【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)控胃腸運動、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,并研究了該通路對胃擴(kuò)張(gastric distention,GD)敏感神經(jīng)元放電活動及胃運動的影響。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動實驗,觀察LHA-NTS ghrelin通路對嘔吐大鼠GD神經(jīng)元放電活動及胃運動的影響。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動障礙。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及胃運動的調(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)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R730.53

【參考文獻(xiàn)】

相關(guān)期刊論文 前9條

1 盧靈莉;張偉鋒;陳華林;;T淋巴細(xì)胞、血清炎癥因子與胃腸炎伴良性驚厥的相關(guān)性[J];兒科藥學(xué)雜志;2016年09期

2 陳彩萍;馮志奇;宋壯;陳超;溫小安;袁浩亮;;蛋白免疫印跡法同時檢測大、小分子蛋白的實驗條件改進(jìn)[J];現(xiàn)代生物醫(yī)學(xué)進(jìn)展;2016年24期

3 林敏;涂遠(yuǎn)榮;李旭;賴繁彩;陳劍鋒;邱明鏈;;大鼠前肢足墊皮膚交感神經(jīng)HRP逆行示蹤研究[J];中國醫(yī)藥科學(xué);2012年23期

4 高云;陳嘉昌;王偵;彭煥玉;李明;朱振宇;陳華云;;多重PCR點突變篩查技術(shù)在CYP2C19基因分型中的應(yīng)用[J];中華醫(yī)學(xué)雜志;2011年32期

5 汪靖超,李榮貴;重組TaqDNA聚合酶在大腸桿菌中的表達(dá)與純化[J];青島大學(xué)學(xué)報(工程技術(shù)版);2004年01期

6 黃祥瑞,陳娟,李曉萸,楊秀旭,吳慶麗,李德榮;西藏環(huán)狀病毒細(xì)胞生物學(xué)特性研究[J];軍事醫(yī)學(xué)科學(xué)院院刊;2000年03期

7 張瑩,徐文弟;酶聯(lián)免疫吸附測定法在醫(yī)學(xué)研究中的新進(jìn)展[J];哈爾濱醫(yī)科大學(xué)學(xué)報;2000年04期

8 戴玉琳，蘇　;同步ELISA試驗檢測8種抗核抗體[J];上海醫(yī)學(xué)檢驗雜志;1996年03期

9 趙樹仲，，張文;電生理技術(shù)發(fā)展歷程的啟示[J];醫(yī)學(xué)與哲學(xué);1995年03期

相關(guān)博士學(xué)位論文 前1條

1 公衍玲;下丘腦弓狀核—外側(cè)隔核ghrelin通路構(gòu)成及對胃運動調(diào)控研究[D];青島大學(xué);2013年

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