發(fā)布時間：2018-04-08 21:01

  本文選題：Orexin　切入點：黑質(zhì)致密帶　出處：《青島大學(xué)》2014年碩士論文

【摘要】：黑質(zhì)是基底神經(jīng)節(jié)的重要核團之一,在機體運動功能調(diào)節(jié)中發(fā)揮重要作用。Orexin是下丘腦神經(jīng)肽家族成員,可直接參與調(diào)節(jié)中樞運動功能。形態(tài)學(xué)研究證實,黑質(zhì)接受大量來自下丘腦的orexin能纖維支配,并表達高水平的orexin1和orexin2(OX1和OX2)受體。帕金森病晚期患者下丘腦orexin能神經(jīng)元缺失,腦脊液orexin水平降低,且orexin的水平與帕金森病嚴(yán)重程度呈負相關(guān)。目的：本實驗探究orexin-A和OX1受體阻斷劑SB-334867對正常和6-羥基多巴胺(6-hydroxydopamine,6-OHDA)帕金森病模型大鼠黑質(zhì)神經(jīng)元電活動的調(diào)節(jié),以及正常和帕金森病大鼠黑質(zhì)OX1受體的表達。方法：本實驗采用多管微電極在體細胞外電生理記錄、6-OHDA帕金森病模型大鼠的制備、核團埋管術(shù)以及免疫組織化學(xué)染色等實驗方法。結(jié)果：1.正常大鼠黑質(zhì)致密帶微壓力注射0.01mM的orexin-A,在記錄到的58個多巴胺能神經(jīng)元中,有36個(62.1%)放電頻率明顯升高,自發(fā)放電頻率由2.3±0.3Hz升高至4.2±0.5Hz,平均升高101.0±18.0%(P0.001)。在正常大鼠黑質(zhì)網(wǎng)狀帶共記錄到54個GABA能神經(jīng)元,微壓力注射orexin-A可使其中24個(44.4%)神經(jīng)元放電頻率明顯升高,自發(fā)放電頻率由24.2±2.0Hz升高至31.4±2.7Hz,平均升高31.3±5.3%(P0.001)。2.在正常大鼠黑質(zhì)致密帶微壓力注射0.1mM的SB-334867,在記錄到的30個多巴胺能神經(jīng)元中,有19個(63.3%)放電頻率明顯降低,自發(fā)放電頻率由3.1±0.5Hz降低至1.0±0.3Hz,平均降低66.3±6.7%(P0.001);在黑質(zhì)網(wǎng)狀帶記錄到的21個GABA能神經(jīng)元中,SB-334767可使其中10個(47.6%)神經(jīng)元放電頻率明顯降低,自發(fā)放電頻率由21.9±3.0Hz降低至16.2±3.4Hz,平均降低28.44±7.9%(P0.01)。3在6-OHDA帕金森病模型大鼠損毀側(cè),微壓力注射0.01mM的orexin-A,可使黑質(zhì)致密帶記錄到的21個多巴胺能神經(jīng)元中的12個(57.1%)放電頻率明顯升高,由4.1±0.6Hz升高到6.5±1.0Hz,平均升高58.2±11.8%(P0.01)。在記錄到的12個黑質(zhì)網(wǎng)狀帶GABA能神經(jīng)元中,orexin-A可使其中7個(58.3%)神經(jīng)元放電頻率明顯升高,由19.6±2.7Hz升高到23.5±3.8Hz,平均升高18.8±3.0%(P0.05)。在正常和6-OHDA帕金森病模型大鼠,orexin-A對黑質(zhì)致密帶多巴胺能神經(jīng)元的興奮效應(yīng)較黑質(zhì)網(wǎng)狀帶強(正常大鼠P0.01；帕金森病模型大鼠P0.05)。4.免疫組織化學(xué)染色顯示,正常及帕金森病模型大鼠黑質(zhì)均表達OX1受體。 結(jié)論：Orexin-A可提高黑質(zhì)致密帶多巴胺能神經(jīng)元和網(wǎng)狀帶GABA能神經(jīng)元興奮性。內(nèi)源性O(shè)X1受體參與調(diào)節(jié)黑質(zhì)致密帶多巴胺能神經(jīng)元和網(wǎng)狀帶GABA能神經(jīng)元興奮性。在帕金森病模型大鼠,orexin-A也可使黑質(zhì)致密帶多巴胺能神經(jīng)元和網(wǎng)狀帶GABA能神經(jīng)元興奮性升高。正常及帕金森病模型大鼠黑質(zhì)均表達OX1受體。 本實驗結(jié)果為深入探討黑質(zhì)orexin-A在帕金森病發(fā)病中的作用提供了一定的理論和實驗依據(jù)。
[Abstract]:Substantia nigra is one of the most important nuclei in basal ganglia and plays an important role in the regulation of motor function.Morphological studies demonstrated that the substantia nigra was innervated by a large number of orexin energy fibers from the hypothalamus and expressed high levels of orexin1 and orexin2(OX1 and OX2) receptors.Orexin neurons were absent in hypothalamus and orexin level in cerebrospinal fluid was decreased in patients with advanced Parkinson's disease. The level of orexin was negatively correlated with the severity of Parkinson's disease.Aim: to investigate the effects of orexin-A and OX1 receptor blocker SB-334867 on the electrical activity of substantia nigra neurons in normal and 6-hydroxydopamine 6-hydroxydopamine 6-OHDA-Parkinson disease rats and the expression of OX1 receptors in the substantia nigra of normal and Parkinson's disease rats.Methods: the preparation of 6-OHDA Parkinson's disease model rats with multi-tube microelectrode in vitro electrophysiological recording, the implantation of nuclear tube and immunohistochemical staining were