【摘要】： 目的：研究胃動(dòng)素對(duì)大鼠弓狀核胃擴(kuò)張反應(yīng)神經(jīng)元放電活動(dòng)和胃運(yùn)動(dòng)的影響,探討弓狀核參與調(diào)控胃運(yùn)動(dòng)的機(jī)制。 方法：1.電生理實(shí)驗(yàn)方法：應(yīng)用四管玻璃微電極細(xì)胞外記錄麻醉大鼠弓狀核神經(jīng)元自發(fā)放電,用水囊充盈胃鑒別胃擴(kuò)張敏感神經(jīng)元,以壓力注射儀經(jīng)玻璃微電極,對(duì)核團(tuán)內(nèi)單個(gè)神經(jīng)元分別微量注射給予：(1)胃動(dòng)素、生理鹽水(對(duì)照組)(2)胃動(dòng)素、GM-109(胃動(dòng)素受體拮抗劑)觀察藥物對(duì)神經(jīng)元單位放電的影響。2.應(yīng)用核團(tuán)微量注射法,記錄清醒自由活動(dòng)大鼠胃運(yùn)動(dòng),觀察向大鼠弓狀核內(nèi)分別微量注射胃動(dòng)素和胃動(dòng)素受體拮抗劑GM-109后,對(duì)清醒自由活動(dòng)大鼠胃運(yùn)動(dòng)的變化。3.應(yīng)用熒光免疫組織化學(xué)方法觀察胃動(dòng)素免疫反應(yīng)性神經(jīng)元在弓狀核的表達(dá)。 結(jié)果：(1)電生理實(shí)驗(yàn)結(jié)果：在35只大鼠中,記錄到110個(gè)的神經(jīng)元為胃擴(kuò)張敏感性神經(jīng)元,其中55.6%呈GD興奮反應(yīng),為胃擴(kuò)張興奮性神經(jīng)元(GD-E)；44.4%呈GD抑制反應(yīng),為GD抑制性神經(jīng)元(GD-I)。胃擴(kuò)張后,GD-E神經(jīng)元放電頻率由3.18±1.01Hz增至5.62±1.22Hz(P0.01),而且GD-I神經(jīng)元放電頻率由3.21±0.68Hz降至1.24±0.31Hz(P0.01)。弓狀核內(nèi)壓力注射胃動(dòng)素后,70%的GD-E神經(jīng)元表現(xiàn)為興奮作用,17.5%表現(xiàn)為抑制作用,并且放電頻率由3.46±1.59Hz增至8.83±2.25Hz(P0.05)；而注射胃動(dòng)素后,65.6%的GD-I神經(jīng)元表現(xiàn)為興奮作用,放電頻率由3.34±0.63Hz增至6.17±0.97Hz(P0.05)。而這種由胃動(dòng)素誘導(dǎo)的興奮作用被胃動(dòng)素受體拮抗劑GM-109阻斷。 (2)胃運(yùn)動(dòng)實(shí)驗(yàn)：弓狀核內(nèi)微量注射胃動(dòng)素可增強(qiáng)清醒大鼠胃運(yùn)動(dòng),其反應(yīng)與注入的胃動(dòng)素濃度呈劑量依賴關(guān)系；弓狀核內(nèi)先單獨(dú)注入胃動(dòng)素受體拮抗劑(GM-109)可阻斷由胃動(dòng)素誘導(dǎo)的對(duì)胃運(yùn)動(dòng)的增強(qiáng)作用。 (3)熒光免疫組化實(shí)驗(yàn)：經(jīng)過胃擴(kuò)張后,弓狀核內(nèi)胃動(dòng)素免疫反應(yīng)性神經(jīng)元數(shù)量增多,由10.92±0.48 cells/mm2增至43.68±0.44 cells/mm2(P0.05). 結(jié)論：下丘腦弓狀核內(nèi)存在對(duì)來自胃機(jī)械感受傳入信號(hào)產(chǎn)生反應(yīng)的胃擴(kuò)張反應(yīng)性神經(jīng)元；胃動(dòng)素可增強(qiáng)大鼠弓狀核內(nèi)胃擴(kuò)張反應(yīng)性神經(jīng)元的電活動(dòng),且胃動(dòng)素誘導(dǎo)的這一興奮作用可被胃動(dòng)素受體拮抗劑GM-109所阻斷；中樞給予胃動(dòng)素后,可引起大鼠胃運(yùn)動(dòng)活動(dòng)增強(qiáng),且這一增強(qiáng)作用可被GM-109阻斷。通過胃擴(kuò)張這一機(jī)械感受刺激后,弓狀核內(nèi)胃動(dòng)素免疫反應(yīng)性神經(jīng)元數(shù)量增多。研究證實(shí)了弓狀核胃動(dòng)素神經(jīng)元接收來自胃感受器的外周軀體感覺傳入神經(jīng)的沖動(dòng),并通過某些下級(jí)核團(tuán)通路發(fā)揮胃運(yùn)動(dòng)的調(diào)節(jié)作用。該結(jié)果為弓狀核在胃動(dòng)素調(diào)控胃腸運(yùn)動(dòng)中的作用開闊了前景。
[Abstract]:Aim: to study the effects of motilin on gastric distension response neurons and gastric motility in rat arcuate nucleus, and to explore the mechanism of arcuate nucleus involved in regulating gastric motility. Method 1: 1. Methods: the spontaneous discharges of arcuate nucleus neurons in anesthetized rats were recorded by using four tubes of glass microelectrode. The gastric dilatation sensitive neurons were identified by filling the stomach with water sac. The nerve cells were treated with a pressure injector through glass microelectrode. (1) motilin, normal saline (control group) and (2) motilin GM-109 (motilin receptor antagonist) were used to observe the effect of motilin on the unit discharge of neurons. The gastric motility of conscious and free moving rats was recorded by micronucleus injection. The changes of motilin and motilin receptor antagonist GM-109 in arcuate nucleus of rats were observed. The expression of motilin