LHA-NTS ghrelin神经通路的构成及对呕吐大鼠胃运动的调控研究
发布时间：2018-07-21 21:20
【摘要】：目的:作为生长激素促分泌素受体(growth hormone secretagogue receptor,GHSR)的唯一内源性配体,ghrelin具有调控胃肠运动、调节膳食、促进生长激素分泌等作用。本课题拟建立大鼠呕吐模型,探讨下丘脑外侧区(lateral hypothalamic area,LHA)至孤束核(nucleus tractus solitarius,NTS)ghrelin神经通路的构成及改变,并研究了该通路对胃扩张(gastric distention,GD)敏感神经元放电活动及胃运动的影响。方法:采用神经元逆行追踪和荧光免疫组化法研究LHA-NTS ghrelin神经通路的构成;腹腔注射顺铂建立大鼠呕吐模型;采用ELISA、PCR和Western blot法测定大鼠ghrelin及其受体GHSR-1a的变化;进行细胞外放电记录及胃运动实验,观察LHA-NTS ghrelin通路对呕吐大鼠GD神经元放电活动及胃运动的影响。结果:在NTS内观察到FG阳性细胞、ghrelin阳性细胞及双染细胞,提示LHA-NTS存在ghrelin神经通路;与生理盐水组比较,顺铂组大鼠胃及LHA ghrelin浓度降低(P0.05)。LHA和NTS GHSR-1a表达升高(P0.05)。两组大鼠NTS内记录到GD敏感神经元,NTS内注射ghrelin,GD神经元放电频率显著增加(P0.01),预先给予[D-Lys-3]-GHRP-6,该作用被完全阻断。电刺激LHA,NTS内ghrelin兴奋的GD神经元放电频率显著增加(P0.01),NTS注射[D-Lys-3]-GHRP-6,该作用被部分阻断。大鼠腹腔注射顺铂后胃收缩频率及振幅降低(P0.01),提示顺铂可造成胃运动障碍。大鼠NTS内注射ghrelin后胃收缩振幅和频率增加(P0.05-0.01),此作用可被[D-Lys-3]-GHRP-6完全阻断。电刺激LHA可使大鼠胃收缩振幅和频率增加(P0.01),预先于NTS注射[D-Lys-3]-GHRP-6,此作用被部分阻断。在上述作用中,顺铂组大鼠的兴奋作用均弱于生理盐水组大鼠(P0.05)。结论:LHA-NTS存在ghrelin神经通路,且该通路参与呕吐大鼠胃肠道传入信号及胃运动的调控。LHA内ghrelin分泌减少可能导致顺铂处理大鼠ghrelin神经通路作用减弱。且LHA和NTS的GHSR-1a适应性上调表明外源性ghrelin可缓解顺铂导致的相关症状。
[Abstract]:Aim: as the only endogenous ligand of growth hormone receptor (growth hormone secretagogue receptor (GHSR), ghrelin can regulate gastrointestinal motility, diet and growth hormone secretion. The purpose of this study was to establish a rat model of vomiting, to investigate the formation and changes of the ghrelin pathway from the lateral hypothalamic area (lateral hypothalamic area) to the nucleus solitarius (NTS) of the solitary tract, and to study the effects of the pathway on the discharges and gastric motility of gastric distension GD sensitive neurons. Methods: the neuronal retrograde tracing and fluorescence immunohistochemistry were used to study the structure of LHA-NTS ghrelin pathway, the rat model of vomiting was established by intraperitoneal injection of cisplatin, the changes of ghrelin and its receptor GHSR-1a were measured by ELISA-PCR and Western blot. The effects of LHA-NTS ghrelin pathway on the discharge activity and gastric motility of GD neurons in vomiting rats were observed. Results: FG-positive cells and double staining cells were observed in NTS, indicating that there was a ghrelin neural pathway in LHA-NTS, and that in cisplatin group, the concentrations of ghrelin in stomach and LHA were decreased (P0.05), and the expression of ghrelin and GHSR-1a were increased (P0.05) in Cisplatin group. The discharge frequency of GD neurons was significantly increased (P0.01) and [D-Lys-3] -GHRP-6 was given in advance, which was completely blocked. The firing frequency of GD neurons excited by ghrelin was significantly increased after electrical stimulation (P0.01). [D-Lys-3] -GHRP-6 was injected with [D-Lys-3], and the effect was partially blocked. The gastric contraction frequency and amplitude decreased after intraperitoneal injection of cisplatin (P0.01), suggesting that cisplatin could cause gastric motility disorder. The amplitude and frequency of gastric contraction increased after ghrelin injection (P0.05-0.01), which could be completely blocked by [D-Lys-3] -GHRP-6. Electrical stimulation of LHA increased the amplitude and frequency of gastric contraction in rats (P0.01) and was partially blocked by injection of [D-Lys-3] -GHRP-6 into NTS. The excitatory effect of cisplatin group was weaker than that of normal saline group (P0.05). Conclusion there is a ghrelin neural pathway in 1: LHA-NTS, and this pathway is involved in the regulation of gastrointestinal afferent signal and gastric motility in vomit rats. The decrease of ghrelin secretion in LHA may lead to the decrease of ghrelin nerve pathway in rats treated with cisplatin. The up-regulation of GHSR-1a in LHA and NTS suggests that exogenous ghrelin can relieve the symptoms caused by cisplatin.
