本文關鍵詞： orexin-A 外側核 伏隔核 葡萄糖敏感神經元 胃運動　出處：《青島大學》2017年碩士論文　論文類型：學位論文

【摘要】：目的:闡明下丘腦外側核(LHA)-伏隔核(NAcc)orexin-A神經和功能通路構成,探討該通路對大鼠葡萄糖敏感神經元(GS)放電活動和胃運動的影響及潛在機制。方法:健康成年雄性Wistar大鼠隨機分為三部分:第一部分大鼠采用逆行追蹤技術結合免疫熒光組織化學染色法,觀察下丘腦外側核-伏隔核間是否存在orexin-A神經通路;第二部分大鼠采用單細胞外記錄放電活動,鑒定伏隔核葡萄糖敏感神經元,并且觀察伏隔核內微量注射orexin-A對葡萄糖敏感神經元放電活動的影響,以及電刺激下丘腦外側核后,伏隔核內orexin-A反應性葡萄糖敏感神經元放電活動的變化;第三部分大鼠通過在體胃運動研究,觀察伏隔核內微量注射不同濃度orexin-A對大鼠胃運動幅度和頻率的影響,以及電刺激下丘腦外側核后,大鼠胃運動的變化及機制。結果:逆行追蹤結合熒光免疫組織化學染色發(fā)現(xiàn),下丘腦外側核內有熒光金和orexin-A雙重標記的神經元。電生理研究顯示,在伏隔核內共記錄到62個葡萄糖敏感型神經元(GS),其中有42個GS呈現(xiàn)興奮效應(GS-E),20個GS呈現(xiàn)抑制效應(GS-I)。伏隔核微量注射orexin-A,在這42個GS-E中,有30個神經元(30/42,71.43%)放電頻率顯著升高(P0.05),10個神經元(10/42,23.81%)放電頻率顯著降低(P0.05)。在20個GS-I中,伏隔核微量注射orexin-A,其中有15個神經元(15/20,75%)放電活動呈現(xiàn)興奮效應(P0.05),有4個神經元(4/20,20%)放電活動呈現(xiàn)抑制效應(P0.05)。伏隔核預先注射orexin-A受體拮抗劑SB-334867,orexin-A對葡萄糖敏感神經元的興奮或抑制作用則被完全阻斷(P0.05)。電刺激下丘腦外側核,在伏隔核記錄到的39個orexin-A反應性GS-E神經元中,有28個神經元(28/39,71.79%)進一步呈現(xiàn)興奮效應(P0.05),7個神經元(7/39,17.94%)放電活動呈現(xiàn)抑制狀態(tài)(P0.05);在伏隔核記錄到的20個orexin-A反應性GS-I神經元,其中有14個神經元(14/20,70%)放電頻率顯著升高(P0.05),4個神經元(20.0%)放電活動顯著減弱(P0.05)。伏隔核預先注射SB-334867,再電刺激下丘腦外側核,伏隔核內orexin-A反應性葡萄糖敏感神經元的放電頻率均顯著下降(P0.05)。胃運動研究結果顯示,伏隔核內微量注射orexin-A,大鼠胃運動幅度和頻率顯著增加,并呈現(xiàn)顯著劑量依賴關系(P0.05),伏隔核預先微量注射SB-334867,可反轉該效應(P0.05)。電刺激下丘腦外側核,大鼠胃運動幅度和頻率顯著增強(P0.05)。同樣,伏隔核內微量注射SB-334867,再電刺激下丘腦外側核,電刺激導致的胃運動增強效應顯著減弱(P0.05)。結論:下丘腦外側核→伏隔核存在orexin-A神經和功能通路,該通路可能參與胃動力和能量代謝調控。
[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.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R338

【相似文獻】

相關期刊論文 前10條

1 趙琳;賈鯤鵬;史娟;苗乃周;王璐;;禁食不同時段下丘腦穹窿周區(qū)orexin-A的表達[J];中國組織化學與細胞化學雜志;2012年06期

2 葉峻;;樹，

本文編號：1442936