本文關(guān)鍵詞： orexin-A 外側(cè)核 伏隔核 葡萄糖敏感神經(jīng)元 胃運(yùn)動(dòng)　出處：《青島大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:闡明下丘腦外側(cè)核(LHA)-伏隔核(NAcc)orexin-A神經(jīng)和功能通路構(gòu)成,探討該通路對(duì)大鼠葡萄糖敏感神經(jīng)元(GS)放電活動(dòng)和胃運(yùn)動(dòng)的影響及潛在機(jī)制。方法:健康成年雄性Wistar大鼠隨機(jī)分為三部分:第一部分大鼠采用逆行追蹤技術(shù)結(jié)合免疫熒光組織化學(xué)染色法,觀察下丘腦外側(cè)核-伏隔核間是否存在orexin-A神經(jīng)通路;第二部分大鼠采用單細(xì)胞外記錄放電活動(dòng),鑒定伏隔核葡萄糖敏感神經(jīng)元,并且觀察伏隔核內(nèi)微量注射orexin-A對(duì)葡萄糖敏感神經(jīng)元放電活動(dòng)的影響,以及電刺激下丘腦外側(cè)核后,伏隔核內(nèi)orexin-A反應(yīng)性葡萄糖敏感神經(jīng)元放電活動(dòng)的變化;第三部分大鼠通過(guò)在體胃運(yùn)動(dòng)研究,觀察伏隔核內(nèi)微量注射不同濃度orexin-A對(duì)大鼠胃運(yùn)動(dòng)幅度和頻率的影響,以及電刺激下丘腦外側(cè)核后,大鼠胃運(yùn)動(dòng)的變化及機(jī)制。結(jié)果:逆行追蹤結(jié)合熒光免疫組織化學(xué)染色發(fā)現(xiàn),下丘腦外側(cè)核內(nèi)有熒光金和orexin-A雙重標(biāo)記的神經(jīng)元。電生理研究顯示,在伏隔核內(nèi)共記錄到62個(gè)葡萄糖敏感型神經(jīng)元(GS),其中有42個(gè)GS呈現(xiàn)興奮效應(yīng)(GS-E),20個(gè)GS呈現(xiàn)抑制效應(yīng)(GS-I)。伏隔核微量注射orexin-A,在這42個(gè)GS-E中,有30個(gè)神經(jīng)元(30/42,71.43%)放電頻率顯著升高(P0.05),10個(gè)神經(jīng)元(10/42,23.81%)放電頻率顯著降低(P0.05)。在20個(gè)GS-I中,伏隔核微量注射orexin-A,其中有15個(gè)神經(jīng)元(15/20,75%)放電活動(dòng)呈現(xiàn)興奮效應(yīng)(P0.05),有4個(gè)神經(jīng)元(4/20,20%)放電活動(dòng)呈現(xiàn)抑制效應(yīng)(P0.05)。伏隔核預(yù)先注射orexin-A受體拮抗劑SB-334867,orexin-A對(duì)葡萄糖敏感神經(jīng)元的興奮或抑制作用則被完全阻斷(P0.05)。電刺激下丘腦外側(cè)核,在伏隔核記錄到的39個(gè)orexin-A反應(yīng)性GS-E神經(jīng)元中,有28個(gè)神經(jīng)元(28/39,71.79%)進(jìn)一步呈現(xiàn)興奮效應(yīng)(P0.05),7個(gè)神經(jīng)元(7/39,17.94%)放電活動(dòng)呈現(xiàn)抑制狀態(tài)(P0.05);在伏隔核記錄到的20個(gè)orexin-A反應(yīng)性GS-I神經(jīng)元,其中有14個(gè)神經(jīng)元(14/20,70%)放電頻率顯著升高(P0.05),4個(gè)神經(jīng)元(20.0%)放電活動(dòng)顯著減弱(P0.05)。伏隔核預(yù)先注射SB-334867,再電刺激下丘腦外側(cè)核,伏隔核內(nèi)orexin-A反應(yīng)性葡萄糖敏感神經(jīng)元的放電頻率均顯著下降(P0.05)。胃運(yùn)動(dòng)研究結(jié)果顯示,伏隔核內(nèi)微量注射orexin-A,大鼠胃運(yùn)動(dòng)幅度和頻率顯著增加,并呈現(xiàn)顯著劑量依賴關(guān)系(P0.05),伏隔核預(yù)先微量注射SB-334867,可反轉(zhuǎn)該效應(yīng)(P0.05)。電刺激下丘腦外側(cè)核,大鼠胃運(yùn)動(dòng)幅度和頻率顯著增強(qiáng)(P0.05)。同樣,伏隔核內(nèi)微量注射SB-334867,再電刺激下丘腦外側(cè)核,電刺激導(dǎo)致的胃運(yùn)動(dòng)增強(qiáng)效應(yīng)顯著減弱(P0.05)。結(jié)論:下丘腦外側(cè)核→伏隔核存在orexin-A神經(jīng)和功能通路,該通路可能參與胃動(dòng)力和能量代謝調(diào)控。
[Abstract]:Objective: to elucidate the structure of NAccorexin-A nerve and functional pathway in LHAA-accumbent nucleus of lateral hypothalamic nucleus. Study on the effects of this pathway on glucose sensitive neurons in rats. Methods: healthy adult male Wistar rats were randomly divided into three parts: the first part used retrograde tracing technique combined with immunofluorescence histochemical staining. The orexin-A nerve pathway was observed between the lateral hypothalamic nucleus and nucleus accumbens. In the second part, the glucose sensitive neurons in nucleus accumbens were identified by single cell recording, and the effect of microinjection of orexin-A into nucleus accumbens on the activity of glucose sensitive neurons was observed. And the changes of orexin-A responsive glucose-sensitive neurons in nucleus accumbens after stimulation of lateral hypothalamic nucleus. The third part was to observe the