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目的:探讨钙/钙调素依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)在神经病理性疼痛中的作用。方法:坐骨神经选择性损伤(SNI)大鼠模型,随机分为4组(n=8):SNI组,Sham组,NS组,AIP组,后两组分别于术前20 min鞘内注射10μl的生理盐水或10%的AIP。于术后1d、2d、3d、4d、6d、8d、10d、14d测定大鼠机械缩足反射阈值(mechanical with-drawal threshold,MWT)。分别于术后1d、3d、7d取大鼠脊髓腰段,用免疫组织化学法(n=6)检测脊髓背角磷酸化pCaMKⅡ的表达;Western blot法(n=4)检测脊髓背角神经元细胞核内pCREB的表达。结果:术后1~3d AIP组大鼠MWT较NS组明显升高(P<0.01);术前鞘内注射AIP在术后1d、3d能够使脊髓背角pCaMKⅡ的表达减少,同时脊髓背角神经元细胞核内pCREB的表达也明显减少。结论:CaMKⅡ参与了神经病理性痛的形成,其作用部分通过CREB介导。
Objective: To investigate the role of calcium / calmodulin-dependent protein kinase Ⅱ (CaMKⅡ) in neuropathic pain. Methods: Rats with sciatic nerve selective injury (SNI) were randomly divided into 4 groups (n = 8): SNI group, sham group, NS group and AIP group. The latter two groups were given intrathecal injection of 10μl Saline or 10% AIP. Rat mechanical at-drawal threshold (MWT) was measured at 1 day, 2 days, 3 days, 4 days, 6 days, 8 days, 10 days and 14 days after operation. Spinal cord lumbar segments were obtained at 1 day, 3 days and 7 days after operation, respectively. The expression of phosphorylated pCaMKII in spinal dorsal horn was detected by immunohistochemical method (n = 6). The expression of pCaMKII in spinal dorsal horn was detected by Western blot (n = 4) Expression of pCREB in the nucleus. Results: The MWT of rats in AIP group was significantly higher than that of NS group (P <0.01) at 1 ~ 3d after operation. The preoperative intrathecal injection of AIP reduced the expression of pCaMKⅡ in spinal dorsal horn 1 day and 3 days after operation, The expression of pCREB in the nucleus of neurons was also significantly reduced. CONCLUSIONS: CaMKII is involved in the formation of neuropathic pain and its role is partly mediated by CREB.