【摘要】：目的:作為生長激素促分泌素受體(growth hormone secretagogue receptor,GHSR)的唯一內(nèi)源性配體,ghrelin具有調(diào)控胃腸運(yùn)動(dòng)、調(diào)節(jié)膳食、促進(jìn)生長激素分泌等作用。本課題擬建立大鼠嘔吐模型,探討下丘腦外側(cè)區(qū)(lateral hypothalamic area,LHA)至孤束核(nucleus tractus solitarius,NTS)ghrelin神經(jīng)通路的構(gòu)成及改變,并研究了該通路對(duì)胃擴(kuò)張(gastric distention,GD)敏感神經(jīng)元放電活動(dòng)及胃運(yùn)動(dòng)的影響。方法:采用神經(jīng)元逆行追蹤和熒光免疫組化法研究LHA-NTS ghrelin神經(jīng)通路的構(gòu)成;腹腔注射順鉑建立大鼠嘔吐模型;采用ELISA、PCR和Western blot法測(cè)定大鼠ghrelin及其受體GHSR-1a的變化;進(jìn)行細(xì)胞外放電記錄及胃運(yùn)動(dòng)實(shí)驗(yàn),觀察LHA-NTS ghrelin通路對(duì)嘔吐大鼠GD神經(jīng)元放電活動(dòng)及胃運(yùn)動(dòng)的影響。結(jié)果:在NTS內(nèi)觀察到FG陽性細(xì)胞、ghrelin陽性細(xì)胞及雙染細(xì)胞,提示LHA-NTS存在ghrelin神經(jīng)通路;與生理鹽水組比較,順鉑組大鼠胃及LHA ghrelin濃度降低(P0.05)。LHA和NTS GHSR-1a表達(dá)升高(P0.05)。兩組大鼠NTS內(nèi)記錄到GD敏感神經(jīng)元,NTS內(nèi)注射ghrelin,GD神經(jīng)元放電頻率顯著增加(P0.01),預(yù)先給予[D-Lys-3]-GHRP-6,該作用被完全阻斷。電刺激LHA,NTS內(nèi)ghrelin興奮的GD神經(jīng)元放電頻率顯著增加(P0.01),NTS注射[D-Lys-3]-GHRP-6,該作用被部分阻斷。大鼠腹腔注射順鉑后胃收縮頻率及振幅降低(P0.01),提示順鉑可造成胃運(yùn)動(dòng)障礙。大鼠NTS內(nèi)注射ghrelin后胃收縮振幅和頻率增加(P0.05-0.01),此作用可被[D-Lys-3]-GHRP-6完全阻斷。電刺激LHA可使大鼠胃收縮振幅和頻率增加(P0.01),預(yù)先于NTS注射[D-Lys-3]-GHRP-6,此作用被部分阻斷。在上述作用中,順鉑組大鼠的興奮作用均弱于生理鹽水組大鼠(P0.05)。結(jié)論:LHA-NTS存在ghrelin神經(jīng)通路,且該通路參與嘔吐大鼠胃腸道傳入信號(hào)及胃運(yùn)動(dòng)的調(diào)控。LHA內(nèi)ghrelin分泌減少可能導(dǎo)致順鉑處理大鼠ghrelin神經(jīng)通路作用減弱。且LHA和NTS的GHSR-1a適應(yīng)性上調(diào)表明外源性ghrelin可緩解順鉑導(dǎo)致的相關(guān)癥狀。
[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.
【學(xué)位授予單位】：青島科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R730.53

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