brain.
引文
[1] Prabhakaran S,Ruff I,Bernstein RA. Acute stroke intervention:a systematic review[J]. JAMA,2015,313(14):1451-1462.
[2] Iacobucci GJ, Popescu GK. NMDA receptors:linking physiological output to biophysical operation[J]. Nat Rev Neurosci,2017,18(4):236-249.
[3] Black SA,Stys PK,Zamponi GW,et al. Cellular prion protein and NMDA receptor modulation:protecting against excitotoxicity[J]. Front Cell Dev Biol,2014,2:45.
[4] López-Valdés HE,Clarkson AN,Ao Y, et al. Memantine enhances recovery from stroke[J]. Stroke,2014,45(7):2093-2100.
[5] Li Y,Li X,Guo C,et al. Long-term neurocognitive dysfunction in offspring via NGF/ERK/CREB signaling pathway caused by ketamine exposure during the second trimester of pregnancy in rats[J]. Oncotarget,2017,8(19):30956-30970.
[6] Culmsee C,Junker V,Kremers W,et al. Combination therapy in ischemic stroke:synergistic neuroprotective effects of memantine and clenbuterol[J]. Stroke,2004,35(5):1197-1202.
[7] Wang YC,Sanchez-Mendoza EH,Doeppner TR,et al. Postacute delivery of memantine promotes post-ischemic neurological recovery,peri-infarct tissue remodeling,and contralesional brain plasticity[J]. J Cereb Blood Flow Metab,2017,37(3):980-993.
[8] Cai H,Ma Y,Jiang L,et al. Hypoxia response elementregulated MMP-9 promotes neurological recovery via glial scar degradation and angiogenesis in delayed stroke[J]. Mol Ther,2017,25(6):1448-1459.
[9] Choi I A,Lee C S,Kim H Y,et al. Effect of inhibition of DNA methylation combined with task-specific training on chronic stroke recovery[J]. Int J Mol Sci,2018,19(7):pii:E2019.
[10] Ma S,Wang J,Wang Y,et al. Diabetes mellitus impairs white matter repair and long-term functional deficits after cerebral ischemia[J]. Stroke,2018,49(10):2453-2463.
[11] Wikinski SI,Acosta GB. Role of excitatory amino acids in neuropathology[J]. Medicina(B Aires),1995,55(4):355-365.
[12] Chen ZZ, Yang DD, Zhao Z, et al. Memantine mediates neuroprotection via regulating neurovascular unit in a mouse model of focal cerebral ischemia[J]. Life Sci,2016,150:8-14.
[13] Liang HM,Geng LJ,Shi XY,et al. By up-regulating mu-and delta-opioid receptors, neuron-restrictive silencer factor knockdown promotes neurological recovery after ischemia[J].Oncotarget,2017,8(60):101012-101025.
[14] Zhang X,Zhang Q,Tu J,et al. Prosurvival NMDA 2A receptor signaling mediates postconditioning neuroprotection in the hippocampus[J]. Hippocampus,2015,25(3):286-296.
[15] Chen L,Miyamoto Y,Furuya K,et al. PREGS induces LTP in the hippocampal dentate gyrus of adult rats via the tyrosine phosphorylation of NR2B coupled to ERK/CREB[corrected]signaling[J]. J Neurophysiol,2007,98(3):1538-1548.
[16] Coley AA,Gao WJ. PSD95:A synaptic protein implicated in schizophrenia or autism?[J]. Prog Neuropsychopharmacol Biol Psychiatry,2018,82:187-194.
[17] Osimo EF,Beck K,Reis Marques T,et al. Synaptic loss in schizophrenia:a meta-analysis and systematic review of synaptic protein and mRNA measures[J]. Mol Psychiatry,2019,24(4):549-561.
[18] Wi S,Yu JH,Kim M,et al. In vivo expression of reprogramming factors increases hippocampal neurogenesis and synaptic plasticity in chronic hypoxic-ischemic brain injury[J]. Neural Plast,2016,2016:2580837.
[19] Qu H,Zhao M,Zhao S,et al. Forced limb-use enhances brain plasticity through the c AMP/PKA/CREB signal transduction pathway after stroke in adult rats[J]. Restor Neurol Neurosci,2014,32(5):597-609.
