切换至 "中华医学电子期刊资源库"

中华妇幼临床医学杂志(电子版) ›› 2009, Vol. 05 ›› Issue (01) : 13 -17. doi: 10.3877/cma.j.issn.1673-5250.2009.01.106

论著

戊四氮致癫痫大鼠不同脑区神经肽Y和亮氨酸–脑啡肽的表达及其意义
陈琅, 陈巧彬, 邹漳钰, 林立芳, 沈晓丽, 陈新, 季晓林   
  1. 350001 福州,福建省立医院儿科、福建医科大学省立临床医学院病理科
    福建省直机关医院神经科
    福建省心血管病重点实验室
  • 出版日期:2009-02-01

Expression of the Neuropeptide Y and Leu–Enkephalin in the Brain of Pentylenetrazol–Induced Epileptic Rat Models.

Lang CHEN, Qiao–bin CHEN, Zhang–yu ZOU, Li–fang LIN, Xiao–li SHEN, Xin CHEN, Xiao–lin JI   

  1. Department of Pediatrics, Fujian Provincial Hospital, Fuzhou 350001, China
  • Published:2009-02-01
  • Supported by:
    * Project No.Z0516068, supported by the Fujian Provincial Natural Science Funding
引用本文:

陈琅, 陈巧彬, 邹漳钰, 林立芳, 沈晓丽, 陈新, 季晓林. 戊四氮致癫痫大鼠不同脑区神经肽Y和亮氨酸–脑啡肽的表达及其意义[J]. 中华妇幼临床医学杂志(电子版), 2009, 05(01): 13-17.

Lang CHEN, Qiao–bin CHEN, Zhang–yu ZOU, Li–fang LIN, Xiao–li SHEN, Xin CHEN, Xiao–lin JI. Expression of the Neuropeptide Y and Leu–Enkephalin in the Brain of Pentylenetrazol–Induced Epileptic Rat Models.[J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2009, 05(01): 13-17.

目的

神经肽Y(neuropeptide Y, NPY)和亮氨酸–脑啡肽(leu–enkephalin, L–ENK)在癫痫发生、发展和转归中有着重要作用,本研究在戊四氮(pentylenetetrazol, PTZ)致癫痫大鼠模型中观察神经肽Y和亮氨酸–脑啡肽在不同脑区中的变化趋势,探讨神经肽Y、亮氨酸–脑啡肽与癫痫的关系。

方法

将清洁级近交系雄性SD大鼠50只分为实验组(A组,n=40)和对照组(B组,n=10)。A组大鼠腹腔注射戊四氮1次,剂量为50 mg/kg,B组腹腔注射同等容量生理盐水。A组大鼠腹腔注射戊四氮后,根据癫痫发作进行分级,0~1级(未发生惊厥)为7只(A1组,n=7),立即取脑处理;2级以上发作(大发作)为33只,随机于癫痫发作后立即取脑处理(A2组,n=10)及6 h( A3组,n=11),24 h( A4组,n=10),72 h(A5组,n=2)取脑处理。B组于注射生理盐水后5 min~10 min内取脑处理。采用放射免疫方法动态观察A,B两组大鼠脑组织的海马、中脑、纹状体和额叶中神经肽Y、亮氨酸–脑啡肽的改变;S–P免疫组化染色法染色,观察A,B两组大鼠海马CA1区神经肽Y、亮氨酸–脑啡肽的表达。

结果

A1组大鼠中脑、纹状体及额叶的神经肽Y含量较B组升高(P<0.05)。A,B两组海马CA1区神经肽Y含量比较,差异无显著意义(P<0.05)。癫痫大发作组(A2,A3和A4组)以上各部位神经肽Y含量均明显高于B组;动态观察结果显示,癫痫发作后,在中脑、海马、纹状体和额叶的神经肽Y含量明显增高,随后呈逐步下降趋势,癫痫发作后24 h,其含量与未发生惊厥的A1组神经肽Y含量比较,差异无显著意义(P>0.05) 。S–P免疫组化染色显示,癫痫大发作后,海马神经肽Y的表达明显增加,但随着时间的推移而降低。应用戊四氮后,癫痫大发作大鼠较未出现大发作的大鼠,其额叶、海马、中脑及纹状体中亮氨酸–脑啡肽含量均明显升高,两组间比较,除中脑外,其余部位差异有显著意义(P<0.05);但大发作大鼠海马、中脑及纹状体亮氨酸–脑啡肽含量与B组比较,高于B组,但差异无显著意义(P>0.05)。癫痫大发作后,各脑区亮氨酸–脑啡肽含量呈逐渐下降趋势。

结论

神经肽Y和亮氨酸–脑啡肽与癫痫的发生、发展有着密切关系。

Objective

To evaluate the significant changes of the neuropeptide Y(NPY) and leu-enkephalin(L-Enk) in the brain of pentylenetetrazol(PTZ)-induced rats in order to understand the relationship among the neuropeptide Y, leu-enkephalin and epilepsy.

