外周和脊髓NMDA受体在Formalin引起的脊髓小胶质细胞活化和长时程痛敏中的作用

发布时间：2018-01-01 23:21

本文关键词：外周和脊髓NMDA受体在Formalin引起的脊髓小胶质细胞活化和长时程痛敏中的作用　出处：《山西医科大学》2005年硕士论文　论文类型：学位论文

【摘要】：经典的formalin 痛模型即脚掌注射formalin 引起的持续约1 小时的双相性痛行为反应。由于能较好地模拟临床急性炎症痛及能采用定性和半定量的指标对痛反应进行测量,因而被广泛应用于疼痛机制的研究。新近的研究显示,formalin 皮下注射除引起上述急性炎性痛外,还可以引起持续性的痛敏状态,如持续约4 周的热辐射痛敏反应及同时观察到的脊髓背角小胶质细胞的激活。在以往的实验中,本研究室及其他研究者的观察都证实,NMDA 受体在formalin 引起的急性炎性痛中发挥重要作用。本实验在外周注射formalin 的基础上,通过痛行为观察(热辐射刺激缩足反射潜伏期)、对小胶质细胞活化增殖的免疫组化检测(OX-42 阳性表达),结合外周及鞘内注射NMDA 受体拮抗剂MK-801,试图证实外周与脊髓NMDA 受体在formalin 所致的长时程痛敏及脊髓小胶质细胞的活化中发挥重要作用。实验在Sprague-Dawley 雄性大鼠(90~100g)进行。本实验分两部分,第一部分是通过外周注入MK-801 观察外周传入感觉神经末稍上NMDA 受体在formalin 引起的长时程痛敏反应及脊髓小胶质细胞活化中的作用。包括免疫组化实验和行为学观察。在免疫组化实验中,根据给药方法的不同,分为多次给药组和单次给药组(n=6)。多次给药组中,在动物两侧脚掌注入formalin 前15 分钟,将NMDA 受体的拮抗剂MK-801(分为三个剂量组即10 nmol、30 nmol、50 nmol)注入左侧脚掌皮下,而在右侧注入生理盐水(NS),并且在formalin 注射后的第1 天、第2 天、第3 天、第4 天分别重复给予原剂量的MK-801或NS。在注射formalin后第7天进行OX-42表达的免疫组化检测,对比背角两侧小胶质细胞的免疫组化反应,进行图像分析及统计学处理。单次给药组也分为三个剂量组即50 nmol、100 nmol、150 nmol,与多次给药组不同的是注射formalin 前15 分钟一次全量给予MK-801 或NS,到第7 天进行免疫组化检测。在痛行为实验观察中,分为实验组(30 nmol MK-801+formalin)和对照组(NS+formalin),采用多
[Abstract]:Diphasic pain behavior response formalin pain model is the classic peripheral injection of formalin induced lasted about 1 hours. Because it can mimic the clinical acute inflammatory pain and can use qualitative and semi quantitative indicators of pain response were measured, so it is widely used in the research of the mechanism of pain. Recent studies have shown that formalin in addition to the subcutaneous injection caused by acute inflammatory pain, can also cause pain sensitive state persistence, activate the heat radiation of pain sensitivity as lasts for 4 weeks and also observed in spinal microglia. In previous experiments, observe the research lab and other researchers have confirmed and play an important role in formalin induced NMDA receptor in acute inflammatory pain. In this experiment the peripheral injection of formalin on pain behavior through the observation (thermal withdrawal latency), the activation of microglia increased Immunohistochemistry (OX-42 positive expression), combined with peripheral and intrathecal injection of NMDA receptor antagonist MK-801, attempts to confirm the role of NMDA receptors in peripheral and spinal cord in the long term pain sensitivity induced by formalin and the activation of spinal microglia.
Experiment in male Sprague-Dawley rats (90~100g). The experiment is divided into two parts. The first part is through the peripheral injection of MK-801 was observed in the peripheral role of Cheng Tongmin reaction and spinal microglia activation of afferent sensory nerve endings of NMDA receptors in formalin induced long. Including immunohistochemical experiments and behavioral observation in the immunohistochemical experiments, according to the different methods, divided into multiple dosing group and single drug group (n=6). Multiple dosing group, formalin injection in animal feet on both sides of the first 15 minutes, the NMDA receptor antagonist MK-801 (divided into three groups: 10 nmol 30, nmol, 50 nmol) subcutaneously injected into the left foot, while the injection of normal saline (NS) on the right side, and on the first day after formalin injection for second days, third days, fourth days respectively to the original repeated doses of MK-801 or NS. in seventh days after the injection of formalin OX-42 The group of immune detection, immune group comparison of dorsal horn microglia on both sides of the reaction, image analysis and statistical treatment. The single dose group was also divided into three groups: 50 nmol, 100 nmol, 150 nmol, and multiple dose group is 15 minutes before the first injection of formalin the full amount of MK-801 or NS, to seventh days by immunohistochemical detection. In pain behavior experiment, divided into experimental group (30 nmol MK-801+formalin) and control group (NS+formalin), the

【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R338

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