本文選題：蒼白球 + orexin-A　； 參考：《青島大學(xué)》2017年碩士論文

【摘要】：Orexins是下丘腦神經(jīng)肽家族中的一員,在中樞神經(jīng)系統(tǒng)的運動調(diào)節(jié)功能中起重要作用。已有文獻證明,帕金森病患者和動物模型腦內(nèi)orexin水平明顯降低。蒼白球是基底神經(jīng)節(jié)間接通路的重要核團,在機體運動協(xié)調(diào)等方面發(fā)揮重要作用,其功能異常與許多運動功能障礙疾病相關(guān)。形態(tài)學(xué)研究表明,蒼白球接受orexin纖維投射并表達orexin-1受體(OX1R)。目的:探究在正常及MPTP(1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)帕金森病模型小鼠,蒼白球orexin-A的電生理效應(yīng)及其受體機制。方法:本實驗采用多管微電極在體細胞外電生理記錄、MPTP帕金森病小鼠模型制備以及免疫組織化學(xué)染色等實驗方法。結(jié)果:1.在體細胞外電生理實驗中記錄的59個正常小鼠蒼白球神經(jīng)元,平均放電頻率為8.43±0.55 Hz。蒼白球神經(jīng)元呈現(xiàn)三種放電模式,其中29個(49.15%)呈規(guī)則放電,22個(37.29%)呈不規(guī)則放電,8個(13.56%)呈簇狀放電。2.在正常小鼠記錄到38個蒼白球神經(jīng)元,有21個神經(jīng)元在微量壓力注射orexin-A(0.01m M)后自發(fā)放電頻率由7.77±0.94 Hz升高到11.10±1.23 Hz,平均升高51.26±6.36%(t=-7.20,P=0.000)。經(jīng)過對放電模式CV值和FF值分析,orexin-A使CV值增加(加藥前:0.50±0.09;加藥后:0.63±0.09),差異有統(tǒng)計學(xué)意義(t=-2.48,P=0.02),對FF值無明顯影響(加藥前:0.12±0.04;加藥后:0.11±0.04,t=0.47,P=0.65)。3.正常小鼠蒼白球微量注射OX1R阻斷劑SB-334867(0.01m M),在記錄到的33個神經(jīng)元中,有23個自發(fā)放電頻率由8.71±0.95 Hz降低至3.50±0.63 Hz,平均降低61.77±5.42%,加藥前與加藥后比較,差異有顯著統(tǒng)計學(xué)意義(t=7.69,P=0.000)。SB-334867使CV值增加(加藥前:0.89±0.12;加藥后:1.17±0.12;t=-3.32,P=0.003),并使FF值增加(加藥前:0.21±0.07;加藥后:0.70±0.16;t=-3.52,P=0.002),差異均有統(tǒng)計學(xué)意義。該結(jié)果提示蒼白球orexin-A遞質(zhì)系統(tǒng)能夠改變蒼白球神經(jīng)元放電模式。4.正常小鼠記錄到7個蒼白球神經(jīng)元,聯(lián)合微量注射0.01m M SB-334867和0.01m M orexin-A,使蒼白球神經(jīng)元放電頻率自4.66±1.28Hz升高至5.12±1.34 Hz,平均升高13.16±3.19%。待完全恢復(fù)后,在同一神經(jīng)元中再單獨給予0.01m M orexin-A,蒼白球神經(jīng)元放電頻率自5.15±1.16Hz升高至7.40±1.68 Hz,平均升高46.43±7.22%,與之前聯(lián)合給予SB-334867和orexin-A組比較差異有統(tǒng)計學(xué)意義(z=-2.49,P=0.011)。5.在MPTP帕金森病模型小鼠記錄到的33個蒼白球神經(jīng)元,自發(fā)放電頻率平均為7.64±0.96 Hz,與正常小鼠相比無明顯差異(t=0.42,P=0.68)。放電模式分析顯示12個(36.36%)神經(jīng)元呈規(guī)則放電,16個(48.49%)神經(jīng)元呈不規(guī)則放電,5個(15.15%)神經(jīng)元呈簇狀放電。MPTP帕金森病模型小鼠蒼白球神經(jīng)元的規(guī)則放電百分比較正常小鼠有所減少,而不規(guī)則放電以及簇狀放電的百分比有所增加,但χ2檢驗結(jié)果顯示差異無顯著性(χ2=1.42,d.f.=2,P0.05)。6.在MPTP帕金森病模型小鼠,蒼白球微量壓力注射0.01m M orexin-A可以使記錄到的23個神經(jīng)元中的14個神經(jīng)元產(chǎn)生興奮效應(yīng),使其自發(fā)放電頻率由6.14±1.17 Hz升高到10.40±1.82 Hz,平均升高83.78±16.82%(t=-5.20,P=0.000)。對放電模式CV值(加藥前:0.49±0.09;加藥后:0.60±0.11;t=-1.40,P=0.19)和FF值(加藥前:0.16±0.08;加藥后:0.13±0.07;t=0.22,P=0.83)分析,差異無統(tǒng)計學(xué)意義。有2個蒼白球神經(jīng)元在微量壓力注射0.01m M orexin-A后自發(fā)放電頻率由5.95±2.58 Hz降低到3.12±1.02 Hz,平均降低44.58±6.89%(t=1.81,P=0.32)。7.帕金森病模型小鼠蒼白球微量注射OX1R阻斷劑SB-334867(0.01m M),在記錄到的10個神經(jīng)元中,有7個自發(fā)放電頻率由6.71±1.47 Hz降低至2.49±1.09 Hz,平均降低68.84±8.40%,加藥前與加藥后比較,差異有顯著統(tǒng)計學(xué)意義(t=5.91,P=0.001)。SB-334867使CV值增加(加藥前:0.99±0.12;加藥后:1.33±0.12;t=-6.09,P=0.001),并使FF值增加(加藥前:0.18±0.04;加藥后:0.61±0.11;t=-5.60,P=0.001),差異均有統(tǒng)計學(xué)意義。8.Orexin-A對MPTP帕金森病模型小鼠蒼白球神經(jīng)元的興奮效應(yīng)較正常小鼠明顯增強,差異有統(tǒng)計學(xué)意義(z=-2.02,P=0.04);而SB-334867對正常小鼠和MPTP帕金森病模型小鼠蒼白球神經(jīng)元的抑制效應(yīng)比較,差異無統(tǒng)計學(xué)意義(z=-0.19,P=0.85)。9.免疫組織化學(xué)染色顯示正常和MPTP帕金森病模型小鼠蒼白球均可表達OX1R,且表達量相當(dāng)(P0.05)。兩種小鼠下丘腦外側(cè)區(qū)的orexin-A的表達量也無顯著差異(P0.05)。結(jié)論:Orexin-A通過激活OX1R提高正常和MPTP帕金森病模型小鼠蒼白球神經(jīng)元興奮性,且orexin-A對帕金森病模型小鼠蒼白球神經(jīng)元的興奮效應(yīng)較正常小鼠增強。SB-334867能夠抑制正常和帕金森病模型小鼠蒼白球神經(jīng)元放電頻率,提示內(nèi)源性orexin主要通過OX1R發(fā)揮作用。本實驗為蒼白球orexin遞質(zhì)系統(tǒng)參與帕金森病發(fā)病提供了一定的理論和實驗依據(jù)。