Methods

Fifty male adult SD rats were randomly divided into the experimental group (group A, n=40, intraperitoneal pentylenetetrazol injection 50 mg/kg) and the control group (group B , n=10, intraperitoneal tales doses normal saline injection). According to the modification of seizure activity, the group A was divided into non-seizure group (group A1, n=7) which was decapitated immediately and the seizure group(group A2~A5, n=33). Rats of the seizure group were decapitated immediately (group A2, n=10), at the sixth hour(group A3, n=11), at the 24th hour(group A4, n=10) and at the 72th hour(group A5, n=2) after the seizure. Radioimmunoassay (RIA) and immunocytochemical techniques were used to evaluate the changes of the neuropeptide Y and leu-enkephalin in mesencephalon, hippocampus, striatum, and frontal cortex.

Results

The level of the neuropeptide Y increased significantly in mesencephalon(P<0.05), frontal cortex(P<0.05), and striatum (P<0.01) of the group A in contrast to those of the group B, and no difference was found in hippocampus. The level of the neuropeptide Y in brains of the seizure group(group A2~A5, n=33) was significant higher than that of the group B. The level of the neuropeptide Y returned to the value of the non-seizure group at the 24th hour after the seizure. Both groups had an elevated level of the leu-enkephalin in mesencephalon, striatum, hippocampus and frontal cortex. No difference was found between the non-convulsive group and the convulsive group in the four brain areas above, but the leu-enkephalin content of hypothalamus in the convulsive group was higher than that in the non-convulsive group. After the convulsion, the level of the leu-enkephalin decreased.

Conclusion

The increased level of the neuropeptide Y and leu-enkephalin may play an important role during the epilepsy.

