中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2009年
23期
4559-4562
,共4页
张文平%张亘瑷%仇玉兰%张策
張文平%張亙璦%仇玉蘭%張策
장문평%장긍애%구옥란%장책
神经发生%长时程增强%Ro25-6981%神经元%齿状回
神經髮生%長時程增彊%Ro25-6981%神經元%齒狀迴
신경발생%장시정증강%Ro25-6981%신경원%치상회
In the rat hippocampal dentate gyrus%NR2B receptor antagonist Ro25-6981 blocked ACSF-LTP and partly inhibited BIC-LTP%indicating that NR2B receptor antagonist can inhibit newborn neuron-induced LTP
背景:新生神经元可诱导产生长时程增强,NMDA受体亚基NR2B的激活在成熟神经元诱导的长时程增强中有重要作用,但其对由新生神经元诱导的长时程增强的影响尚未见报道.目的:观察NR2B受体拮抗剂Ro25-6981在大鼠齿状回新生神经元诱导的长时程增强中的作用.设计、时间及地点:脑片电生理实验,于2007-02/06在山西医科大学神经生物实验室完成.材料:3月龄雄性Wistar大鼠26只,由山西医科大学实验动物中心提供.方法:大鼠麻醉后断头取脑,分离海马,制备400μmol/L脑片.采用细胞外微电极记录技术,于海马齿状回分子层内侧1/3处采用双极钨电极进行低频刺激,获得稳定的刺激曲线后,在高频强直刺激下诱导长时程增强.长时程增强的诱导程序为每串刺激频率100 Hz,持续500 ms,间隔20 s,共4串刺激,记录到兴奋性突触后电位＞1 mV以上的脑片用于下述两项实验.①NR2B拮抗剂R025-6981阻断人工脑脊液诱导的长时程增强:脑片分为2组,人工脑脊液组持续通以含有95%O2和5%CO2的人工脑脊液,人工脑脊液+Ro25-6981组在强直刺激前应用3 μmol/L NR2B拮抗剂Ro25-6981作用10 min,后续步骤同上组.②NR2B拮抗剂Fo25-6981抑制荷包牡丹碱诱导的长时程增强:脑片分为2组,荷包牡丹碱组在强直刺激前给予10 μ mol/L荷包牡丹碱灌流10 min,荷包牡丹碱+Ro25-6981组在强直刺激前同时给予3 μ mol/L Ro25-6981和10 μmol/L荷包牡丹碱灌流10 min.主要观察指标:长时程增强记录结果.结果:①强直刺激后50～60 min,人工脑脊液组长时程增强(107.85±1.34)%,人工脑脊液+Ro25-6981组长时程增强(100.75±2.75)%,两组比较差异有显著性意义(P＜0.05).②强直刺激后50～60 min,荷包牡丹碱组长时程增强(164.67±2.40)%,荷包牡丹碱+Ro25-6981组长时程增强(147.56±6.63)%,两组比较差异有显著性意义(P＜0.05).结论:在大鼠海马齿状回区域,NR2B受体拮抗剂Ro25-6981阻断了人工脑脊液诱导的长时程增强,并部分抑制了荷包牡丹碱诱导的长时程增强,提示NR2B受体拮抗荆可以抑制新生神经元诱导的长时程增强.
揹景:新生神經元可誘導產生長時程增彊,NMDA受體亞基NR2B的激活在成熟神經元誘導的長時程增彊中有重要作用,但其對由新生神經元誘導的長時程增彊的影響尚未見報道.目的:觀察NR2B受體拮抗劑Ro25-6981在大鼠齒狀迴新生神經元誘導的長時程增彊中的作用.設計、時間及地點:腦片電生理實驗,于2007-02/06在山西醫科大學神經生物實驗室完成.材料:3月齡雄性Wistar大鼠26隻,由山西醫科大學實驗動物中心提供.方法:大鼠痳醉後斷頭取腦,分離海馬,製備400μmol/L腦片.採用細胞外微電極記錄技術,于海馬齒狀迴分子層內側1/3處採用雙極鎢電極進行低頻刺激,穫得穩定的刺激麯線後,在高頻彊直刺激下誘導長時程增彊.長時程增彊的誘導程序為每串刺激頻率100 Hz,持續500 ms,間隔20 s,共4串刺激,記錄到興奮性突觸後電位＞1 mV以上的腦片用于下述兩項實驗.①NR2B拮抗劑R025-6981阻斷人工腦脊液誘導的長時程增彊:腦片分為2組,人工腦脊液組持續通以含有95%O2和5%CO2的人工腦脊液,人工腦脊液+Ro25-6981組在彊直刺激前應用3 μmol/L NR2B拮抗劑Ro25-6981作用10 min,後續步驟同上組.②NR2B拮抗劑Fo25-6981抑製荷包牡丹堿誘導的長時程增彊:腦片分為2組,荷包牡丹堿組在彊直刺激前給予10 μ mol/L荷包牡丹堿灌流10 min,荷包牡丹堿+Ro25-6981組在彊直刺激前同時給予3 μ mol/L Ro25-6981和10 μmol/L荷包牡丹堿灌流10 min.主要觀察指標:長時程增彊記錄結果.結果:①彊直刺激後50～60 min,人工腦脊液組長時程增彊(107.85±1.34)%,人工腦脊液+Ro25-6981組長時程增彊(100.75±2.75)%,兩組比較差異有顯著性意義(P＜0.05).②彊直刺激後50～60 min,荷包牡丹堿組長時程增彊(164.67±2.40)%,荷包牡丹堿+Ro25-6981組長時程增彊(147.56±6.63)%,兩組比較差異有顯著性意義(P＜0.05).結論:在大鼠海馬齒狀迴區域,NR2B受體拮抗劑Ro25-6981阻斷瞭人工腦脊液誘導的長時程增彊,併部分抑製瞭荷包牡丹堿誘導的長時程增彊,提示NR2B受體拮抗荊可以抑製新生神經元誘導的長時程增彊.
