本文選題：電壓門控鈉離子通道 + Nav1.3��； 參考：《安徽醫(yī)科大學》2014年碩士論文

【摘要】：目的電壓門控鈉通道亞型Nav1.3屬于河豚毒素敏感型（TTX-S）鈉通道，Nav1.3主要表達于嚙齒動物胚胎腦組織，可快速激活和快速失活Na+電流，，對動作電位有顯著調(diào)節(jié)作用，在炎癥刺激和神經(jīng)元損傷后重新表達并出現(xiàn)異位。牙髓組織的感覺功能由疼痛感受器來支配，特定的神經(jīng)纖維構(gòu)成疼痛感受器。正常情況下，適宜外界刺激不會引起牙髓組織的不適反應(yīng)，但如繼發(fā)牙髓炎等牙髓炎性病變時，牙髓組織中的疼痛感受器會產(chǎn)生痛覺反應(yīng)。本實驗通過構(gòu)建實驗性大鼠牙髓炎模型，對比分析鈉通道Nav1.3在正常大鼠牙髓組織及輕度、中度、重度牙髓炎大鼠牙髓組織中表達的不同，探討鈉通道Nav1.3表達量與牙髓炎所致炎性疼痛之間的關(guān)系，有助于臨床上調(diào)整牙痛的治療靶點，有助于尋找優(yōu)于傳統(tǒng)局部麻醉劑的有效緩解牙痛的新方法。 方法將16只大鼠隨機分為對照組、實驗1d組、3d組、5d組，每組4只。大腸桿菌內(nèi)毒素(LPS)誘導(dǎo)大鼠牙髓炎后，在傷后1、3、5d處死，通過對構(gòu)造模型后對大鼠的動物行為學觀察，通過HE染色檢測對照組及實驗1d、3d、5d組牙髓組織的病理狀況，利用反轉(zhuǎn)錄聚合酶鏈（RT-PCR）法檢測對照組及實驗1d、3d、5d組牙髓組織中TNF-α mRNA水平的表達量，判斷牙髓炎嚴重程度與炎性疼痛之間的關(guān)系；利用免疫組織化學技術(shù)檢測Nav1.3在正常大鼠及牙髓炎大鼠牙髓組織中的定性表達；Western blot方法檢測并比較Nav1.3在正常大鼠及牙髓炎大鼠牙髓組織中的蛋白相對表達量。 結(jié)果1.動物行為學觀察結(jié)果表明，正常組大鼠飲食二便正常，精神狀態(tài)較佳；實驗組大鼠在造模后出現(xiàn)反復(fù)舔傷口，不敢咬食物，煩躁不安，尖叫不止，體重降低等表現(xiàn)；2.HE染色結(jié)果表明：正常組的組織結(jié)構(gòu)正常，無病理性改變；隨著牙髓炎嚴重程度的增加，實驗組的組織中逐漸出現(xiàn)中性粒細胞浸潤，成牙本質(zhì)細胞排列紊亂，血管擴張充血等病理性改變；3. RT-PCR法檢測TNF-α結(jié)果表明，與對照組相比，實驗1d組、3d組、5d組的TNF-α mRNA水平的表達量隨著牙髓炎嚴重程度的增加而升高（P0.05）；4.免疫組織化學染色結(jié)果表明：正常組、實驗1d組、3d組、5d組的平均光密度分別為：0.2631±0.0609、0.3628±0.0610、0.5249±0.1815、0.6354±0.2160，實驗組與對照組相比，差異有統(tǒng)計學意義(P＜0.05)；5. Western blot分析結(jié)果表明：正常組、實驗1d組、3d組、5d組的Nav1.3的相對表達強度分別為：0.320±0.041、0.551±0.038、0.983±0.027、1.142±0.019，實驗組與對照組相比，差異有統(tǒng)計學意義(P＜0.05)。 結(jié)論本研究證實大鼠牙髓組織中存在Nav1.3的表達，牙髓炎所致的炎性疼痛可導(dǎo)致Nav1.3表達顯著上調(diào)，這可能是炎性疼痛發(fā)生時神經(jīng)元細胞膜上鈉通道功能異常的分子學基礎(chǔ)之一。鈉通道Nav1.3與牙髓炎痛覺致敏之間可能存在潛在關(guān)聯(lián)，牙髓炎導(dǎo)致炎性疼痛時，Nav1.3表達上調(diào)很可能參與了成牙本質(zhì)細胞的興奮性反應(yīng)，可進一步推測臨床上研制Nav1.3抑制劑，可能有助于臨床上調(diào)整牙痛的治療靶點。
[Abstract]:Objective voltage gated sodium channel subtype Nav1.3 belongs to tetrodotoxin sensitive (TTX-S) sodium channel. Nav1.3 is mainly expressed in rodent embryonic brain tissue. It can quickly activate and quickly deactivate Na+ current. It has a significant regulating action on action potential. It can be re expressed and ectopic after inflammatory stimulation and neuron injury. The sensory work of dental pulp tissue It can be controlled by pain receptors, specific nerve fibers form a pain receptor. Under normal circumstances, appropriate external stimuli do not cause discomfort in dental pulp tissue, but when pulpitis secondary to pulpitis, such as secondary pulpitis, the pain receptor in the pulp tissue will produce pain response. This experiment was made by constructing an experimental rat model of pulpitis. The expression of sodium channel Nav1.3 in pulp tissue of normal rats and mild, moderate, and severe pulpitis rats was compared and analyzed. The relationship between the expression of sodium channel Nav1.3 and inflammatory pain caused by pulpitis was discussed, which could help to adjust the therapeutic target of toothache, and help to find better than the traditional local anesthetic. A new way to relieve toothache.
