PDF(1216 KB)
PDF(1216 KB)
PDF(1216 KB)
对抗大鼠氟西泮抗癫癎耐受性药物及其机制的研究
Drugs and the mechanism for reversing the tolerance of flurazepan in rats
目的：苯二氮卓类药物对各种癫癎均有疗效，长期应用容易产生耐受性，本实验目的是寻找逆转氟西泮抗痫耐受性的药物,探讨神经肽Y(NPY)在此过程中的作用。方法：建立对氟西泮抗痫耐受性的大鼠模型。模型建立后，根据干预药物的种类分为7组：氟西泮耐受组（无药物干预）、硝苯地平组、左乙拉西坦组、托吡酯组、氟马西尼组、L-NAME组、维生素B6组。通过尾静脉注射戊四唑来检测其耐受性,记录其戊四唑致惊厥的潜伏期和惊厥阈值。运用免疫组化法检测海马中NPY表达，通过RT-PCR检测NPY2受体mRNA的改变。结果：与空白对照组相比，氟西泮耐受组的潜伏期缩短、惊厥阈值降低，海马中神经肽Y表达、NPY2受体的mRNA减少。而逆转耐受性后，托吡酯组的潜伏期延长了2倍，惊厥阈值升高了2倍，硝苯地平、左乙拉西坦、氟马西尼组潜伏期延长了1倍，惊厥阈值升高了1倍。氟马西尼组、硝苯地平组和托吡酯组 NPY2受体 mRNA与氟西泮耐受组比较，增加了1~2倍，差异有显著性意义，P＜0.01。结论：硝苯地平、左乙拉西坦、托吡酯、氟马西尼能够逆转氟西泮抗癫癎的耐受性，NPY可能是介导其逆转耐受性因素之一。［中国当代儿科杂志，2010，12（1）：56-61］
OBJECTIVE: Benzodiazepines (BDZ) have many effects on various kinds of epilepsies, but long-term treatment with BDZ often leads to drug tolerance. This study aimed to seek drugs which can reverse the tolerance of flurazepam (FZP), and to explore the role of neuropeptide Y (NPY) in the reversal effect. METHODS: A rat model of anticonvulsant tolerance to FZP was prepared. The rats with FZP tolerance were randomly assigned to seven groups: FZP-tolerance, and nifedipine, levetiracetam, topiramate, flumazenil, L-NAME and pyridoxamine treatment groups. The tolerance to FZP was evaluated through pentylenetetrazol (PTZ) infusion into a tail vein. The latency to onset of clonic seizure and the PTZ threshold were recorded. The mRNA of NPY receptor Y2 in the hippocampus was determined by RT-PCR, and the distribution of NPY in the hippocampus was examined by immunohistochemistry. RESULTS: In comparison with the blank control group, the average latency to the onset of clonic seizure was shortened, the average PTZ threshold decreased and the expression of NYT and NPY receptor Y2 mRNA decreased significantly in the FZP-tolerance group (P<0.01). In comparison with the FZP-tolerance group, the average latency to onset of clonic seizure was prolonged by 2 times and the average PTZ threshold doubled in the topiramate treatment group. The average latency to onset of clonic seizure was prolonged by 1 time and the average PTZ threshold increased 1 time in the nifedipine, the levetiracetam and the flumazenil treatment groups. The mRNA expression of NPY receptor Y2 increased by 1 or 2 times in the flumazenil, the nifedipine and the topiramate treatment groups when compared with the FZP-tolerance group. CONCLUSIONS: Nifedipine, levetiracetam, topiramate and flumazenil can reverse the anticonvulsant tolerance to flurazepam. NPY may play a role in mediating the reversal effect.［Chin J Contemp Pediatr, 2010, 12 (1):56-61］
氟西泮 / 耐受性 / 神经肽Y / 神经肽Y2受体 / 大鼠
Flurazepam / Tolerance / Neuropeptide Y / Neuropeptide Y2 receptor / Rats
［1］王晓东,王丽,陶成.氯硝西泮抗大鼠听源性惊厥耐受性的受体学基础［J］.中国药理学通报, 1996, 12(3):224-226.
［2］Xiang K, Tietz EI.Benzodiazepine-induced hippocampal CA1 neuron alpha-amino-3-hydroxy-5-methylisoxasole-4-propionic acid (AMPA) receptor plasticity linked to severity of withdrawal anxiety: differential role of voltage-gated calcium channels and N-methyl-D-aspartic acid receptors［J］. Behav Pharmacol, 2007, 18(5-6):447-460.
［3］张礼萍,王丽.大鼠对氟西泮抗痫耐受和依赖时海马中一氧化氮合酶的变化［J］.中国药理学通报, 2005, 21(8):927-930.
［4］Xiang K, Earl DE, Davis KM. Chronic benzodiazepine administration potentiates high voltage-activated calcium currents in hippocampal CA1 neurons［J］. Pharmacol Exp Ther, 2008, 327(3):872-883.
［5］Blomqvist AG, Herzog H. Y-receptor subtypes—how many more?［J］.Trends Neurosci, 1997, 20(7):294-298.
［6］Ozawa S, Kamiya H, Tsuzuki K. Glutamate receptors in the mammalian central nervous system［J］. Prog Neurobiol, 1998, 54(5):581-618.
［7］Silva AP, Carvalho AP, Carvalho CM and Malva JO. Functional interaction between neuropeptide Y receptors and modulation of calcium channels in the rat hippocampus［J］. Neuropharmacology, 2003, 42(2):282-292.
