PKC参与大鼠慢性偏头痛的中枢敏化机制

发布时间：2018-06-09 00:44

本文选题：蛋白激酶C + 慢性偏头痛　；参考：《重庆医科大学》2017年硕士论文

【摘要】：背景:偏头痛是一种常见的原发性头痛,具有反复发作的特点,头痛具有单侧性、搏动性、程度中到重度、日常活动加重等特点,伴随有恶心、呕吐、畏光和畏声等自主神经症状。偏头痛发病率较高,其反复发作严重影响患者的工作和生活质量,给个人、家庭和社会带来了严重的负担,已被WHO列为四大最严重的慢性功能障碍性疾病之一。偏头痛根据其发作频率可分为发作性偏头痛(episodic migraine,EM)和慢性偏头痛(chronic migraine,CM)。慢性偏头痛的临床诊断标准为:每月头痛发作大于15天,持续三个月且无药物滥用。慢性偏头痛是偏头痛常见的并发症,每年大约有2.5%的发作性偏头痛转化为慢性偏头痛。CM较EM给患者个人、家庭及社会带来了更为严重的负担。所以如何防止偏头痛的慢性化是神经病学领域的一重大难点。近年来,有许多研究发现在偏头痛的慢性化中,中枢敏化扮演者重要角色,故进一步研究偏头痛的发中枢敏化机制,抑制偏头痛的慢性化转归,是寻求偏头痛新的治疗靶点的重要方法,对于提高偏头痛患者的生存质量有着十分重要的意义。既往的研究发现,在多种疼痛模型中,PKC活性显著增高,而抑制PKC的活性后往往可以明显减轻疼痛,提示PKC在疼痛的中枢敏化形成过程中发挥着重要作用。但是PKC是否参与慢性偏头痛的中枢敏化机制,尚无相关研究报道,故本文旨在研究PKC是否通过中枢敏化参与慢性偏头痛病理生理过程。方法:1、实验分组:SD雄性大鼠随机分为7组。A、假手术组(n=11),硬膜外连续7天滴注PBS2μl。B、模型组(n=12),硬膜外连续7天滴注IS2μl。C、模型+溶剂组(n=8),硬膜外连续7天滴注IS2μl,侧脑室注射DMSO5μl。D、PKC抑制剂低剂量组(n=11),硬膜外连续7天滴注IS2μl,侧脑室注射CHE4μg(5μl)。E、PKC抑制剂高剂量组(n=10),硬膜外连续7天滴注IS2μl,侧脑室注射CHE8μg(5μl)。F、PKC激动剂低剂量组(n=10),硬膜外连续7天滴注IS2μl,侧脑室注射PMA100ng(5μl)。G、PKC激动剂高剂量组(n=11),硬膜外连续7天滴注IS2μl,侧脑室注射PMA200ng(5μl)。2、硬膜外置管,手术后两天,选择伤口未感染,导管未堵塞的大鼠进入下一步实验。给予IS或PBS缓慢硬脑膜外滴注,重复7天。模拟硬脑膜痛觉感受器的反复激活,建立慢性偏头痛大鼠模型。3、慢性偏头痛大鼠模型建立后,侧脑室给予PKC抑制剂CHE和PKC激动剂PMA,使用Von Frey test测定大鼠面部及后足的机械痛阈值,Western blot和免疫荧光检测TNC部位PKC、CGRP、c-Fos的蛋白的定位和表达变化。结果:1、“炎性汤”反复刺激硬脑膜后,大鼠面部及足底的机械痛阈值显著降低,而三叉神经脊束尾核部位的CGRP、c-Fos及PKC蛋白表达显著增加。2、使用CHE抑制PKC可以显著增高大鼠面部及足底痛阈值,显著降低CGRP、c-Fos的表达。3、而使用PMA激活PKC则降低痛阈值,显著增高CGRP、c-Fos的表达。结论:1、利用炎性汤反复刺激硬脑膜模拟大鼠慢性偏头痛的痛觉感受器反复激活模型成功。2、PKC可以调节机械痛阈值以及CGRP、c-Fos的表达变化,提示PKC参与慢性偏头痛中枢敏化的病理生理过程。
[Abstract]:Background: migraine is a common primary headache with recurrent episodes. Headache is characterized by unilateral, pulsating, moderate to severe and aggravation of daily activity. It is accompanied by nausea, vomiting, photophobia and awe. The incidence of migraine is high, and its recurrent attacks seriously affect the work and quality of life of the patients. It has brought a serious burden to individuals, families and society, which has been listed as one of the four most serious chronic dysfunction diseases of the WHO. Migraine can be divided into episodic migraine (EM) and chronic migraine (chronic migraine, CM) according to the frequency of its attack. The clinical diagnostic criteria for chronic migraine are: a large monthly headache attack. Chronic migraine is a common complication of migraine on 15 days. Chronic migraine is a common complication of migraine. About 2.5% of the migraine migraine into chronic migraine every year is more serious than EM for individuals, family and society. So how to prevent the chronicity of migraine is a major problem in the field of Neurology. In recent years, many studies have found that central sensitization plays an important role in the chronicity of migraine. Therefore, further study of the central sensitization mechanism of migraine and the inhibition of the chronicity of migraine is an important way to seek new targets for migraine, and it is very important to improve the quality of life of migraine patients. Previous