effect of microinjection of different concentrations of orexin-A into nucleus accumbens on the amplitude and frequency of gastric motility and the stimulation of lateral hypothalamic nucleus by in vivo gastric motility in rats. Results: retrograde tracing combined with fluorescent immunohistochemical staining revealed the changes of gastric motility in rats. In the lateral hypothalamic nucleus, there were fluorescent gold and orexin-A labeled neurons. Electrophysiological studies showed that 62 glucose-sensitive neurons were recorded in the nucleus accumbens. Among them, 42 GS showed excitatory effect and 20 GS showed inhibitory effect. Orexin-A was microinjected into nucleus accumbens in 42 GS-E. There were 30 / 42 / 71.43 neurons with a significantly higher discharge frequency (P0.05 / 42) and 10 / 42 / 10 / 10 / 10 of 10 neurons. In 20 GS-I, orexin-A was microinjected into nucleus accumbens, of which 15 neurons were 15 / 20. The excitatory effect of firing activity was P0.05, with 4 neurons in 4 / 20. The discharges showed inhibitory effect (P 0.05). The nucleus accumbens was preinjected with orexin-A receptor antagonist SB-334867. The stimulation or inhibition of orexin-A on glucose-sensitive neurons was completely blocked by P0.05and electrical stimulation of the lateral hypothalamic nucleus. Of the 39 orexin-A reactive GS-E neurons recorded in nucleus accumbens, 28 / 39 / 71.79 neurons showed a further excitatory effect (P0.05). The 7 / 7 / 39 ~ 17.94 of the neurons showed a state of inhibition (P0.05). Of the 20 orexin-A reactive GS-I neurons recorded in nucleus accumbens, 14 / 20 / 20 ~ (70) had a significantly higher discharge frequency (P _ (0.05)). The discharge activity of 4 neurons decreased significantly (P 0.05). SB-334867 was injected into the nucleus accumbens before stimulation of the lateral hypothalamic nucleus. The discharge frequency of orexin-A responsive glucose-sensitive neurons in nucleus accumbens decreased significantly (P 0.05). Gastric motility study showed that orexin-A was microinjected into nucleus accumbens. The amplitude and frequency of gastric motility were increased significantly in a dose-dependent manner. SB-334867 was microinjected into nucleus accumbens in a dose-dependent manner. The amplitude and frequency of gastric motility in rats were significantly increased by P0.05. Similarly, SB-334867 was microinjected into nucleus accumbens. When the lateral hypothalamic nucleus was stimulated by electrical stimulation, the enhancement of gastric motility induced by electrical stimulation was significantly weakened (P 0.05). Conclusion: the lateral hypothalamic nucleus can be induced by the stimulation of the lateral hypothalamic nucleus. 鈫扵he nucleus accumbens has orexin-A neural and functional pathway, which may be involved in the regulation of gastric motility and energy metabolism.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R338

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