［8］Zhang LP, Wang L. Changes of brain neuropeptide Y and its receptors in rats with flurazepam tolerance and dependence1 ［J］. Acta Pharmacologica Sinica, 2005, 26(11):1290-1296.
［9］Rosenberg HC. Differential expression of benzodiazepine anticonvulsant cross-tolerance according to time following flurazepam or diazepam treatment［J］. Pharmacol Biochem Behav, 1995, 51(2):363-368.
［10］Xiang K, Tietz EI. Chronic benzodiazepine-induced reduction in gabaa receptor mediated synaptic currents in hippocampal pyramidal neurons prevented by prior nimodipine injection［J］. Neuroscience, 2008, 157(1):153-163.
［11］Kaminski RM, Gillard M, Leclercq K, Hanon E, Lorent G, Dassesse D, et al. Proepileptic phenotype of SV2A-deficient mice is associated with reduced anticonvulsant efficacy of levetiracetam［J］. Epilepsia, 2009, 50(7):1729-1740.
［12］Husum H, Bolwig TG, Sánchez C, Mathé AA, Hansen SL.Levetiracetam prevents changes in levels of brain-derived neurotrophic factor and neuropeptide Y mRNA and of Y1-and Y5-like receptors in the hippocampus of rats undergoing amygdala kindling: implications for antiepileptogenic and mood-stabilizing properties［J］. Epilepsy Behav, 2004, 5(1):204-215.
［13］Lynch BA, Lambeng N, Nocka K, Kensel-Hammes P, Bajjalieh SM, Matagne A,et al. The synaptic vesicle protein SV2A is the binding site for the antiepileptic drug levetiracetam［J］. Proc Natl Acad Sci U S A, 2004, 101(26):9861-9866.
［14］Pisani A, Bonsi P, Martella G, De Persis C, Costa C, Pisani F,et al. Intracellular calcium increase in epileptiform activity: modulation by levetiracetam and lamotrigine［J］. Epilepsia, 2004, 45(7):719-728.
［15］Ueda Y, Doi T, Nagatomo K, Tokumaru J, Takaki M, Willmore LJ.Effect of levetiracetam on molecular regulation of hippocampal glutamate and GABA transporters in rats with chronic seizures induced by amygdalar FeCl3 injection［J］. Brain Re, 2007, 1151:55-61.
［16］Meldrum B. Action of established and novel anticonvulsant drugs on the basic mechanisms of epilepsy［J］. Epilepsy, 1996, 11(9):67-77.
［17］Russo E, Constanti A, Ferreri G, Citraro R, De Sarro G. Nifedipine affects the anticonvulsant activity of topiramate in various animal models of epilepsy［J］. Neuropharmacology, 2004, 46（6）:865-878.
［18］朱遂强，杨美兰，陈景刚，李卫华，苟玉兰，阮旭中.托吡酯对戊四氮活化培养海马神经元的作用［J］.华中科技大学学报 (医学版), 2004, 33(3):350-352.
［19］郭文,王丽.氟马西尼逆转惊厥易感大鼠氟西泮耐受性的分子机制［J］.北京大学学报, 2001, 33（5）:452-456.
［20］Hertz L, Zhao Z, Chen Y. The astrocytic GABA(A)/benzodiazepine-like receptor: the Joker receptor for benzodiazepine-mimetic drugs? ［J］.Recent Pat CNS Drug Discov, 2006, 1(1):93-103.
［21］Flaishon R, Weinbroum AA, Veenman L, Leschiner S, Rudick V, Gavish M. Flumazenil attenuates development of tolerance to diazepam after chronic treatment of mice with either isoflurane or diazepam ［J］. Anesth Analg, 2003, 97(4):1046-1052.
［22］Bayoumi R, Kirwan JR, Smith WR. Some effects of dietary vitamin B6 deficiency and 4-deoxypiridoxine on γ-aminobutyric acid metabolism in rat brain［J］. J Neurochemistry, 1972, 19(3):569-576.
［23］Patrylo PR, van den Pol AN, Spencer DD, Williamson A. NPY inhibits glutamaterigic exicitation in the epileptic human dentate gyrus［J］.J Neurophysiol, 1999, 82(1):478-483.
［24］Silva AP, Carvalho AP, Carvalho CM, Malva JO.Functional interaction between neuropeptide Y receptors and modulation of calcium channels in the rat hippocampus［J］. Neuropharmacology, 2003, 44(2):282-292.
［25］Silva AP, Carvalho AP, Carvalho CM, Malva JO. Modulation of intracellular calcium changes and glutamate release by neuropeptide Y1 and Y2 receptors in the rat hippocampus: differential effects in CA1, CA3 and dentate gyrus［J］.J Neurochem, 2001, 79(2):286-296.
［26］Colmers WF, Klapstein GJ, Fournier A, StPierre S, Treherne KA. Presynaptic inhibition by neuropetide Y in rat hippocampal slice in vitro is mediated by a Y2 receptor［J］. Br J Pharmacol, 1991, 102(1):41-44.
［27］Foti S, Haberman RP, Samulski RJ, McCown TJ. Adeno-associated virus-mediated expression and constitutive secretion of NPY or NPY13-36 suppresses seizure activity in vivo［J］. Gene Therapy, 2007, 14(21):1534-1536.
