苯肾上腺素预处理后放射损伤早期大鼠颌下腺p-JNK的表达

发布时间：2018-03-16 05:15

本文选题：颌下腺　切入点：放射损伤　出处：《大连医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：对于口腔颌面头颈部恶性肿瘤的患者，不管是术前还是术后放射治疗都已经成为综合序列治疗的重要方法之一。口腔颌面部的两对大唾液腺，腮腺位于面侧深区，颌下腺位于颌下三角内，均与颅颈部解剖位置相接近且易于受到放射线的损害，再加上放射技术的限制，这些特点使得，头颈部恶性肿瘤患者在接受放射治疗时都会损伤到正常的唾液腺组织，只能尽可能地减小对涎腺组织的损伤。虽然患者的5年生存率明显提高，但是患者的生存质量受到严重影响，而且腺体自身修复功能低下，目前尚无有效的治疗方法。因此，作为这一领域的新一代学者，我们应该继续学习和研究放射治疗导致涎腺功能损伤的具体机制，并致力于研究和探索有效的预防和治疗方法。据文献报道，苯肾上腺素（phenylephrine,PE）可明显改善X线照射后早期阶段和晚期阶段涎腺功能的修复，然而其机制尚不明了。据国内外学者报道在JNK信号通路中p-JNK发挥着促进细胞凋亡的作用，然而这一作用在放射性损伤的涎腺组织中是否存在尚未见报道，我们课题组一直研究的苯肾上腺素对其在信号通路中的表达是否会产生影响等诸多问题均为解决。目的探讨苯肾上腺素预处理对放射损伤早期大鼠颌下腺p-JNK蛋白表达的影响。材料和方法将27只Wistar大鼠,随机分为三组：对照组，单纯照射组和苯肾上腺素预处理组。麻醉后对预处理组将苯肾上腺素按照每千克体重5mg的剂量，照射前半小时通过腹腔注射到大鼠体内。将单纯照射和预处理组大鼠分别固定于泡沫板上，使颌下区充分暴露，调整直线加速器的照射野对准大鼠颌下区，给予X线照射剂量为20Gy。在照射后的第7天摘取各组大鼠颌下腺腺体组织，而后应用免疫组织化学方法检测各组大鼠颌下腺组织中p-JNK的表达和分布；应用免疫印迹技术（Western Blot）检测各组大鼠颌下腺p-JNK蛋白及JNK总蛋白的表达。结果利用显微镜观察免疫组织切片所得的结果：空白对照组显示颌下腺组织细胞的胞浆和胞核中仅有少量p-JNK蛋白的表达，单纯照射组磷酸化JNK蛋白在颌下腺细胞的胞质和胞核中的表达明显增加，而苯肾上腺素预处理组中磷酸化JNK蛋白的表达减少，相较于单纯照射组大鼠其在颌下腺细胞胞核及胞质中的表达减少更明显。Western blot的实验结果显示：三组大鼠颌下腺组织中JNK总蛋白的表达尚未见明显差异；单纯照射实验组中p-JNK蛋白的表达显著增加，利用统计学原理与空白对照组的表达作比较，结果可得增加了约40.14%（P＜0.05），按照同样的方法比较，苯肾上腺素预处理组比单纯照射组下降了约45.02%（P＜0.05）。结论放射线照射导致大鼠颌下腺损伤的早期阶段，，可使腺体组织细胞中p-JNK的表达显著增加，然而利用苯肾上腺素进行放射治疗前预防处理，能够下调放射线照射损伤后早期大鼠颌下腺中p-JNK蛋白的表达,又因为苯肾上腺素作用的受体为а1-肾上腺素受体，所以推测活化的а1-肾上腺素受体可能参与了颌下腺放射损伤后的功能调节过程，这为学者们研究行之有效的预防和治疗颌下腺放射损伤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.

【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.8

【参考文献】