immunoreactive neurons in the arcuate nucleus was observed by fluorescence immunohistochemical method. Results: (1) results of electrophysiological experiment: 110 neurons were recorded as gastric dilatation sensitive neurons in 35 rats. 55.6% of them showed GD excitatory response, and 44.4% of them showed GD inhibitory response to gastric dilatation excitatory neurons (GD-E). GD inhibitory neurons (GD-I). After gastric dilatation, the discharge frequency of GD-E neurons increased from 3.18 鹵1.01Hz to 5.62 鹵1.22Hz (P0.01), and the frequency of GD-I neurons decreased from 3.21 鹵0.68Hz to 1.24 鹵0.31Hz (P0.01). After motilin injection, 70% of the GD-E neurons in the arcuate nucleus showed an excitatory effect, and the discharge frequency increased from 3.46 鹵1.59 Hz to 8.83 鹵2.25 Hz (P0.05), while that of 65.6% of the GD-I neurons after injection of motilin increased from 3.34 鹵0.63 Hz to 6.17 鹵0.97 Hz (P0.05), and that of the GD-E neurons increased from 3.34 鹵0.63 Hz to 6.17 鹵0.97 Hz (P0.05), while that of the motilin group was increased from 3.34 鹵0.63 Hz to 6.17 鹵0.97 Hz (P0.05). The motilin induced excitatory effect was blocked by motilin receptor antagonist GM-109. (2) gastric motility experiment: motilin microinjection into arcuate nucleus enhanced gastric motility in conscious rats. There was a dose-dependent relationship between the response and the concentration of motilin injected. The motilin receptor antagonist (GM-109) alone injected into the arcuate nucleus could block the motilin induced enhancement of gastric motility. (3) fluorescence immunohistochemistry: after gastric dilatation, the motilin receptor antagonist (GM-109) was injected into the arcuate nucleus alone. The number of motilin immunoreactive neurons in arcuate nucleus increased from 10.92 鹵0.48 cells/mm2 to 43.68 鹵0.44 cells/mm2 (P0.05). Conclusion: there are gastric dilatation responsive neurons in the arcuate nucleus of the hypothalamus that respond to the afferent signals from the gastric machinery, and motilin can enhance the electrical activity of the gastric dilatation responsive neurons in the arcuate nucleus of rats. The stimulation induced by motilin could be blocked by motilin receptor antagonist GM-109, and the central administration of motilin could induce the increase of gastric motility in rats, which could be blocked by GM-109. The number of motilin immunoreactive neurons in the arcuate nucleus increased after the mechanical stimulation of gastric expansion. It is confirmed that motilin neurons in arcuate nucleus receive the impulses of peripheral somatosensory afferent nerves from gastric receptors and play a regulatory role in gastric motility through some lower nuclear pathways. The results show that the role of arcuate nucleus in motilin regulating gastrointestinal motility is promising.
【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】：2010
【分類號(hào)】：R333

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