本文選題：大鼠 + 重度磨耗�。� 參考：《第四軍醫(yī)大學(xué)》2014年碩士論文

【摘要】：研究背景： 重度磨耗（severe occlusal attrition，SOA）作為口腔臨床上常見的慢性疾病，是導(dǎo)致顳下頜關(guān)節(jié)紊亂病（temporomandibular joint disorder，TMD）的一個重要原因，常伴隨著疼痛、下頜運動異常、顳下頜關(guān)節(jié)（temporomandibular joint，TMJ）區(qū)彈響和雜音等癥狀，其中咀嚼肌和顳下頜關(guān)節(jié)區(qū)疼痛常為患者就診的首要原因。前期關(guān)于重度磨耗的基礎(chǔ)研究主要集中于口腔局部組織的反應(yīng)，研究證實重度磨耗會引起咀嚼肌和顳下頜關(guān)節(jié)發(fā)生適應(yīng)性改建，但是針對重度磨耗引起的口頜面部疼痛以及神經(jīng)系統(tǒng)相關(guān)反應(yīng)的研究較少。在其他口頜面部神經(jīng)病理性痛和炎性痛的模型中發(fā)現(xiàn)，傷害性刺激可引起外周感覺神經(jīng)中樞三叉神經(jīng)節(jié)（trigeminal ganglion，TG）內(nèi)降鈣素基因相關(guān)肽（calcitonin gene-related peptide，CGRP）表達增多，腦干及延髓三叉神經(jīng)脊束核（spinal nucleus of trigeminal nerve，SpⅤ）區(qū)域內(nèi)的細(xì)胞絲裂原活化蛋白激酶（mitogen-activated protein kinases，MAPK）通路激活，磷酸化p38（phospho-p38，p-p38）蛋白的表達水平上調(diào)。因此，本研究擬以疼痛作為研究重點，分析重度磨耗后口頜面部疼痛的閾值變化及神經(jīng)系統(tǒng)的相關(guān)反應(yīng)。 研究目的： 建立并評價重度磨耗的動物模型，觀察重度磨耗后大鼠咬肌機械壓痛敏感程度的變化，并利用相關(guān)實驗技術(shù)手段觀察三叉神經(jīng)節(jié)、腦干及延髓內(nèi)的相關(guān)神經(jīng)遞質(zhì)的變化，以分析重度磨耗后大鼠外周及初級中樞神經(jīng)系統(tǒng)的反應(yīng)。 研究方法： 1.采用外科顯微鏡下逐漸均勻降低大鼠上頜后牙牙冠的方法建立重度磨耗動物模型，觀察大鼠的表現(xiàn)型及體重變化，利用行為學(xué)方法檢測大鼠咬肌機械壓痛的敏感程度。 2.利用免疫熒光染色方法對重度磨耗后大鼠三叉神經(jīng)節(jié)內(nèi)CGRP表達水平的變化進行分析。 3.利用免疫熒光染色和蛋白免疫印跡方法對重度磨耗后大鼠腦干及延髓中p38MAPK表達水平的變化進行分析。 研究結(jié)果： 1.采用外科顯微鏡下逐漸均勻降低大鼠上頜后牙牙冠的方法成功建立了重度磨耗的動物模型，建模后大鼠表現(xiàn)型無明顯異常。3d時咬肌機械壓痛閾值顯著降低，持續(xù)到21d，第28d時基本恢復(fù)到基線水平。 2.重度磨耗可引起大鼠三叉神經(jīng)節(jié)上頜牙槽神經(jīng)神經(jīng)元相關(guān)區(qū)域內(nèi)的CGRP表達水平的升高。建模后的3d CGRP陽性神經(jīng)元的數(shù)量顯著增加，達到峰值，隨后3wCGRP陽性神經(jīng)元的數(shù)量有所減少但仍明顯高于對照組，第28d時基本恢復(fù)到正常水平。 3.重度磨耗后大鼠腦干及延髓三叉神經(jīng)脊束核相關(guān)區(qū)域內(nèi)p38MAPK通路被激活，建模后3d p-p38蛋白表達顯著上調(diào)，達到峰值后開始逐漸減少，激活狀態(tài)持續(xù)3w，第28d時p-p38蛋白的表達基本恢復(fù)到正常水平。 結(jié)論： 1.本實驗成功建立了重度磨耗的動物模型，行為學(xué)方法檢測發(fā)現(xiàn)大鼠重度磨耗后可以出現(xiàn)一過性的咬肌機械壓痛敏感性增高。 2.重度磨耗后大鼠三叉神經(jīng)節(jié)上頜牙槽神經(jīng)神經(jīng)元相關(guān)區(qū)域內(nèi)的CGRP表達水平一過性升高，，說明咬肌疼痛敏感性與三叉神經(jīng)節(jié)內(nèi)CGRP表達水平相關(guān)，進一步證實CGRP在此模型疼痛的發(fā)展過程中發(fā)揮了作用。 3.重度磨耗后大鼠腦干及延髓三叉神經(jīng)脊束核相關(guān)區(qū)域內(nèi)p-p38蛋白表達一過性增加，提示MAPK通路在重度磨耗建模后被激活，p38MAPK通路在此模型疼痛的調(diào)控中發(fā)揮著重要作用。
[Abstract]:Research background:
Severe occlusal attrition (SOA), as a common chronic disease in the oral cavity, is an important cause of the temporomandibular joint disorder (temporomandibular joint disorder, TMD), often accompanied by pain, abnormal mandibular movement, ringing and murmurs in the temporomandibular joint (temporomandibular joint, TMJ) area, including Tsui. The primary cause of pain in the masticatory and temporomandibular joint areas is often the primary cause of the patient's treatment. The basic research on severe abrasion is mainly focused on the response of the local tissue in the early stage. The study confirms that the severe attrition will cause the adaptation to the masticatory muscles and the temporomandibular joint, but the pain and the nerve system caused by severe wear and maxillofacial pain and the nervous system There are few studies on the related reactions. In the models of other oral and maxillofacial neuropathic pain and inflammatory pain, it is found that nociceptive stimulation can cause the increase in the expression of calcitonin gene-related peptide (CGRP) in the trigeminal ganglion (TG) and the trigeminal trigeminal spinal tract of the brain and medulla. The cell mitogen activated protein kinase (mitogen-activated protein kinases, MAPK) pathway in the nucleus (spinal nucleus of trigeminal nerve, Sp V) is activated and the expression level of phosphorylation p38 (phospho-p38) protein is up-regulated. Therefore, this study intends to focus on pain as a key point to analyze the threshold of pain in the oral and maxillofacial region after severe attrition. Changes in response to the nervous system.
