本文選題：人工合成肽 + 鼻腔給藥��； 參考：《桂林醫(yī)學(xué)院》2012年碩士論文

【摘要】：目的：人工合成目的肽DSIP并分析純化,研究其促眠活性,并進(jìn)一步研究經(jīng)鼻腔給藥的促眠作用,同時(shí)探索毒副作用,從而為治療失眠癥提供一種人工合成因而不受來(lái)源限制、質(zhì)量易于控制,在失眠癥的治療中取得較好效果、給藥方便并免除不良副作用,可迅速推廣應(yīng)用的高效外源性促眠藥物。方法：利用生物信息學(xué)對(duì)已知結(jié)構(gòu)的天然存在睡眠肽進(jìn)行分析,選擇其α同分異構(gòu)體并采用全化學(xué)合成的固相多肽合成法合成DSIP,通過(guò)高效液相色譜和質(zhì)譜法進(jìn)行純化分析及鑒定,確定合成的多肽即為設(shè)計(jì)需要的目的肽并分析其純度。進(jìn)行分組動(dòng)物實(shí)驗(yàn)研究目的肽的促眠效果：用大鼠初步判定此人工合成的目的肽DSIP是否具有促眠作用；確認(rèn)其效用后第一步用新西蘭大耳白兔判定目的肽經(jīng)靜脈給藥是否如文獻(xiàn)報(bào)道的有誘導(dǎo)新西蘭大耳白兔睡眠的作用；第二步研究鼻腔給藥并比較不同劑量DSIP的促眠效果,初步探索出目的肽鼻腔給藥的適宜劑量。最后通過(guò)病理學(xué)實(shí)驗(yàn)檢測(cè)人工合成的睡眠肽DSIP在大中劑量中長(zhǎng)期給藥后對(duì)腦組織是否有毒性。結(jié)果：1.成功合成具有生物活性的睡眠肽DSIP,經(jīng)高效液相法純化后達(dá)到所需要的95%以上純度,質(zhì)譜法與HPLC法確認(rèn)合成的是目的肽；2.安裝新西蘭大耳白兔腦電極后兔子仍然存活狀況良好,給藥前后均成功引導(dǎo)出新西蘭大耳白兔的腦電圖,記錄數(shù)據(jù)得到有效分析；3.分析腦電圖并進(jìn)行統(tǒng)計(jì)學(xué)檢驗(yàn)后,鼻腔給藥組、靜脈給藥組新西蘭大耳白兔均與生理鹽水組新西蘭大耳白兔總睡眠時(shí)間、慢波睡眠時(shí)間統(tǒng)計(jì)學(xué)有顯著性差異,且給藥組慢波睡眠時(shí)間長(zhǎng)、總睡眠時(shí)間長(zhǎng)；4.分析腦電圖并進(jìn)行統(tǒng)計(jì)學(xué)檢驗(yàn)后,鼻腔給藥15nmol/kg組與30nmol/kg組及45nmol/kg組新西蘭大耳白兔總睡眠時(shí)間、慢波睡眠時(shí)間統(tǒng)計(jì)學(xué)有顯著性差異,鼻腔給藥30nmol/kg組、及45nmol/kg組新西蘭大耳白兔與靜注劑量30nmol/kg組新西蘭大耳白兔總睡眠時(shí)間、慢波睡眠時(shí)間統(tǒng)計(jì)學(xué)無(wú)顯著性差異；5.中長(zhǎng)期大中劑量DSIP靜脈給藥后光鏡下對(duì)照組和給藥組的腦組織切片經(jīng)HE染色經(jīng)肉眼觀察無(wú)明顯差異。結(jié)論：1.人工合成的睡眠肽DSIP通過(guò)鼻腔給藥有促眠作用；2.人工合成的睡眠肽DSIP通過(guò)鼻腔給藥可得到靜注相近的促眠作用,初步認(rèn)為30nmol/kg為新西蘭大耳白兔鼻腔給藥適宜劑量；3.人工合成的睡眠肽DSIP中長(zhǎng)期大中劑量給藥對(duì)腦組織細(xì)胞無(wú)明顯損傷。
[Abstract]:Objective: to synthesize the objective peptide DSIP, analyze and purify it, study its activity of promoting sleep, further study the effect of nasal administration, and explore the side effects, so as to provide an artificial synthesis for the treatment of insomnia, so as not to be limited by the source.The quality is easy to control, and good effect has been obtained in the treatment of insomnia. It is convenient to give drugs and avoid adverse side effects. It can be used quickly to popularize the highly effective exogenous sleep-promoting drugs.Methods: the natural sleep peptides of known structures were analyzed by bioinformatics.The 偽 -isomer was selected and synthesized by solid-phase polypeptide synthesis. The synthesized peptide was purified and identified by HPLC and MS. The synthesized peptide was determined to be the target peptide for design and its purity was analyzed.To study the effect of the objective peptide on promoting sleep: the rat was used to determine whether the synthetic peptide DSIP had the effect of promoting sleep.In the first step, New Zealand Big ear White Rabbit was used to determine whether intravenous administration of the target peptide could induce sleep in New Zealand Big ear White Rabbit as reported in the literature, and the second step was to study nasal administration and compare the effects of different doses of DSIP on sleep promotion in New Zealand Big ear White Rabbit.Objective: to explore the appropriate dose for nasal administration of objective peptide.Finally, the toxicity of synthetic sleep peptide DSIP to brain tissue was detected by pathological experiments.The result is 1: 1.The sleep peptide DSIPs with biological activity were successfully synthesized and purified by high performance liquid chromatography (HPLC) to achieve the desired purity of more than 95%. It was confirmed by mass spectrometry and HPLC that the synthesized peptide was the target peptide 2.The rabbit survived well after installing the brain electrode of New Zealand big ear white rabbit. The EEG of New Zealand big ear white rabbit was successfully guided before and after administration, and the recorded data were analyzed effectively.After electroencephalogram analysis and statistical test, there were significant differences in the total sleep time and slow wave sleep time between the nasal administration group and intravenous administration group and normal saline group.The slow wave sleep time was long and the total sleep time was 4.After analysis of EEG and statistical test, there were significant differences in total sleep time and slow wave sleep time between 15nmol/kg group, 30nmol/kg group and 45nmol/kg group.There was no significant difference in total sleep time and slow wave sleep time between 45nmol/kg group and 30nmol/kg group.There was no significant difference in brain sections between the control group and the administration group by HE staining under light microscope after medium and long term intravenous administration of DSIP.Conclusion 1.Synthetic sleep peptide DSIP can promote sleep by nasal administration.Synthetic sleep peptide DSIP can get similar sleep promoting effect by nasal administration. It is preliminarily considered that 30nmol/kg is the appropriate dose for nasal administration of New Zealand big ear white rabbits.The synthetic sleep peptide DSIP was given in medium and long term dose without obvious damage to brain tissue cells.
【學(xué)位授予單位】：桂林醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R363

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