used in this experiment.The result is 1: 1.Of the 58 dopaminergic neurons recorded, 36 of the 58 dopaminergic neurons in the normal substantia nigra were injected with orexin-A under micropressure. The spontaneous discharge frequency increased from 2.3 鹵0.3Hz to 4.2 鹵0.5Hz.A total of 54 GABA neurons were recorded in the substantia nigra reticularis in normal rats. The frequency of spontaneous discharge increased from 24.2 鹵2.0Hz to 31.4 鹵2.7Hz. the frequency of spontaneous discharge increased from 24.2 鹵2.0Hz to 31.4 鹵2.7Hz.In normal rat substantia nigra, the discharge frequency of SB-334867 was significantly decreased in 19 of 30 dopaminergic neurons recorded by micropressure injection of 0.1mM.The frequency of spontaneous discharge decreased from 3.1 鹵0.5Hz to 1.0 鹵0.3 Hz, and decreased by 66.3 鹵6.7 Hz on average, and SB-334767 significantly decreased the firing frequency of 10 neurons among 21 GABA neurons recorded in the substantia nigra reticularis.The frequency of spontaneous discharge decreased from 21.9 鹵3.0Hz to 16.2 鹵3.4 Hz, and decreased by 28.44 鹵7.9%(P0.01).3 on the lesion side of 6-OHDA Parkinson's disease model rats. Injection of 0.01mM orexin-A under micropressure could significantly increase the discharge frequency of 12 dopaminergic neurons recorded in the substantia nigra compact band (12 of 21 dopaminergic neurons recorded in the substantia nigra compact band).From 4.1 鹵0.6Hz to 6.5 鹵1.0Hz, the average increase was 58.2 鹵11.8hz.Among the 12 GABA neurons recorded in the substantia nigra reticularis nigra, the firing frequency of 7 neurons was significantly increased from 19.6 鹵2.7Hz to 23.5 鹵3.8 Hz, and the average increase was 18.8 鹵3.0 Hz.In normal and 6-OHDA Parkinson's disease model rats, the excitatory effect of lorexin-A on the dopaminergic neurons in the substantia nigra compacta is stronger than that in the substantia nigra reticularis (P0.01 in normal rats and P0.05. 4 in Parkinson's disease model rats).Immunohistochemical staining showed that OX1 receptors were expressed in the substantia nigra of normal and Parkinson's disease rats.Conclusion: Orexin-A can increase the excitability of dopaminergic neurons in substantia nigra and GABA neurons in reticular zone.Endogenous OX1 receptors are involved in the regulation of excitability of dopaminergic neurons in the substantia nigra compact zone and GABA neurons in the reticular zone.In Parkinson's disease model rats, moorexin-A also increased the excitability of dopaminergic neurons in the substantia nigra compact zone and GABA neurons in the reticular zone of the substantia nigra.Both normal and Parkinson's disease model rats expressed OX1 receptors in substantia nigra.The results provide a theoretical and experimental basis for the further study of the role of substantia nigra orexin-A in the pathogenesis of Parkinson's disease.
【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】：2014
【分類號】：R742.5

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