1 Xie XK, Tang SM, Lv XL, et al. A study of opioid peptide in the CSF of epilepsy patients. Chin J Neuro Pshychia,1991,24:15–17.
2 Chen L, Zou ZY, Lin LF, et al. Expression of Leu–enkephalin in different cerebral regions of rats with pentylenetetrazol–induced epilepsy and its significance. Chin J Neuromed, 2005,4(8):783–789.[陈琅,邹漳钰,林立芳,等.实验性癫痫大鼠不同脑区L–ENK的表达及意义.中华神经医学杂志,2005,4(8):783–789.]
3 Tatemoto K, Carlquist M, Mutt V. Neuropeptide Y a novel brain peptide with structural similarities to peptide YY and pancreatic polypeptide. Nature, 1982,296:659–660.
4 Erickson JC, Clegg KE, Palmiter RD. Sensitivity to leptin and susceptibility to seizures of mice lacking neuropeptide Y. Nature, 1996,381(6581):415–418.
5 Reibel S, Larmet Y, Carnahan J, et al. Endogenous control of hippocampal epileptogenesis a molecular cascade involving brain–derived neurotrophic factor and neuropeptide Y. Epilepsia, 2000,41:s127–133.
6 Vezzani A, Ravizza T, Moneta D, et al. Brain–derived neurotrophic factor immunoreactivity in the limbic system of rats after acute seizures and during spontaneous convulsions: Temporal evolution of changes as compared to neuropeptide Y. Neuroscience, 1999,90(4): 1445–1461.
7 Woldbye DP, Larsen PJ, Mikkelsen JD, et al. Powerful inhibition of kainic acid seizures by neuropeptide Y via Y5–like receptors. Nat Med,1997,3(7):761–764.
8 DePrato PS, Holmes P, Martin R, et al. Experimentally induced attenuation of neuropeptide–Y gent expression in transgenic mice increase mortality rate following seizures. Neurosci Lett,2000,287:61–64.
9 Chen L, Ji XL, Zou ZY, et al. A study of the significance of various expression of Neuropeptide Y in brain of pentylenetrazol–induced rats. Chin J Neuroimmunol Neurol, 2005,12(2):118–121.[陈琅,季晓林,邹漳钰,等.戊四氮致痫大鼠不同脑区神经肽Y的表达及意义.中国神经免疫学和神经病学杂志,2005,12(2):118–121.]
10 Racine RJ. Modification of seizure activity by electrical stimulation.Ⅱ.Motor seizure. Electroencephalogr Clin Neurophysil,1972,32:281.
11 Klioueva IA, van Luijtelaar E,Chepurnova NE, et al. PTZ–induced seizures in rats: Effects of age and strain. Physiol Behav, 2001,72:421–426.
12 Corda MG, Orlandi M, Lecca D, et al. Pentylenetrazol–induced kindling in rats: Effect of GABA function inhibitors. Pharmacol Biochem Behav, 1991,40:329–333.
13 Rocha L, Cano A, Cruz C, et al. Opioid peptide systems following a subconvulsant dose of pentylenetetrazol in rats. Epilepsy Res,1999,37:141–150.
14 Vezzani A, Sperk G, Colmers WF. Neuropeptide Y: Emerging evidence for a functional role in seizure modulation. Trends Neurosci,1999,22:25–30.
15 Vezzani A, Michalkiewicz M, Moneta D, et al. Seizure susceptibility and epileptogenesis are decreased in transgenic rats overexpressing neuropeptide Y. Neurosci,2002,110:237–243.
16 Chen L, Ji XL. Enkephalin, neuropeptide Y, neuron–specific enolase and epilepsy. Foreign Med Infor, 2004,25(1):7–11.[陈琅,季晓林.脑啡肽、神经肽Y、神经元特异性烯醇酶与癫痫.国外医学情报,2004,25(1):7–11.]
17 Wu YF, Li SB.Neuropeptide Y expression in mouse hippocampus and its role in neuronal excitotoxicity. Acta Pharmacol Sinaca, 2005,26(1):63–68.
18 Vezzani A, Civenni G, Rizzi M, et al. Enhanced neuropeptide Y release in the hippocampus is associated with chronic seizure susceptibility in kainic acid treated rats. Brain Res, 1994,660(1):138–142.
19 Fu LY, Claudio AG, Anthony NP. Neuropeptide Y inhibits hypocretin/orexin neurons by multiple presynaptic and postsynaptic mechanisms: Tonic depression of the hypothalamic arousal system.J Neurosci, 2004, 24(40):8741–8751.