[2] Iacobucci GJ, Popescu GK. NMDA receptors:linking physiological output to biophysical operation[J]. Nat Rev Neurosci,2017,18(4):236-249.
[3] Black SA,Stys PK,Zamponi GW,et al. Cellular prion protein and NMDA receptor modulation:protecting against excitotoxicity[J]. Front Cell Dev Biol,2014,2:45.
[4] López-Valdés HE,Clarkson AN,Ao Y, et al. Memantine enhances recovery from stroke[J]. Stroke,2014,45(7):2093-2100.
[5] Li Y,Li X,Guo C,et al. Long-term neurocognitive dysfunction in offspring via NGF/ERK/CREB signaling pathway caused by ketamine exposure during the second trimester of pregnancy in rats[J]. Oncotarget,2017,8(19):30956-30970.
[6] Culmsee C,Junker V,Kremers W,et al. Combination therapy in ischemic stroke:synergistic neuroprotective effects of memantine and clenbuterol[J]. Stroke,2004,35(5):1197-1202.
[7] Wang YC,Sanchez-Mendoza EH,Doeppner TR,et al. Postacute delivery of memantine promotes post-ischemic neurological recovery,peri-infarct tissue remodeling,and contralesional brain plasticity[J]. J Cereb Blood Flow Metab,2017,37(3):980-993.
[8] Cai H,Ma Y,Jiang L,et al. Hypoxia response elementregulated MMP-9 promotes neurological recovery via glial scar degradation and angiogenesis in delayed stroke[J]. Mol Ther,2017,25(6):1448-1459.
[9] Choi I A,Lee C S,Kim H Y,et al. Effect of inhibition of DNA methylation combined with task-specific training on chronic stroke recovery[J]. Int J Mol Sci,2018,19(7):pii:E2019.
[10] Ma S,Wang J,Wang Y,et al. Diabetes mellitus impairs white matter repair and long-term functional deficits after cerebral ischemia[J]. Stroke,2018,49(10):2453-2463.
[11] Wikinski SI,Acosta GB. Role of excitatory amino acids in neuropathology[J]. Medicina(B Aires),1995,55(4):355-365.
[12] Chen ZZ, Yang DD, Zhao Z, et al. Memantine mediates neuroprotection via regulating neurovascular unit in a mouse model of focal cerebral ischemia[J]. Life Sci,2016,150:8-14.
[13] Liang HM,Geng LJ,Shi XY,et al. By up-regulating mu-and delta-opioid receptors, neuron-restrictive silencer factor knockdown promotes neurological recovery after ischemia[J].Oncotarget,2017,8(60):101012-101025.
[14] Zhang X,Zhang Q,Tu J,et al. Prosurvival NMDA 2A receptor signaling mediates postconditioning neuroprotection in the hippocampus[J]. Hippocampus,2015,25(3):286-296.
[15] Chen L,Miyamoto Y,Furuya K,et al. PREGS induces LTP in the hippocampal dentate gyrus of adult rats via the tyrosine phosphorylation of NR2B coupled to ERK/CREB[corrected]signaling[J]. J Neurophysiol,2007,98(3):1538-1548.
[16] Coley AA,Gao WJ. PSD95:A synaptic protein implicated in schizophrenia or autism?[J]. Prog Neuropsychopharmacol Biol Psychiatry,2018,82:187-194.
[17] Osimo EF,Beck K,Reis Marques T,et al. Synaptic loss in schizophrenia:a meta-analysis and systematic review of synaptic protein and mRNA measures[J]. Mol Psychiatry,2019,24(4):549-561.
[18] Wi S,Yu JH,Kim M,et al. In vivo expression of reprogramming factors increases hippocampal neurogenesis and synaptic plasticity in chronic hypoxic-ischemic brain injury[J]. Neural Plast,2016,2016:2580837.
[19] Qu H,Zhao M,Zhao S,et al. Forced limb-use enhances brain plasticity through the c AMP/PKA/CREB signal transduction pathway after stroke in adult rats[J]. Restor Neurol Neurosci,2014,32(5):597-609.
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