【学位授予单位】：青岛科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R730.53
本文编号：2136891
[Abstract]:Aim: as the only endogenous ligand of growth hormone receptor (growth hormone secretagogue receptor (GHSR), ghrelin can regulate gastrointestinal motility, diet and growth hormone secretion. The purpose of this study was to establish a rat model of vomiting, to investigate the formation and changes of the ghrelin pathway from the lateral hypothalamic area (lateral hypothalamic area) to the nucleus solitarius (NTS) of the solitary tract, and to study the effects of the pathway on the discharges and gastric motility of gastric distension GD sensitive neurons. Methods: the neuronal retrograde tracing and fluorescence immunohistochemistry were used to study the structure of LHA-NTS ghrelin pathway, the rat model of vomiting was established by intraperitoneal injection of cisplatin, the changes of ghrelin and its receptor GHSR-1a were measured by ELISA-PCR and Western blot. The effects of LHA-NTS ghrelin pathway on the discharge activity and gastric motility of GD neurons in vomiting rats were observed. Results: FG-positive cells and double staining cells were observed in NTS, indicating that there was a ghrelin neural pathway in LHA-NTS, and that in cisplatin group, the concentrations of ghrelin in stomach and LHA were decreased (P0.05), and the expression of ghrelin and GHSR-1a were increased (P0.05) in Cisplatin group. The discharge frequency of GD neurons was significantly increased (P0.01) and [D-Lys-3] -GHRP-6 was given in advance, which was completely blocked. The firing frequency of GD neurons excited by ghrelin was significantly increased after electrical stimulation (P0.01). [D-Lys-3] -GHRP-6 was injected with [D-Lys-3], and the effect was partially blocked. The gastric contraction frequency and amplitude decreased after intraperitoneal injection of cisplatin (P0.01), suggesting that cisplatin could cause gastric motility disorder. The amplitude and frequency of gastric contraction increased after ghrelin injection (P0.05-0.01), which could be completely blocked by [D-Lys-3] -GHRP-6. Electrical stimulation of LHA increased the amplitude and frequency of gastric contraction in rats (P0.01) and was partially blocked by injection of [D-Lys-3] -GHRP-6 into NTS. The excitatory effect of cisplatin group was weaker than that of normal saline group (P0.05). Conclusion there is a ghrelin neural pathway in 1: LHA-NTS, and this pathway is involved in the regulation of gastrointestinal afferent signal and gastric motility in vomit rats. The decrease of ghrelin secretion in LHA may lead to the decrease of ghrelin nerve pathway in rats treated with cisplatin. The up-regulation of GHSR-1a in LHA and NTS suggests that exogenous ghrelin can relieve the symptoms caused by cisplatin.
【学位授予单位】：青岛科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R730.53
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