胃动素对大鼠弓状核胃扩张反应神经元放电活动和胃运动的影响
发布时间：2018-07-17 05:06
【摘要】： 目的：研究胃动素对大鼠弓状核胃扩张反应神经元放电活动和胃运动的影响,探讨弓状核参与调控胃运动的机制。 方法：1.电生理实验方法：应用四管玻璃微电极细胞外记录麻醉大鼠弓状核神经元自发放电,用水囊充盈胃鉴别胃扩张敏感神经元,以压力注射仪经玻璃微电极,对核团内单个神经元分别微量注射给予：(1)胃动素、生理盐水(对照组)(2)胃动素、GM-109(胃动素受体拮抗剂)观察药物对神经元单位放电的影响。2.应用核团微量注射法,记录清醒自由活动大鼠胃运动,观察向大鼠弓状核内分别微量注射胃动素和胃动素受体拮抗剂GM-109后,对清醒自由活动大鼠胃运动的变化。3.应用荧光免疫组织化学方法观察胃动素免疫反应性神经元在弓状核的表达。 结果：(1)电生理实验结果：在35只大鼠中,记录到110个的神经元为胃扩张敏感性神经元,其中55.6%呈GD兴奋反应,为胃扩张兴奋性神经元(GD-E)；44.4%呈GD抑制反应,为GD抑制性神经元(GD-I)。胃扩张后,GD-E神经元放电频率由3.18±1.01Hz增至5.62±1.22Hz(P0.01),而且GD-I神经元放电频率由3.21±0.68Hz降至1.24±0.31Hz(P0.01)。弓状核内压力注射胃动素后,70%的GD-E神经元表现为兴奋作用,17.5%表现为抑制作用,并且放电频率由3.46±1.59Hz增至8.83±2.25Hz(P0.05)；而注射胃动素后,65.6%的GD-I神经元表现为兴奋作用,放电频率由3.34±0.63Hz增至6.17±0.97Hz(P0.05)。而这种由胃动素诱导的兴奋作用被胃动素受体拮抗剂GM-109阻断。 (2)胃运动实验：弓状核内微量注射胃动素可增强清醒大鼠胃运动,其反应与注入的胃动素浓度呈剂量依赖关系；弓状核内先单独注入胃动素受体拮抗剂(GM-109)可阻断由胃动素诱导的对胃运动的增强作用。 (3)荧光免疫组化实验：经过胃扩张后,弓状核内胃动素免疫反应性神经元数量增多,由10.92±0.48 cells/mm2增至43.68±0.44 cells/mm2(P0.05). 结论：下丘脑弓状核内存在对来自胃机械感受传入信号产生反应的胃扩张反应性神经元；胃动素可增强大鼠弓状核内胃扩张反应性神经元的电活动,且胃动素诱导的这一兴奋作用可被胃动素受体拮抗剂GM-109所阻断；中枢给予胃动素后,可引起大鼠胃运动活动增强,且这一增强作用可被GM-109阻断。通过胃扩张这一机械感受刺激后,弓状核内胃动素免疫反应性神经元数量增多。研究证实了弓状核胃动素神经元接收来自胃感受器的外周躯体感觉传入神经的冲动,并通过某些下级核团通路发挥胃运动的调节作用。该结果为弓状核在胃动素调控胃肠运动中的作用开阔了前景。
[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 卤1.01Hz to 5.62 卤1.22Hz (P0.01), and the frequency of GD-I neurons decreased from 3.21 卤0.68Hz to 1.24 卤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 卤1.59 Hz to 8.83 卤2.25 Hz (P0.05), while that of 65.6% of the GD-I neurons after injection of motilin increased from 3.34 卤0.63 Hz to 6.17 卤0.97 Hz (P0.05), and that of the GD-E neurons increased from 3.34 卤0.63 Hz to 6.17 卤0.97 Hz (P0.05), while that of the motilin group was increased from 3.34 卤0.63 Hz to 6.17 卤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 卤0.48 cells/mm2 to 43.68 卤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.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R333
[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 卤1.01Hz to 5.62 卤1.22Hz (P0.01), and the frequency of GD-I neurons decreased from 3.21 卤0.68Hz to 1.24 卤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 卤1.59 Hz to 8.83 卤2.25 Hz (P0.05), while that of 65.6% of the GD-I neurons after injection of motilin increased from 3.34 卤0.63 Hz to 6.17 卤0.97 Hz (P0.05), and that of the GD-E neurons increased from 3.34 卤0.63 Hz to 6.17 卤0.97 Hz (P0.05), while that of the motilin group was increased from 3.34 卤0.63 Hz to 6.17 卤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 卤0.48 cells/mm2 to 43.68 卤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.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R333
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