[1]张博爱)△,孙桂芳),贾延劼),等.OrexinA对体外培养的大鼠海马神经细胞凋亡的影响*[J].郑州大学学报(医学版),2009,(05):968-970.
 ZHANG Boai),SUN Guifang),JIA Yanjie),et al.Effect of OrexinA on apoptosis of rat hippocampal neurons cultured in vitro[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2009,(05):968-970.
点击复制

OrexinA对体外培养的大鼠海马神经细胞凋亡的影响*()
分享到:

《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2009年05期
页码:
968-970
栏目:
论著
出版日期:
2009-09-01

文章信息/Info

Title:
Effect of OrexinA on apoptosis of rat hippocampal neurons cultured in vitro
作者:
张博爱1)孙桂芳1)贾延劼1)董为伟2)谢鹏2)
1)郑州大学第一附属医院神经内科郑州4500522)重庆医科大学神经病学研究所重庆410008
Author(s):
ZHANG Bo’ai1)SUN Guifang1)JIA Yanjie1) DONG Weiwei2) XIE Peng2)
1)Department of Neurology, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052 2)Neurological Research Institute,Chongqing Medical College,Chongqing 410008
关键词:
神经细胞海马OrexinAOX1受体大鼠
Keywords:
neuronhippocampusOrexinAOX1 receptorrat
分类号:
Q255
摘要:
研究OrexinA对体外培养的海马神经细胞凋亡的影响及其可能机制。方法:取体外培养的大鼠海马神经细胞分为7组,分别给予0,1.0,10.0,100.0nmol/L及1.0、10.0、100.0μmol/L的OrexinA培养。另取大鼠海马神经细胞,预先加入OrexinA受体(OX1R)阻断剂SB334867,然后同法分组及处理。观察海马神经细胞的形态学变化;采用WesternBlot、逆转录PCR(RTPCR)法检测OX1R蛋白和mRNA的表达;原位末端标记染色评价细胞凋亡率;Fura2/AM荧光比值成像技术测定游离钙离子浓度[Ca2+]i。结果:随着OrexinA浓度的增大,海马神经细胞凋亡增加(F=137.49,P<0.01),同时呈剂量依赖性地触发[Ca2+]i的升高(F=961.44,P<0.01),但是OX1R蛋白及mRNA的表达无明显变化。SB334867可以部分阻断这种现象。结论:大剂量OrexinA对体外培养的海马神经细胞有促凋亡作用,其机制可能与增加了细胞内Ca2+浓度蓄积有关
Abstract:
To investigate the effect of OrexinA on apoptosis of rat hippocampal neurons cultured in vitro.Methods:The rat hippocampal neurons were allocated into 7 groups and cultured with 0,1.0,10.0,100.0 nmol/L and 1.0,10.0,100.0 μmol/L OrexinA.Then another rat hippocampal neurons were allocated into 7 groups and treated with SB334867 to block OX1R in advance,and then cultured with various doses of OrexinA as above. The morphological changes of neurons were observed.The expressions of OX1R protein and mRNA were assessed by Western blot and RTPCR. The percentage of apoptotic cells was detected by TUNEL assay.The concentration of [Ca2+]i was determined by spectrofluorometer and fluorescent Ca2+ probe Fura2/Am.Results: With the increase of the OrexinA concentrations,the cell apoptosis and [Ca2+]i both increased (F=137.49 and 961.44,P<0.01). However,OrexinA did not alter the expression of OX1R protein and mRNA. Pretreatment with SB334867, an OX1R antagonist,significantly antagonized this OrexinAinduced neurotoxicity on hippocampal neurons.Conclusion: The results suggest that treatment with the highdose OrexinA may produce a proapoptotic effect on normal hippocampal neurons, which is possibly involved in the increase of intracellular calcium.