[Abstract]:Objective : To investigate the electrophysiological effects of orexin - A and orexin - 1 receptor ( OX1R ) in normal and MPTP ( 1 - Methyl - 4 - phenyl - 1,2,3,6 - tetrahydropyridine , MPTP ) Parkinson ' s disease . The results showed that : 1 . In normal and MPTP ( 1 - Methyl - 4 - phenyl - 1,2,3,6 - tetrahydropyridine , MPTP ) Parkinson ' s disease model mice , the frequency of spontaneous discharge was increased from 7.77 鹵 0.94 Hz to 11.10 鹵 1.23 Hz , with an average increase of 51.26 鹵 6.36 % ( t = - 7.20 , P = 0.000 ) . In the 33 neurons recorded , 23 spontaneous discharge frequencies were reduced from 8.71 鹵 0.95 Hz to 3.50 鹵 0.63 Hz , the mean decrease was 61.77 鹵 5.42 % , and the difference was statistically significant ( t = 7.69 , P = 0.000 ) . The results suggested that the neurons of the pallidosphere orexin - A were able to change the discharge pattern of the pallidus . The results suggested that the neurons of the pallidosphere orexin - A were able to change the discharge pattern of the pallidus . The results suggested that the neurons of the pallidosphere orexin - A could change the discharge pattern of the pallidus . The results suggested that the neurons of the pallidosphere orexin - A could change the discharge frequency of the neurons from 4.66 鹵 1.28Hz to 5.12 鹵 1.68Hz , the average increase was 13.16 鹵 3.19 % . The results showed that 12 ( 36.36 % ) neurons showed regular discharge , 16 ( 48.49 % ) neurons were irregular discharges , 5 ( 15.15 % ) neurons were in cluster - like discharge . The percentage of irregular discharges and cluster - like discharges increased significantly in MPTP Parkinson ' s disease model mice , and the results showed no significant difference ( 蠂 ~ 2 = 1.42 , d.f . = 2 , P0.05 ) . The frequency of spontaneous discharge was reduced from 5.95 鹵 2.58 Hz to 3.12 鹵 1.02 Hz , with an average decrease of 44.58 鹵 6.89 % ( t = 1.81 , P = 0.32 ) . The results showed that orexin - A significantly increased the expression of orexin - A in normal and MPTP Parkinson ' s disease model mice .

【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R742.5;R-332

【參考文獻】

相關(guān)期刊論文 前1條

1 崔龍彪;李博為;金曉航;趙琳;史娟;;6-羥多巴誘致帕金森病大鼠模型中orexin系統(tǒng)的進行性變化(英文)[J];Neuroscience Bulletin;2010年05期

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本文編號：1797821