水通道蛋白4基因敲除对神经病理性疼痛的影响及可能机制研究  

Effect of aquaporin 4 gene knockout on neuropathic pain and its possible mechanism

作　　者：卢关伊 庞冲 陈文强 陈瑛 吴宁 李锦 LU Guan-yi;PANG Chong;CHEN Wen-qiang;CHEN Ying;WU Ning;Li Jin(Beijing Key Laboratory of Neuropsychopharmacology,State Key Laboratory of Toxicology and Medical Countermeasures,Institute of Pharmacology and Toxicology,Academy of Military Medicine,Academy of Military Sciences,Beijing 100850,China)

机构地区：[1]军事科学院军事医学研究院毒物药物研究所,抗毒药物与毒理学国家重点实验室,神经精神药理学北京市重点实验室,北京100850

出　　处：《国际药学研究杂志》2019年第12期918-923,共6页Journal of International Pharmaceutical Research

基　　金：国家自然科学基金资助项目(81171046)。

摘　　要：目的研究水通道蛋白4(AQP4)在神经病理性疼痛中的作用,并探讨其与脊髓星形胶质细胞激活和促炎细胞因子释放的关系。方法应用AQP4基因敲除(KO)和野生型(WT)小鼠建立坐骨神经分支损伤模型(SNI),观察AQP4基因敲除对疼痛行为的影响,随后采用Western印迹法(WB)检测SNI模型小鼠脊髓中星形胶质细胞激活相关标志物胶质原纤维酸性蛋白(GFAP)表达的变化,采用ELISA法检测SNI模型小鼠脊髓中促炎细胞因子(TNF-α、IL-6)含量的变化。结果 SNI手术后,与WT组相比,KO组小鼠的机械刺激痛觉超敏程度显著降低(P<0.01),而经假手术(Sham)处理的WT与KO组小鼠无差异(P>0.05),提示AQP4基因敲除可减轻神经病理性疼痛。WB结果显示,后术14 d,与WT-Sham组相比,WT-SNI组小鼠脊髓GFAP含量显著高升高(P<0.01);与WTSNI组相比,KO-SNI组小鼠脊髓GFAP含量显著降低(P<0.01),提示AQP4基因敲除抑制了小鼠SNI模型脊髓星形胶质细胞的激活。ELISA结果显示,术后14 d,与WT-Sham组相比,WT-SNI组小鼠脊髓促炎细胞因子TNF-α和IL-6含量均显著升高(P<0.05,P<0.01);与WT-SNI组相比,KO-SNI组小鼠脊髓TNF-α和IL-6的含量均显著降低(P<0.05,P<0.01),提示AQP4基因敲除可降低小鼠SNI模型脊髓促炎细胞因子水平。结论 AQP4基因敲除可能通过降低脊髓中星形胶质细胞的激活和促炎细胞因子的生成而减轻神经病理性疼痛。Objective To investigate the effect of aquaporin 4(AQP4)in neuropathic pain and explore the relationship with the activation of spinal astrocytes and release of proinflammatory cytokines.Methods The effect of AQP4 gene knockout(KO)on the pain-related behavior was investigated using the sciatic nerve branch injury model(SNI)of AQP4 KO and wild type(WT)mice.The expression of astrocyte activation-related protein,glial fibrillary acidic protein(GFAP),and the level of proinflammatory cytokines,TNF-αand IL-6,all in the mouse spinal cord samples,were detected by Western blot(WB)and ELISA,respectively.Results After SNI surgery,compared with the WT group,a significant attenuation in mechanical allodynia was found in the KO group(P<0.01),however,no difference was detected between two sham groups(Sham)of WT and KO mice(P>0.05).These results indicated that the AQP4 gene knockout relieved neuropathic pain.Fouteen days after SNI surgery,WB results showed that the GFAP level in mouse spinal cord was significantly higher in the WT-SNI group than in the WT-Sham group(P<0.01),whereas the GFAP level in the KOSNI group was significantly lower than that in the WT-SNI group(P<0.01).These results indicated that AQP4 KO inhibited activation of spinal astrocytes in SNI model mice.In addition,14 days after SNI surgery,ELISA results showed that the levels of mouse spinal cord proinflammatory cytokines,TNF-αand IL-6 in the WT-SNI group were significantly higher than those in the WT-Sham group(P<0.05 and P<0.01,respectively),whereas the levels of TNF-αand IL-6 in the KO-SNI group were significantly lower than those in the WT-SNI group(P<0.05 and P<0.01,respectively).These results indicated that AQP4 KO inhibited the level of mouse spinal cord proinflammatory cytokines in SNI model mice.Conclusion The AQP4 gene knockout may relieve the neuropathic pain via the inhibition of the astrocyte activation and proinflammatory cytokine release.

关 键 词：水通道蛋白4 神经病理性疼痛 星形胶质细胞 胶质原纤维酸性蛋白 促炎细胞因子 

分 类 号：R964[医药卫生—药理学]