20 Baraban SC, Hollopeter G, Erickson JC, et al. Knock–out mice reveal a critical antiepileptic role for neuropeptide Y. J Neurosci, 1997,17(23):8927.
21 Woldbye DPD. Antiepilepsytic effects of NPY on pentylenetetrazole seizures. Regulat Peptid,1998,75–76:279–282.
22 Duan RS, Jin S, Chi ZF. Effects of lateral ventricle infusion with neuropeptide–Y on expression of brain–derived neurotrophic factor in hippocampus after pentylenetetrazol–induced epileptic seizure in rats. J Brain Nerv Dise, 2002,10(5):275–278.[段瑞生,金善,迟兆富.脑室内灌注神经肽Y对戊四氮致痫大鼠海马结构内脑源性神经营养因子表达的影响.脑与神经疾病杂志,2002,10(5):275–278.]
23 Patrylo PR, van Den Pol A, Spencer DD, et al. NPY inhibits glutamaterigic exicitation in the epileptic human dentate gyrus. J Neurophysiol, 1999,82(1):478–483.
24 Urca G, Frenk M. Intracerebral opiates blocks the epileptic effect of intracerebroventricular leucine–enkephalin. Brain Res, 1983,259:103–106.
25 Ishikura N, Tsunashima K, Watanabe K, et al. Temporal change of hippocampal enkephalin and dynorphin mRNA following trimethyltin intoxication in rats: Effect of anticonvulsant. Neurosci Lett,2001,306:157–160.
26 Rocha L, Maidment NT. Opioid peptide relaease in the rat hippocampus after kainic acid–induced status epilepticus. Hippocampus,2003,13:472–480.
27 Fetissov SO, Jacoby AS, Brumovsky PR, et al. Alter hippocampal expression of neuropeptides in seizure–prone GALR1 knockout mice. Epilepsia, 2003,44:1022–1033.
28 Harrison MB, Shumate MD, Lothman EN. Opioid peptide expression in models of chronic temporal lobe epilepsy. Neurosci, 1995,65:785–795.
29 Chen L, Zou ZY, Chen X, et al. The study of neuron damage of rat hippocampus after PTZ–induced seizure. J Brain Nerv Dise, 2005,13(3):200–203.[陈琅,邹漳钰,陈新,等.PTZ致痫大鼠海马区神经元损伤的研究.脑与神经疾病杂志,2005,13(3):200–203.]
30 Chen L, Zhao XZ, Zou ZY, et al. Changes of ultrastructure of rat hippocampus CA1 after PTZ–induced seizure. J Fujian Med Univ, 2004,38(4):448–450.[ 陈琅,赵晓贞,邹漳钰,等.戊四氮致痫大鼠海马CA1区超微结构改变.福建医科大学学报,2004,38(4):448–450.]
[1] 杨莹, 刘艳, 王央丹. 新生儿结节性硬化症相关性癫痫1例并文献复习[J]. 中华妇幼临床医学杂志(电子版), 2023, 19(04): 464-472.
[2] 邓瑶, 喻韬, 陈小璐, 罗蓉. 热性感染相关性癫痫综合征患儿的临床及随访分析[J]. 中华妇幼临床医学杂志(电子版), 2022, 18(06): 645-651.
[3] 罗序峰, 廖建湘, 罗智强, 段婧, 李永利, 徐建芳, 陈黎. Na+通道阻滞剂治疗SCN2A基因变异所致早发型癫痫性脑病并文献复习[J]. 中华妇幼临床医学杂志(电子版), 2022, 18(05): 585-590.
[4] 宁俊杰, 乔莉娜. 非典型良性家族性新生儿癫痫患儿的诊治及文献复习[J]. 中华妇幼临床医学杂志(电子版), 2022, 18(03): 307-314.
[5] 廖怡, 罗蓉. 儿童脑型X-连锁肾上腺脑白质营养不良分析并文献复习[J]. 中华妇幼临床医学杂志(电子版), 2022, 18(02): 205-212.
[6] 杨骐源, 张琼, 刘亚琪, 廖桉然, 邹静. 进行性半侧颜面萎缩伴癫痫1例[J]. 中华口腔医学研究杂志(电子版), 2023, 17(02): 89-94.
[7] 王利, 张磊, 费晓炜, 伊西才, 王彦刚. 迷走神经电刺激术中充分剥离迷走神经鞘膜对术后癫痫发作影响的研究[J]. 中华神经创伤外科电子杂志, 2023, 09(03): 160-164.
[8] 齐洪武, 刘岩松, 曾维俊, 张立钊, 郭洪均, 刘清石. 儿童创伤性脑损伤的研究进展[J]. 中华神经创伤外科电子杂志, 2022, 08(02): 121-124.
[9] 吴新高, 李伟, 马俊, 刘红, 黄平. 癫痫患者定量脑电图功率谱相关参数与认知功能损伤的相关性研究[J]. 中华脑科疾病与康复杂志(电子版), 2023, 13(05): 304-308.
[10] 郝梦薇, 陈珍, 魏涛, 张慧. 成年癫痫患者血清miR-132-3p表达及其与耐药的关系分析[J]. 中华脑科疾病与康复杂志(电子版), 2022, 12(06): 344-348.
[11] 丁浩然, 关宇光, 王雄飞, 赵萌, 王静, 王梦阳, 滕鹏飞, 栾国明. 立体定向脑电图引导下多电极立体适形射频热凝毁损治疗药物难治性岛叶癫痫的疗效分析[J]. 中华脑科疾病与康复杂志(电子版), 2022, 12(04): 227-233.
[12] 贺晶, 史洁, 林久銮, 孙朝晖, 王海祥, 张冰清, 刘一鸥, 宋宪成, 丰倩, 柏建军, 周文静. 结节性硬化伴难治性癫痫儿童的手术治疗分析[J]. 中华临床医师杂志(电子版), 2021, 15(08): 597-600.
[13] 王媛媛, 冯兆才, 路岩莉. 头部经络与癫痫的关系研究进展[J]. 中华针灸电子杂志, 2022, 11(02): 55-57.
[14] 许洁, 李秋波, 孔庆霞, 郑明雪. 局灶性皮层发育不良发病机制研究进展[J]. 中华诊断学电子杂志, 2022, 10(01): 31-35.
[15] 李承玉, 徐连萍, 王圣松, 王群. 不同抗癫痫发作药物单药治疗在卒中后癫痫中的保留率和有效性分析[J]. 中华脑血管病杂志(电子版), 2023, 17(04): 325-330.
阅读次数
全文


摘要