参考文献/References:

[1]Ferguson AV, Samson WK. The orexin/hypocretin system: a critical regulator of neuroendocrine and autonomic function[J]. Front Neuroendocrinol,2003,24(3):141
[2]Irving EA, Harrison DC, Babbs AJ, et al. Increased cortical expression of the orexin1 receptor following permanent middle cerebral artery occlusion in the rat[J]. Neurosci Lett, 2002,324(1):53
[3]Zhao YY, Guo L, Du J, et al. Effects of acute hypoglycemia on the orexin system in rat[J]. Chin Med Sci J, 2005, 20(1):55
[4]Brewer GJ.Isolation and culture of adult rat hippocampal neurons [J]. J Neurosci Methods, 1997,71(2):145
[5]Smart D, Jerman J. The physiology and pharmacology of the orexins[J].Pharmacol Ther, 2002,94(1/2): 51
[6]Neves SR, Ram PT, Iyengar R. G protein pathways[J]. Science, 2002,296(5 573):1 636
[7]Nakamura T, Uramura K, Nambu T, et al. Orexininduced hyperlocomotion and stereotypy are mediated by the dopaminergic system [J]. Brain Res, 2000,873(1):181
[8]Smart D, Jerman JC, Brough SJ, et al. Characterization of recombinant human orexin receptor pharmacology in a Chinese hamster ovary cellline using FLIPR[J]. Br J Pharmacol,1999,128(1):1
[9]Rami A, Agarwal R, Botez G, et al. muCalpain activation,DNA fragmentation, and synergistic effects of caspase and calpain inhibitors in protecting hippocampal neurons from ischemic damage[J]. Brain Res, 2000, 866 (1/2): 299
[10]RouetBenzineb P, RouyerFessard C, Jarry A, et al. Orexins acting at native OX(1) receptor in colon cancer and neuroblastoma cells or at recombinant OX(1) receptor suppress cell growth by inducing apoptosis[J]. J Biol Chem, 2004, 279(44): 45 875
[11]Trivedi P, Yu H, MacNeil DJ, et al. Distribution of orexin receptor mRNA in the rat brain [J].FEBS Lett,1998,438(1/2):71
[12]Langmead CJ, Jerman JC, Brough SJ, et al. Characterisation of the binding of [3H]SB674042, a novel nonpeptide antagonist, to the human orexin1 receptor[J]. Br J Pharmacol, 2004,141(2):340

相似文献/References:

[1]黄静)△,方琴),王季石).环境激发吗啡条件性位置偏爱重现大鼠海马内脑源性神经营养因子及cfos的表达*[J].郑州大学学报(医学版),2010,(02):198.
 HUANG Jing),FANG Qin),WANG Jishi).Expression of braindrived neurophic factor and cfos in hippocampus of rats when morphine CPP was reinstated by environment[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2010,(05):198.
[2]蒋洪燕,李丽,郑艳涛,等.母体铅暴露对仔鼠海马组织中糖原合成酶激酶3β mRNA表达的影响[J].郑州大学学报(医学版),2010,(05):815.
 JIANG Hongyan,LI Li),ZHENG Yantao),et al.Effects of maternal lead exposure on the expression of glycogen synthase kinase3β mRNA in hippocampus of mouse pups[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2010,(05):815.
[3]钟志超,范洪领,尹金宝,等.氯通道电流在离体大鼠海马缺血缺氧性神经元凋亡中的变化及SITS的拮抗作用*[J].郑州大学学报(医学版),2011,(03):367.
 ZHONG Zhichao,FAN Hongling,YIN Jinbao,et al. Changes of chloride channel currents during cultured rat hippocampal neuronal apoptosis and antieffects of SITS[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2011,(05):367.
[4]陈灿映,冯静,刘芳丽,等.孕哺期铅暴露对仔鼠学习记忆能力及海马组织中PJNK表达的影响[J].郑州大学学报(医学版),2012,(01):78.
 CHEN Canying,FENG Jing,LIU Fangli,et al.Effects of maternal lead exposure on learning and memory and the expression of phosphorylated cJun Nterminal kinase in mice offspring[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2012,(05):78.

备注/Memo

备注/Memo:
*国家自然科学基金资助项目30770758 △女,1953年生,大学本科,教授,研究方向:神经病学基础与临床
更新日期/Last Update: 2010-05-14