本文選題：側(cè)腦室注射 + 痛覺(jué)過(guò)敏　； 參考：《泰山醫(yī)學(xué)院》2007年碩士論文

【摘要】： 目的以輻射熱刺激甩尾法為疼痛模型，旨在探討apelin在大鼠中樞痛覺(jué)調(diào)制中的作用及可能的作用機(jī)制。 方法 1．痛閾測(cè)定：雄性SD大鼠，以輻射熱引起大鼠甩尾，以大鼠甩尾反應(yīng)的潛伏期(tail-flick latency，TFL)作為痛反應(yīng)指標(biāo)。分別向側(cè)腦室微量注射生理鹽水(NS)、嗎啡(Morphine)和apelin，觀察注藥后60 min內(nèi)大鼠痛閾的變化情況。 2．腦內(nèi)nNOS表達(dá)的檢測(cè)：雄性SD大鼠，，隨機(jī)分為NS組、apelin組，采用免疫熒光方法在激光共聚焦顯微鏡下觀察給藥后30 min大鼠尾核、海馬及皮層nNOS表達(dá)的變化。 3．血清和腦內(nèi)NO、總NOS測(cè)定：雄性SD大鼠，分別向大鼠側(cè)腦室微量注射N(xiāo)S和apelin，給藥后30 min斷頭取血取腦，應(yīng)用硝酸還原酶法測(cè)定血清及腦內(nèi)NO和總NOS含量的變化。 4．血漿和腦內(nèi)cAMP和cGMP測(cè)定：雄性SD大鼠，分別向大鼠側(cè)腦室微量注射N(xiāo)S和apelin，給藥后30 min斷頭取血取腦，應(yīng)用放射免疫方法測(cè)定大鼠血漿及腦內(nèi)cAMP和cGMP含量的變化。 結(jié)果 1．側(cè)腦室微量注射apelin后10～30 min大鼠痛閾與NS對(duì)照組相比顯著降低，差異有統(tǒng)計(jì)學(xué)意義(P＜0.01)，30 min后大鼠痛閾逐漸回升，至60 min時(shí)大鼠痛閾仍低正常水平。 2．側(cè)腦室注射apelin后，激光共聚焦顯微鏡觀察到在關(guān)注腦區(qū)nNOS表達(dá)均發(fā)生變化。FITC標(biāo)記的nNOS呈現(xiàn)綠色熒光顆粒分布于細(xì)胞胞漿中，為典型的細(xì)胞質(zhì)分布；同時(shí)PI染色，細(xì)胞核為紅色熒光。整個(gè)圖像細(xì)胞形態(tài)完整，細(xì)胞核與細(xì)胞質(zhì)的熒光色彩標(biāo)識(shí)清晰可見(jiàn)。并且尾核及海馬nNOS神經(jīng)元較NS對(duì)照組表達(dá)明顯降低，差異有統(tǒng)計(jì)學(xué)意義(P＜0.05～0.01)。 3．側(cè)腦室微量注射apelin后，大鼠血清、尾核及海馬中NO和NOS含量明顯降低，與NS對(duì)照組比較，差異有統(tǒng)計(jì)學(xué)意義(P＜0.05～0.01)。 4．側(cè)腦室微量注射apelin后，大鼠血漿、尾核、海馬中cAMP含量升高，cGMP含量降低，與NS對(duì)照組比較，差異有統(tǒng)計(jì)學(xué)意義(P＜0.05～0.01)。 結(jié)論 1．側(cè)腦室微量注射apelin顯著降低大鼠痛閾，使大鼠痛覺(jué)敏感化。 2．Apelin的這種痛覺(jué)敏感化作用至少部分是通過(guò)尾核、海馬等核團(tuán)的調(diào)控作用實(shí)現(xiàn)的。 3．側(cè)腦室微量注射apelin明顯提高大鼠尾核、海馬及血漿中cAMP水平，降低大鼠尾核、海馬中NO和NOS及血清中NO的含量，同時(shí)使大鼠尾核、海馬及血漿中cGMP含量降低，提示apelin的痛覺(jué)敏感化作用可能與apelin-cAMP及apelin-NO-cGMP途徑相關(guān)。 本研究從行為學(xué)、組織和分子水平探討了apelin在大鼠中樞痛覺(jué)調(diào)制中的作用及其機(jī)制，為中樞神經(jīng)系統(tǒng)內(nèi)痛和鎮(zhèn)痛的研究提供了新的實(shí)驗(yàn)資料，對(duì)于進(jìn)一步探討痛覺(jué)調(diào)制的機(jī)制以及新藥物的開(kāi)發(fā)都有著重要的意義。
[Abstract]:Objective to investigate the role and possible mechanism of apelin in central pain modulation in rats. Method 1. Pain threshold measurement: male SD rats were induced tail flick by radiation heat, and tail flick latency TFL of tail flick reaction was used as the index of pain response. The changes of pain threshold were observed within 60 min after injection of NS, Morphine and apelin into lateral ventricle. 2. Detection of nNOS expression in brain: male SD rats were randomly divided into NS group and apelin group. The changes of nNOS expression in caudate nucleus, hippocampus and cortex of rats 30 min after administration were observed by immunofluorescence under confocal laser microscope. 3. Determination of no and total NOS in serum and brain: male Sprague-Dawley rats were microinjected with NS and apelin into lateral ventricle respectively. The blood and brain were taken 30 min after administration of the drug, and the changes of no and total NOS in serum and brain were measured by nitrate reductase method. 