利鲁唑上调EAATs产生对MPTP诱导的帕金森模型小鼠的保护作用
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摘要
目的探讨MPTP对EAATs的影响及利鲁唑对其诱发帕金森小鼠模型的保护作用。方法选取42只小鼠,随机分为3组:对照组(生理盐水组)、MPTP组(PD模型组)和MPTP+利鲁唑组(利鲁唑治疗组)。采用小鼠转棒实验和疲劳仪实验评价三组小鼠行为学差异;南京建成试剂盒检测纹状体谷氨酸(Glutamate,Glu)含量;Western blot和RT-PCR方法检测小鼠纹状体EAAT1及EAAT2的表达。结果与对照组比较,小鼠转棒潜伏期、疲劳耐力时间及跑步距离均明显减少(P<0.01),Glu含量明显增加(P<0.01),EAAT1及EAAT2 mRNA表达和蛋白水平显著下降。与MPTP组比较,利鲁唑治疗组小鼠转棒潜伏期、疲劳耐力时间及跑步距离均明显增加(P<0.01),Glu含量明显下降(P<0.01),EAAT1及EAAT2 mRNA表达和蛋白水平显著升高。结论利鲁唑可通过上调EAAT1和EAAT2对MPTP诱导的帕金森模型小鼠的运动功能起到保护作用。
Objective To observe the effect of MPTP on EAATs and the protection effect of riluzole on MPTP-induced Parkinson′s disease in mice model.Methods Totally 42 healthy mice were randomly divided into 3 groups: control group(NS group),MPTP group(PD model group)and MPTP+riluzole group(treatment group).Rotarod experiment and fatigue tester system were used to evaluate the behavioral differences among the animals in 3 groups;Nanjing Jiancheng kit was used to test the levels of glutamate(Glu)in striatum;Western blot and RT-PCR were used to detect the levels of EAAT1 and EAAT2 in striatum.Results Compared with control group,the rotarod latency,fatigue endurance time,running distance,mRNA and protein of EAAT1 and EAAT2 were decreased in MPTP group significantly,while the levels of Glu increased.Compared with MPTP group,all these indexes except Glu levels in MPTP+riluzole group were increased,while Glu level was decreased.Conclusion Riluzole can up-regulate the EAAT1 and EAAT2 to exert protective effects on MPTP-induced Parkinson′s disease mice model.
Objective To observe the effect of MPTP on EAATs and the protection effect of riluzole on MPTP-induced Parkinson′s disease in mice model.Methods Totally 42 healthy mice were randomly divided into 3 groups: control group(NS group),MPTP group(PD model group)and MPTP+riluzole group(treatment group).Rotarod experiment and fatigue tester system were used to evaluate the behavioral differences among the animals in 3 groups;Nanjing Jiancheng kit was used to test the levels of glutamate(Glu)in striatum;Western blot and RT-PCR were used to detect the levels of EAAT1 and EAAT2 in striatum.Results Compared with control group,the rotarod latency,fatigue endurance time,running distance,mRNA and protein of EAAT1 and EAAT2 were decreased in MPTP group significantly,while the levels of Glu increased.Compared with MPTP group,all these indexes except Glu levels in MPTP+riluzole group were increased,while Glu level was decreased.Conclusion Riluzole can up-regulate the EAAT1 and EAAT2 to exert protective effects on MPTP-induced Parkinson′s disease mice model.
引文
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[13] Ngm O,Kll M,Munster EM,et al.The effect of a structured medication review on quality of life in Parkinson′s disease:the study protocol[J].Contemp Clin Trials Commun,2019,13:100308.
[14] Assous M,Had-Aissouni L,Gubellini P,et al.Progressive Parkinsonism by acute dysfunction of excitatory amino acid transporters in the rat substantia nigra[J].Neurobiol Dis,2014,65:69-81.
[15] Kong Q,Chang LC,Takahashi K,et al.Small-molecule activator of glutamate transporter EAAT2 translation provides neuroprotection[J].J Clin Invest,2014,124(3):1255-1267.
[16] Rothstein JD,Patel S,Regan MR,et al.Beta-lactam antibiotics offer neuroprotection by increasing glutamate transporter expression[J].Nature,2005,433(7021):73-77.
[17] Ruzza P,Siligardi G,Hussain R,et al.Ceftriaxone blocks the polymerization of α-synuclein and exerts neuroprotective effects in vitro[J].ACS Chem Neurosci,2014,5(1):30-38.
[18] Petr GT,Sun Y,Frederick NM,et al.Conditional deletion of the glutamate transporter GLT-1 reveals that astrocytic GLT-1 protects against fatal epilepsy while neuronal GLT-1 contributes significantly to glutamate uptake into synaptosomes[J].J Neurosci,2015,35(13):5187-5201.
[19] Barnéoud P,Mazadier M,Miquet JM,et al.Neuroprotective effects of riluzole on a model of Parkinson′s disease in the rat[J].Neuroscience,1996,74(4):971-983.
