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目的通过兴奋及抑制蛋白激酶C(protein kinase C,PKC)的活性观察其对福尔马林炎性痛及痛觉过敏时大鼠脊髓后角一氧化氮合酶(nitric oxide synthase,NOS)尤其是对诱生型NOS(induciable NOS,iNOS)的影响,以探讨在该过程中PKC的作用机制。方法将实验动物72只分9组,每组8只,分别为正常组、福尔马林12h组、福尔马林24h组、福尔马林12h加生理盐水组、福尔马林24h加生理盐水组、福尔马林12h加佛波醇脂(phobol-12-myristate-3-acetate,PMA)组、福尔马林24h加PMA组、福尔马林12h加灯盏花素乙(chelerythrin chloride,CH,sigma)组和福尔马林24h加CH组。分别于注射福尔马林后12、24h将大鼠麻醉后取材,采用免疫组织化学方法观察脊髓后角iNOS阳性神经元数目及染色深度,以探讨PKC对NOS的影响。结果与福尔马林12h组相比,福尔马林12h加PMA组iNOS阳性细胞数明显增加(P<0.01),神经细胞及神经纤维染色也明显加深(P<0.01);福尔马林12h加CH组iNOS阳性细胞数明显减少(P<0.01),神经细胞及神经纤维染色明显变浅(P<0.01)。结论鞘内注射PKC兴奋剂PMA能显著增加L_5脊髓后角神经元NOS的活性;鞘内注射PKC抑制剂CH能显著抑制L_5脊髓后角神经元iNOS的活性。在福尔马林炎性痛及痛觉过敏中,脊髓后角iNOS活性在一定程度上受PKC调控。
OBJECTIVE: To investigate the inhibitory effect of excitatory and inhibitory activities of protein kinase C (PKC) on the expression of nitric oxide synthase (NOS) in rat spinal cord during inflammatory pain and hyperalgesia, Inducible NOS (iNOS) in order to explore the mechanism of action of PKC in this process. Methods 72 experimental animals were divided into 9 groups with 8 rats in each group, which were normal group, formalin 12h group, formalin 24h group, formalin 12h group and formalin group 24h Saline, formalin for 12h, phobol-12-myristate-3-acetate (PMA) group, formalin for 24h plus PMA group, formalin for 12h and chelerythrin chloride, CH, sigma) group and formalin for 24 h plus CH group. The rats were anesthetized after formalin injection for 12 and 24 hours respectively. The number and depth of iNOS positive neurons in the dorsal horn of spinal cord were observed by immunohistochemistry to explore the effect of PKC on NOS. Results Compared with formalin group, the number of iNOS positive cells in formalin-treated group increased significantly (P <0.01) and the number of nerve cells and nerve fiber significantly increased in formalin-treated group (P <0.01) The number of iNOS positive cells in CH group decreased significantly at 12h (P <0.01), and the staining of neurons and nerve fibers became lighter (P <0.01). Conclusion Intrathecal injection of PKC agonist PMA can significantly increase the activity of NOS in L_5 spinal dorsal horn neurons; intrathecal injection of PKC inhibitor CH can significantly inhibit iNOS activity in L_5 spinal dorsal horn neurons. In formalin inflammatory pain and hyperalgesia, spinal cord iNOS activity is to some extent regulated by PKC.