本文選題：頜下腺　切入點：放射損傷　出處：《大連醫(yī)科大學》2014年碩士論文　論文類型：學位論文

【摘要】：對于口腔頜面頭頸部惡性腫瘤的患者，不管是術前還是術后放射治療都已經成為綜合序列治療的重要方法之一�？谇活M面部的兩對大唾液腺，腮腺位于面?zhèn)壬顓^(qū)，頜下腺位于頜下三角內，均與顱頸部解剖位置相接近且易于受到放射線的損害，再加上放射技術的限制，這些特點使得，頭頸部惡性腫瘤患者在接受放射治療時都會損傷到正常的唾液腺組織，只能盡可能地減小對涎腺組織的損傷。雖然患者的5年生存率明顯提高，但是患者的生存質量受到嚴重影響，而且腺體自身修復功能低下，目前尚無有效的治療方法。因此，作為這一領域的新一代學者，我們應該繼續(xù)學習和研究放射治療導致涎腺功能損傷的具體機制，并致力于研究和探索有效的預防和治療方法。據文獻報道，苯腎上腺素（phenylephrine,PE）可明顯改善X線照射后早期階段和晚期階段涎腺功能的修復，然而其機制尚不明了。據國內外學者報道在JNK信號通路中p-JNK發(fā)揮著促進細胞凋亡的作用，然而這一作用在放射性損傷的涎腺組織中是否存在尚未見報道，我們課題組一直研究的苯腎上腺素對其在信號通路中的表達是否會產生影響等諸多問題均為解決。 目的探討苯腎上腺素預處理對放射損傷早期大鼠頜下腺p-JNK蛋白表達的影響。 材料和方法將27只Wistar大鼠,隨機分為三組：對照組，單純照射組和苯腎上腺素預處理組。麻醉后對預處理組將苯腎上腺素按照每千克體重5mg的劑量，照射前半小時通過腹腔注射到大鼠體內。將單純照射和預處理組大鼠分別固定于泡沫板上，使頜下區(qū)充分暴露，調整直線加速器的照射野對準大鼠頜下區(qū)，給予X線照射劑量為20Gy。在照射后的第7天摘取各組大鼠頜下腺腺體組織，而后應用免疫組織化學方法檢測各組大鼠頜下腺組織中p-JNK的表達和分布；應用免疫印跡技術（Western Blot）檢測各組大鼠頜下腺p-JNK蛋白及JNK總蛋白的表達。 結果利用顯微鏡觀察免疫組織切片所得的結果：空白對照組顯示頜下腺組織細胞的胞漿和胞核中僅有少量p-JNK蛋白的表達，單純照射組磷酸化JNK蛋白在頜下腺細胞的胞質和胞核中的表達明顯增加，而苯腎上腺素預處理組中磷酸化JNK蛋白的表達減少，相較于單純照射組大鼠其在頜下腺細胞胞核及胞質中的表達減少更明顯。Western blot的實驗結果顯示：三組大鼠頜下腺組織中JNK總蛋白的表達尚未見明顯差異；單純照射實驗組中p-JNK蛋白的表達顯著增加，利用統(tǒng)計學原理與空白對照組的表達作比較，結果可得增加了約40.14%（P＜0.05），按照同樣的方法比較，苯腎上腺素預處理組比單純照射組下降了約45.02%（P＜0.05）。 結論放射線照射導致大鼠頜下腺損傷的早期階段，，可使腺體組織細胞中p-JNK的表達顯著增加，然而利用苯腎上腺素進行放射治療前預防處理，能夠下調放射線照射損傷后早期大鼠頜下腺中p-JNK蛋白的表達,又因為苯腎上腺素作用的受體為а1-腎上腺素受體，所以推測活化的а1-腎上腺素受體可能參與了頜下腺放射損傷后的功能調節(jié)過程，這為學者們研究行之有效的預防和治療頜下腺放射損傷的方法提供了新思路。
[Abstract]:For oral and maxillofacial head and neck cancer patients, whether preoperative or postoperative radiotherapy has become one of the important methods in comprehensive therapy. Two of the major salivary glands in oral and maxillofacial region, parotid gland is located in the deep region of lateral face, the submandibular gland in the submandibular triangle, with craniocervical anatomic location and easy to close radiation damage, plus radiation technology, because of these characteristics, patients with malignant tumor of head and neck injury to normal salivary gland tissue in radiotherapy, only to reduce as much as possible on salivary gland tissue injury. Although the 5 year survival of patients was obviously improved, but the quality of life of patients the severely affected, and the gland self repair function is low, there is no effective treatment. Therefore, as the field of a new generation of scholars, we should continue to study and research of radiation therapy The specific mechanism of salivary gland function damage, and is committed to research and explore effective methods of prevention and treatment. It is reported that phenylephrine (phenylephrine, PE) can repair early stage and late stage of salivary gland function improved significantly after irradiation, but the mechanism is still unclear. According to the domestic and foreign scholars have reported in JNK signal pathway p-JNK plays the role of promoting cell apoptosis, but the role in salivary gland tissue radiation damage has not been reported in the presence of phenylephrine, our research group has been studying on its expression in the signal path will affect many problems are solved.
Objective to investigate the effect of phenylephrine preconditioning on the expression of p-JNK protein in the submandibular gland of rats in early stage of radiation injury.
Materials and methods 27 Wistar rats were randomly divided into three groups: control group, irradiation group and phenylephrine pretreatment group. After anesthesia of phenylephrine pretreatment group will according to the dose of 5mg per kilogram of body weight, half hour before irradiation by intraperitoneal injection into rats. The single pure and pre irradiation the rats of treatment group were fixed on the foam board, the submandibular region fully exposed, irradiation linac alignment adjustment of rat submandibular area, given the X-ray radiation dose from submandibular glands tissue of rats was 20Gy. in seventh days after irradiation, the expression and distribution of p-JNK and application of immunohistochemical method to detect the rat submandibular gland tissues; application of immunoblotting (Western Blot) was used to detect the expression of rat submandibular gland p-JNK protein and JNK protein.
The observation of immune tissue with microscope results: the blank control group showed that the expression of submandibular gland cell cytoplasm and nuclei in only a small amount of p-JNK protein, the expression of the irradiated group of phosphorylated JNK protein in submandibular gland cell nucleus and cytoplasm was significantly increased, while the expression of phenylephrine in pretreatment group the reduction of JNK protein, compared with irradiation group rats and its expression in submandibular gland cell nuclei and cytoplasm decreased significantly.Western blot results showed that the expression of JNK in three groups of rat submandibular gland tissues have not obvious difference; the expression of p-JNK protein in the experimental group was irradiated by simple increased significantly. The expression of the principle of statistics and the control group for comparison, results can be increased by about 40.14% (P < 0.05), compared with the same method, phenylephrine pretreatment group than in pure light The ejection group decreased by about 45.02% (P < 0.05).
Conclusion irradiation leads to early stage of submandibular gland injury in rats, the expression of p-JNK in glandular tissue cells increased significantly, however the use of phenylephrine before radiotherapy for prevention and treatment of radiation injury, can decrease the early p-JNK rat submandibular gland in protein expression, and because the effects of epinephrine receptor for benzene 1- adrenergic receptor that activation of 1- adrenergic receptor may be involved in the regulation process of submandibular gland function after radiation injury, it provides a new idea for effective research the prevention and treatment of radiation damage to the submandibular gland method.

【學位授予單位】：大連醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R739.8

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