[20] Boireau A,Dubedat P,Bordier F,et al.The protective effect of riluzole in the MPTP model of Parkinson′s disease in mice is not due to a decrease in MPP(+)accumulation[J].Neuropharmacology,2000,39(6):1016-1020.
[21] Obinu MC,Reibaud M,Blanchard V,et al.Neuroprotective effect of riluzole in a primate model of Parkinson′s disease:behavioral and histological evidence[J].Mov Disord,2002,17(1):13-19.
[2] Goldman SM,Tanne CM,Korell M,et al.Environmental epidemiology of Parkinson′s disease[C].北京,2007.
[3] 林剑霞,潘瑶,李妍.帕金森病的研究进展[J].吉林医药学院学报,2015,36(2):144-147.
[4] Van Laar VS,Roy N,Liu A,et al.Glutamate excitotoxicity in neurons triggers mitochondrial and endoplasmic reticulum accumulation of Parkin,and,in the presence of N-acetyl cysteine,mitophagy[J].Neurobiol Dis,2015,74:180-193.
[5] Zhang Y,He X,Meng X,et al.Regulation of glutamate transporter trafficking by Nedd4-2 in a Parkinson′s disease model[J].Cell Death Dis,2017,8(2):e2574.
[6] Chung EK,Chen LW,Chan YS,et al.Downregulation of glial glutamate transporters after dopamine denervation in the striatum of 6-hydroxydopamine-lesioned rats[J].J Comp Neurol,2008,511(4):421-437.
[7] Mitsumoto H.A strategy to develop effective ALS therapy[J].Brain Nerve,2007,59(4):383-391.
[8] Irifune M,Kikuchi N,Saida T,et al.Riluzole,a glutamate release inhibitor,induces loss of righting reflex,antinociception,and immobility in response to noxious stimulation in mice[J].Anesth Analg,2007,104(6):1415-1421.
[9] Fumagalli E,Funicello M,Rauen T,et al.Riluzole enhances the activity of glutamate transporters GLAST,GLT1 and EAAC1[J].Eur J Pharmacol,2008,578(2-3):171-176.
[10] Jankovic J,Hunter C.A double-blind,placebo-controlled and longitudinal study of riluzole in early Parkinson′s disease[J].Parkinsonism Relat Disord,2002,8(4):271-276.
[11] Aoyama K,Matsumura N,Watabe M,et al.Oxidative stress on EAAC1 is involved in MPTP-induced glutathione depletion and motor dysfunction[J].Eur J Neurosci,2008,27(1):20-30.
[12] Wu JY,Niu FN,Huang R,et al.Enhancement of glutamate uptake in 1-methyl-4-phenylpyridinium-treated astrocytes by trichostatin A[J].Neuroreport,2008,19(12):1209-1212.
[13] Ngm O,Kll M,Munster EM,et al.The effect of a structured medication review on quality of life in Parkinson′s disease:the study protocol[J].Contemp Clin Trials Commun,2019,13:100308.
[14] Assous M,Had-Aissouni L,Gubellini P,et al.Progressive Parkinsonism by acute dysfunction of excitatory amino acid transporters in the rat substantia nigra[J].Neurobiol Dis,2014,65:69-81.
[15] Kong Q,Chang LC,Takahashi K,et al.Small-molecule activator of glutamate transporter EAAT2 translation provides neuroprotection[J].J Clin Invest,2014,124(3):1255-1267.
[16] Rothstein JD,Patel S,Regan MR,et al.Beta-lactam antibiotics offer neuroprotection by increasing glutamate transporter expression[J].Nature,2005,433(7021):73-77.
[17] Ruzza P,Siligardi G,Hussain R,et al.Ceftriaxone blocks the polymerization of α-synuclein and exerts neuroprotective effects in vitro[J].ACS Chem Neurosci,2014,5(1):30-38.
[18] Petr GT,Sun Y,Frederick NM,et al.Conditional deletion of the glutamate transporter GLT-1 reveals that astrocytic GLT-1 protects against fatal epilepsy while neuronal GLT-1 contributes significantly to glutamate uptake into synaptosomes[J].J Neurosci,2015,35(13):5187-5201.
[19] Barnéoud P,Mazadier M,Miquet JM,et al.Neuroprotective effects of riluzole on a model of Parkinson′s disease in the rat[J].Neuroscience,1996,74(4):971-983.
[20] Boireau A,Dubedat P,Bordier F,et al.The protective effect of riluzole in the MPTP model of Parkinson′s disease in mice is not due to a decrease in MPP(+)accumulation[J].Neuropharmacology,2000,39(6):1016-1020.
[21] Obinu MC,Reibaud M,Blanchard V,et al.Neuroprotective effect of riluzole in a primate model of Parkinson′s disease:behavioral and histological evidence[J].Mov Disord,2002,17(1):13-19.