nNOS磷酸化在大鼠神经病理性疼痛发病中的作用及其机制  

The mechanism of n NOS phosphorylation in spinal cord of neuropathic pain in rat

作　　者：何庆标[1] 闵宇懿[2] 张绍杰[1] 莫力[1] 王育明[1] 

机构地区：[1]武警广东省总队医院麻醉科,广东广州510507 [2]南方医科大学南方医院儿科,广东广州510515

出　　处：《南京医科大学学报（自然科学版）》2016年第3期302-306,共5页Journal of Nanjing Medical University(Natural Sciences)

摘　　要：目的：探索神经结扎（spinal nerve ligation,SNL）模型大鼠脊髓内神经元型一氧化氮合酶（neuronal nitric oxide synthase,n NOS）的磷酸化是否对神经病理性疼痛具有调节作用,研究n NOS磷酸化在神经病理性疼痛中的作用机制。方法 ：选用雄性SPF级SD大鼠60只,随机分为4组。假手术组：只暴露脊神经,不结扎;实验组：制作SNL模型,鞘内注射钙依赖性蛋白激酶Ⅱ（phosphorylated calcium/calmodulin dependent kinaseⅡ,p-Ca MKⅡ）抑制剂KN93;阴性对照组：制作SNL模型,鞘内注射DMSO;模型组：制作SNL模型,不给药。于术前1 d、术后1~5 d以及鞘内给药后1~4 h测定机械痛阈值。采用Western blot检测腰段脊髓组织p-Ca MKⅡ、n NOS与p-n NOS的表达水平,利用免疫共沉淀以及免疫荧光实验检测n NOS与其接头蛋白CAPON是否具有相互作用。结果：SNL可导致大鼠机械疼痛阈值降低（P〈0.01）,脊髓组织p-Ca MKⅡ的表达增加（P〈0.05）,p-n NOS的表达下降（P〈0.05）,髓鞘内注射KN93可反转神经结扎所致的上述趋势;n NOS与其接头蛋白CAPON在大鼠体内存在相互作用,n NOS的磷酸化能降低其与CAPON的相互作用强度。结论：n NOS的磷酸化参与了SNL诱导的神经病理性疼痛的维持,p-Ca MKⅡ通过对n NOS的磷酸化,降低n NOS与其接头蛋白CAPON在体内的相互作用强度,针对n NOS的磷酸化信号途径的治疗可为神经病理性疼痛的治疗提供新的视野。Objective：To explore the effect of phosphorylation of neuronal nitric oxide synthase（n NOS） on regulation the neuropathic pain in spinal nerve ligation（SNL） model rats,and research the mechanism of n NOS phosphorylation in neuropathic pain. Methods：A total of 60 male SPF SD rats were randomly divided into 4 groups：the control group,only expose spinal nerve,not ligation;the experimental group,SNL model,KN93（calcium / calmodulin dependent kinase Ⅱ inhibitor） intrathecal injection;the negative control group,SNL model,DMSO intrathecal injection;the model group,SNL model,didn′t receive any medicine. We determined mechanical pain threshold 1 day before operation,postoperative 1-5 days and 1-4 hours after sheath dosing. Western blot was performed to detect the expression level of calcium / calmodulin dependent kinase Ⅱ（p-Ca MKⅡ）,n NOS and pn NOS in lumbar spinal cord tissue. Co IP and immunofluorescence experiments was performed to test whether there was an interaction between n NOS and CAPON. Results：SNL led to mechanical pain threshold of rats reduced（P〈0.01）,the expression of spinal cord tissue p-Ca MK Ⅱ increased（P〈0.05）,and the expression of p-n NOS decreased（P〈0.05）. KN93 intrathecal injection reversed the trend above. There was an interaction between n NOS and CAPON,and the phosphorylation of n NOS reduced the strength of interaction between n NOS and CAPON. Conclusion：The phosphorylation of n NOS is involved in neuropathic pain induced by SNL,and p-Ca MK Ⅱ make n NOS phosphorylation,reduced the strength of interaction between n NOS and CAPON. The n NOS signaling pathways can provide a new field of vision for neuropathy pain treatment.

关 键 词：NNOS CAPON 磷酸化 神经病理性疼痛 

分 类 号：R614.1[医药卫生—麻醉学]