배경:신생신경원가유도산생장시정증강,NMDA수체아기NR2B적격활재성숙신경원유도적장시정증강중유중요작용,단기대유신생신경원유도적장시정증강적영향상미견보도.목적:관찰NR2B수체길항제Ro25-6981재대서치상회신생신경원유도적장시정증강중적작용.설계、시간급지점:뇌편전생리실험,우2007-02/06재산서의과대학신경생물실험실완성.재료:3월령웅성Wistar대서26지,유산서의과대학실험동물중심제공.방법:대서마취후단두취뇌,분리해마,제비400μmol/L뇌편.채용세포외미전겁기록기술,우해마치상회분자층내측1/3처채용쌍겁오전겁진행저빈자격,획득은정적자격곡선후,재고빈강직자격하유도장시정증강.장시정증강적유도정서위매천자격빈솔100 Hz,지속500 ms,간격20 s,공4천자격,기록도흥강성돌촉후전위＞1 mV이상적뇌편용우하술량항실험.①NR2B길항제R025-6981조단인공뇌척액유도적장시정증강:뇌편분위2조,인공뇌척액조지속통이함유95%O2화5%CO2적인공뇌척액,인공뇌척액+Ro25-6981조재강직자격전응용3 μmol/L NR2B길항제Ro25-6981작용10 min,후속보취동상조.②NR2B길항제Fo25-6981억제하포모단감유도적장시정증강:뇌편분위2조,하포모단감조재강직자격전급여10 μ mol/L하포모단감관류10 min,하포모단감+Ro25-6981조재강직자격전동시급여3 μ mol/L Ro25-6981화10 μmol/L하포모단감관류10 min.주요관찰지표:장시정증강기록결과.결과:①강직자격후50～60 min,인공뇌척액조장시정증강(107.85±1.34)%,인공뇌척액+Ro25-6981조장시정증강(100.75±2.75)%,량조비교차이유현저성의의(P＜0.05).②강직자격후50～60 min,하포모단감조장시정증강(164.67±2.40)%,하포모단감+Ro25-6981조장시정증강(147.56±6.63)%,량조비교차이유현저성의의(P＜0.05).결론:재대서해마치상회구역,NR2B수체길항제Ro25-6981조단료인공뇌척액유도적장시정증강,병부분억제료하포모단감유도적장시정증강,제시NR2B수체길항형가이억제신생신경원유도적장시정증강.
BACKGROUND: Newborn neurons have bean shown to induce long-term potentiation (LTP). Activation of N-methyl-D-aspartic acid (NMDA) receptor subunit NR2B plays an important role in mature neurons-induced LTP. But there have been no reports addressing on the effects of NR2B activation on newborn neuron-induced LTP.OBJECTIVE: To investigate the effects of NR2B receptor antagonist Ro25-6981 on LTP induced by newborn neurons in adult rat dentate gyrus.DESIGN, TIME AND SETTING: An electrophysiological recording trial was performed at the Department of Neuroblology,Shanxi Medical University from February to June 2007.MATERIALS: Twenty-six male Wistar rats, aged 3 months, were provided by Laboratory Animal Center, Shanxi Medical University.METHODS: Following sacrifice for brain harvesting under anesthesia, the hippocempus was taken to preparation of 400 μ mol/L brain slices. Using extracellular field potential recordings, low-frequency stimulation was performed in the medial molecular layer of dentate gyrus with insulated bipolar tungsten electrodes. After having stable recordings, LTP was induced under high-frequency tetanic stimulation. LTP was induced with a protocol developed previously (4 trains, 500 ms each, 100 Hz within the train, repeated every 20 s). Only those slices which produced the field excitatory postsynaptic potential of 1 mV or cerebrospinal fluid (ACSF)-induced LTP (ACSF-LTP): brain slices were divided into 2 groups: ACSF group, in which, slices were continuously perfused using ACSF bubbled with 95% O2 and 5% CO2; ACSF+ Ro25-6981 group: a 10-minute treatment with 3μ mol/L Ro25-6981 was performed prior to tetanic stimulation, and the remaining procedures were the same as ACSF divided into 2 groups: BIC group: a 10-minute treatment with 10 μmol/L BIC was performed prior to titanic stimulation; BIC+Ro25-6981 group: 3μ mol/L Ro25-6981 and 10μ mol/L BIC were simultaneously perfused 10 minutes prior to tetanic stimulation.MAIN OUTCOME MEASURES: LTP recording results.minutes of titanic stimulation, LTP was (164.67±2.40)% and (147.56±6.63)% in the BIC and BIC+ Ro25-6981 groups,respectively, and a significant difference existed between the two groups (P ＜ 0.05).