The purpose of the study is:
To establish and evaluate the animal model of severe abrasion, observe the changes in the sensitivity of the masseter mechanical tenderness in the rats after severe abrasion, and observe the changes of the related neurotransmitters in the trigeminal ganglia, the brainstem and the medulla with relevant experimental techniques, in order to analyze the response of the peripheral and primary central nervous system in the severe attrition rats.
Research methods:
1. using the surgical microscope to gradually reduce the crown of the maxillary posterior teeth, a severe wear animal model was established to observe the changes of the phenotype and weight of the rats, and the sensitivity of the mechanical tenderness of the masseter muscle in rats was detected by the behavior method.
2. the expression of CGRP in trigeminal ganglion of severe wear rats was analyzed by immunofluorescence staining.
3. the expression of p38MAPK in brainstem and medulla oblongata was analyzed by immunofluorescence staining and Western blotting.
The results of the study:
1. the animal model of severe attrition was successfully established by the method of gradual uniform reduction of the maxillary posterior teeth crown under the surgical microscope. The threshold of the mechanical tenderness of the masseter muscle was significantly reduced when the model of the model had no obvious abnormal.3d. It continued to 21d, and the baseline was basically restored to the baseline at 28d.
2. severe attrition could increase the level of CGRP expression in the maxillary alveolar nerve neurons in the rat trigeminal ganglia. The number of 3D CGRP positive neurons after modeling increased significantly and reached the peak. Then the number of 3wCGRP positive neurons decreased but was still significantly higher than that of the control group. At the time of 28d, the number of positive neurons recovered to the normal level.
3. the p38MAPK pathway in the brain stem and the nucleus of the spinal trigeminal nucleus in the rats after severe abrasion was activated. After modeling, the expression of 3D p-p38 protein increased significantly. After the peak, the activation state gradually decreased and the activation state continued to be 3W. The expression of p-p38 protein at 28d was basically restored to normal level at the time of 28d.
Conclusion:
1. the animal models of severe attrition were successfully established in this experiment. The behavior method test found that the sensitivity of the mechanical tenderness of the masseter muscle increased after the severe attrition of the rats.
2. the level of CGRP expression in the maxillary alveolar nerve neurons in the trigeminal ganglion neurons of the rats after severe attrition increased, which indicates that the sensitivity of the masseter muscle pain is related to the level of CGRP expression in the trigeminal ganglia, and further confirms the role of CGRP in the development of this model.
3. the expression of p-p38 protein in the brain stem and the nucleus of the spinal trigeminal nucleus in the rats after severe attrition increased, suggesting that the MAPK pathway was activated after the modeling of severe attrition, and the p38MAPK pathway played an important role in the regulation of the model pain.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R781

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