Methods 16 rats were randomly divided into control group, experimental group 1D, group 3D and group 5D, 4 rats in each group. After Escherichia Coli Endotoxin (LPS) induced rat pulpitis, 1,3,5d was executed after the injury. After the structural model, the animal behavior of rats was observed. HE staining was used to detect the pathological condition of the control group and the experimental 1D, 3D, 5D group of the pulp tissue, and the reversal of the pathological condition of the pulp tissue in the group of 1D, 3D, and 5D. RT-PCR method was used to detect the expression of TNF- alpha mRNA in the pulp tissues of the control group and the experimental group of 1D, 3D and 5D, to determine the relationship between the severity of pulpitis and the inflammatory pain, and to detect the qualitative expression of Nav1.3 in the dental pulp tissues of normal rats and pulpitis rats by immunohistochemical technique and the detection of Western blot method. The relative expression of Nav1.3 protein in dental pulp tissue of normal rats and pulpitis rats was compared.
Results the results of 1. animal behavior observation showed that the diet of normal rats was normal and the mental state was better. The rats in the experimental group appeared repeatedly licking the wound after making the model, did not dare to bite the food, was restless, screamed, and lost weight. The results of 2.HE staining showed that the structure of the normal group was normal, without pathological changes; with the teeth. The severity of myeloid inflammation increased, the tissue of the experimental group gradually appeared neutrophils infiltration, odontoblasts were arranged disorder, vascular dilatation and congestion and other pathological changes, and the results of TNF- alpha detected by 3. RT-PCR method showed that the expression of TNF- alpha mRNA in experimental group 1D, 3D group and 5D group increased with the increase of the severity of pulpitis. The results of 4. immunohistochemical staining showed that the average light density of the normal group, the experimental group 1D, the 3D group and the 5D group were respectively: 0.2631 + 0.0609,0.3628 + 0.0610,0.5249 + 0.2160, and the experimental group was statistically significant (P < 0.05) compared with the control group (P < 0.05), and 5. Western blot analysis showed that the normal group was normal, The relative expression intensity of Nav1.3 in group 1D, group 3D and 5D group was 0.320 + 0.041,0.551 + 0.038,0.983 + 0.019 respectively. The difference was statistically significant (P < 0.05) compared with the control group (P < 0.05).
Conclusion the present study confirmed the expression of Nav1.3 in the pulp tissue of rats, and the inflammatory pain caused by pulpitis can lead to a significant up-regulated expression of Nav1.3. This may be one of the sub molecular basis of the abnormal sodium channel function on the membrane of neuronal cells during the occurrence of inflammatory pain. There may be a potential association between sodium channel Nav1.3 and the sensitization of dental myelomyctis. When pulpitis causes inflammatory pain, the up-regulated expression of Nav1.3 is likely to be involved in the excitatory response of odontoblasts. It is possible to further speculate that the clinical development of Nav1.3 inhibitors may contribute to the therapeutic targets for the clinical adjustment of toothache.

【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R781.31

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