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目的研究脊髓星形胶质细胞D-型氨基酸氧化酶(DAAO)/H2O2介导吗啡镇痛耐受的神经生物学作用。方法在全身或脊髓鞘内连续7 d注射吗啡建立吗啡耐受动物模型的同时长期给予或单次给予DAAO抑制剂或沉默脊髓DAAO基因siRNA后,进行5%福尔马林刺激,考察在小鼠或大鼠福尔马林(第Ⅰ相急性疼痛和第II相中枢敏感化疼痛)、热板和甩尾疼痛模型中吗啡镇痛作用耐受性的变化。结果皮下或脊髓联合给予吗啡和一系列结构不同的DAAO抑制剂如CBIO,AS057278和苯甲酸钠,在小鼠/大鼠急性疼痛(热板、甩尾)和福尔马林疼痛模型(第I相急性疼痛和第II相中枢敏感化疼痛)能完全预防吗啡对镇痛的耐受作用,单次剂量可完全逆转已形成的吗啡耐受状态,且DAAO抑制剂预防和逆转吗啡镇痛耐受作用与脊髓DAAO活性抑制成正性相关;脊髓注射siRNA/DAAO完全预防吗啡耐受;吗啡疼痛耐受形成后,脊髓过氧化氢升高15.1%;同时脊髓给予ROS非特异性清除剂PBN亦可完全阻断福尔马林II相疼痛和吗啡镇痛耐受状态。结论脊髓星形胶质细胞DAAO通过生成过氧化氢介导吗啡镇痛耐受性的形成,是阻断吗啡耐受的一个潜在性新靶点分子,为吗啡镇痛耐受揭示一条新的传导通路。本研究结果还表明DAAO至少是脊髓星形胶质细胞介导吗啡镇痛耐受的分子之一,为开发DAAO抑制剂作为新型镇痛药物以及单独使用或与吗啡合用拮抗吗啡耐受提供了药理学基础。
Objective To investigate the neurobiological effects of morphine analgesia-induced tolerance of D-type amino acid oxidase (DAAO) / H2O2 in spinal cord astrocytes. Methods Morphine-tolerant animal models were established by morphine injection for 7 days on the whole body or intrathecally in the spinal cord to establish long-term or single-dose administration of DAAO inhibitor or silencing of DAAO gene in the spinal cord. After 5% formalin stimulation, Or rat formalin (phase I acute pain and phase II central sensitization pain), morphine analgesia tolerance in hot plate and tail-flick pain models. RESULTS: Morphine and a series of structurally distinct DAAO inhibitors such as CBIO, AS057278 and sodium benzoate were administered subcutaneously or in the spinal cord in mice / rats with acute pain (hot plate, tail flick) and formalin pain models (Phase I Acute pain and central sensitization in Phase II) completely prevented morphine’s tolerance to analgesic pain. A single dose completely reversed the morphine-tolerant state that had been formed, and DAAO inhibitors prevented and reversed morphine analgesic tolerance And spinal cord DAAO activity was positively correlated; spinal cord injection of siRNA / DAAO completely prevent morphine tolerance; morphine pain tolerance formation, spinal cord hydrogen peroxide increased by 15.1%; the spinal cord given ROS non-specific scavenger PBN can be completely blocked Formalin Phase II Pain and Morphine Pain Relief State. Conclusions DAAO, a spinal cord astrocyte, mediates the formation of morphine analgesia tolerance through the formation of hydrogen peroxide and is a potential new target molecule that blocks morphine tolerance, revealing a new pathway for morphine analgesia tolerance path. The results of this study also show that DAAO is at least one of the molecules that mediate morphine analgesia tolerance by spinal astrocytes and provides pharmacological evidence for the development of DAAO inhibitors as new analgesics and antagonism of morphine tolerance either alone or in combination with morphine Fundamentals of learning.