studies have found that in a variety of pain models, PKC activity is significantly increased, and the inhibition of PKC activity can significantly reduce pain, suggesting that PKC plays an important role in the process of central sensitization of pain. But whether PKC is involved in the central sensitization mechanism of chronic migraine has not been reported, so this article is aimed at this article. Study whether PKC is involved in the pathophysiological process of chronic migraine through central sensitization. Methods: 1, the experimental group: SD male rats were randomly divided into 7 groups of.A, sham operation group (n=11), PBS2 mu l.B, model group (n=12), 7 days of epidural continuous infusion of IS2 u l.C, model + solvent group (n=8), 7 days of epidural continuous infusion of IS2 micron, lateral ventricle injection. DMSO5 mu l.D, PKC inhibitor low dose group (n=11), IS2 Mu L for 7 days in epidural continuous infusion, CHE4 mu g (5 L).E for lateral ventricle, PKC inhibitor in high dose group (n=10). C agonist high dose group (n=11), 7 days of epidural continuous infusion of IS2 Mu L, lateral ventricle injection of PMA200ng (5 L).2, extradural catheter, two days after the operation, select the wound not infected, the catheter unblocked rats enter the next experiment. Give IS or PBS slow dural infusion, repeat 7 days. Simulation of the reactivation of the dura meninges, the establishment of a slow activation, establish slow, slow establishment of the dura. .3, a rat model of migraine, after the establishment of a chronic migraine rat model, the lateral ventricle gave the PKC inhibitor CHE and PKC agonist PMA. Von Frey test was used to determine the mechanical pain threshold of the rat's face and hind feet. Western blot and immunofluorescence test TNC site PKC. After stimulating the dura, the threshold of mechanical pain in the face and foot of the rat decreased significantly, while the expression of CGRP, c-Fos and PKC protein in the nucleus of the trigeminal nucleus was significantly increased by.2. The pain threshold of the face and foot in the rat could be significantly increased with the use of CHE to inhibit PKC, and the CGRP, c-Fos expression was significantly reduced, while PMA activated PKC decreased the pain threshold, and the pain threshold was reduced significantly. Significant reduction of pain threshold was made with PMA activation. Significant pain threshold was reduced significantly. Increase the expression of CGRP and c-Fos. Conclusion: 1,.2 is successfully used to stimulate the model of chronic migraine in the dural mimic rats by repeated stimulation of the inflammatory soup. PKC can regulate the threshold of mechanical pain and the changes in the expression of CGRP and c-Fos, suggesting that PKC is involved in the pathophysiological process of central sensitization of chronic migraine.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R747.2

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