4. Determination of cAMP and cGMP in plasma and brain: male Sprague-Dawley rats were microinjected with NS and apelin into the lateral ventricle respectively. The blood samples were taken 30 min after administration of the drug. The contents of cAMP and cGMP in plasma and brain were measured by radioimmunoassay. Result 1. The pain threshold of rats at 1030 min after microinjection of apelin into the lateral ventricle was significantly lower than that of the NS control group (P < 0.01). After 30 min, the pain threshold of the rats gradually increased, and the pain threshold of the rats was still lower than that of the control group at 60 min. 2. After intracerebroventricular injection of apelin, it was observed by laser confocal microscopy that the expression of nNOS in the cerebral area of concern changed. FITC-labeled nNOS was distributed in the cytoplasm of the cells, which was typical cytoplasmic distribution and Pi staining. The nucleus is red fluorescent. The morphology of the whole image cell was complete, and the fluorescent color identification of the nucleus and cytoplasm was clearly visible. The expression of nNOS in caudate nucleus and hippocampus was significantly lower than that in NS control group (P < 0.05). 3. The contents of no and NOS in serum, caudate nucleus and hippocampus of rats were significantly decreased after microinjection of apelin into lateral ventricle, and the difference was statistically significant compared with NS control group (P < 0.05). 4. After microinjection of apelin into the lateral ventricle, the content of cAMP in plasma, caudate nucleus and hippocampus of rats increased and decreased, compared with NS control group, the difference was statistically significant (P < 0.05). Conclusion 1. Microinjection of apelin into lateral ventricle significantly reduced the pain threshold and sensitized the pain in rats. The pain sensitivity of 2.Apelin is at least partly mediated by the regulation of caudate and hippocampal nuclei. 3. Intracerebroventricular microinjection of apelin significantly increased the level of cAMP in caudate nucleus, hippocampus and plasma, decreased the content of no and NOS in caudate nucleus, hippocampus and serum, and decreased the content of cGMP in caudate nucleus, hippocampus and plasma of rats. The results suggest that the pain sensitivity of apelin may be related to the apelin-cAMP and apelin-NO-cGMP pathway. This study explored the role and mechanism of apelin in central pain modulation in rats at the behavioral, tissue and molecular levels, and provided new experimental data for the study of pain and analgesia in the central nervous system. It is of great significance to further explore the mechanism of pain modulation and the development of new drugs.
【學(xué)位授予單位】：泰山醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類(lèi)號(hào)】：R338

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