摘要
目的探讨脊髓背根神经节(dorsal root ganglion,DRG)中转录因子锌指X连锁复制C(zinc finger X-linked duplicated C,ZXDC)介导CCL2/CCR2信号通路对慢性压迫性神经损伤(chronic construction injury model,CCI)诱导神经性疼痛的作用及机制。方法构建小鼠坐骨神经慢性压迫性损伤模型,免疫荧光染色、Western blot法和实时荧光定量PCR(RT-qPCR)检测正常情况和CCI造模后DRG中ZXDC及CCL2表达变化;将实验动物分为假手术(Sham)组、CCI+AAV-NC组和CCI+AAV-ZXDC siRNA组;Western blot法和免疫荧光染色检测各组小鼠CCI造模后各时间点DRG中ZXDC、CCL2和CCR2表达,RT-qPCR检测DRG中促炎性细胞因子TNF-α和IL-1βmRNA表达,机械性缩足反射测试检测神经性疼痛行为改变。结果ZXDC表达定位在DRG大、中、小神经元。CCI损伤后1~3天DRG中ZXDC和CCL2蛋白和mRNA表达显著增高,CCI后7天二者表达显著降低,ZXDC和CCL2mRNA表达量呈正相关(P均<0.05)。CCI后3天,与Sham组比较,CCI+AAV-NC组和CCI+AAV-ZXDC siRNA组ZXDC、CCL2、CCR2蛋白表达、TNF-α和IL-1βmRNA表达显著增高,其中CCI+AAV-ZXDC siRNA组较CCI+AAV-NC组ZXDC、CCL2、CCR2蛋白表达、TNF-α和IL-1βmRNA显著降低(P均<0.05)。与Sham组比较,CCI+AAV-ZXDC siRNA组和CCI+AAV-NC组CCI后各时间点机械缩足阈值均显著降低,其中CCI后7天,CCI+AAV-ZXDC siRNA组机械缩足阈值显著高于CCI+AAV-NC组(P均<0.05)。结论脊髓背根神经节ZXDC基因敲减通过抑制CCL2/CCR2信号通路介导CCI诱导的神经性疼痛。
Objective To explore the effect and mechanism of C-C motif chemokine ligand 2(CCL2)/C-C chemokine receptor type 2(CCR2)signaling pathway mediated by ZXDC in spinal dorsal root ganglion(DRG)on neuropathic pain after chronic compressive injury.Methods A chronic compressive injury(CCI)mouse model was established.The expression of ZXDC and CCL2 in DRG was detected by immunofluorescence,Western blot,and real-time fluorescent quantitative PCR(RT-qPCR).The animals were divided into sham group,CCI+AAV-NC group,and CCI+AAV-ZXDC siRNA group.Western blot and immunofluorescence were employed to measure the expression of ZXDC,CCL2,and CCR2 in DRG after CCI,and the expression of pro-inflammatory factor TNF-αand IL-1βmRNA was evaluated by RT-qPCR.At last,the paw withdrawal threshold was used to evaluate the changes in neuropathic pain behavior.Results ZXDC was localized in large,medium,and small DRG neurons.The expression of ZXDC and CCL2 protein and mRNA were significantly increased 1-3days after CCI,and decreased 7days after CCI in DRG.The expression of ZXDC and CCL2mRNA was positively correlated(P<0.05).3days after CCI,compared with sham group,ZXDC,CCL2,CCR2 protein expression,TNF-αand IL-1βmRNA in CCI+AAV-NC group and CCI+AAV-ZXDC siRNA group were significantly increased,and ZXDC,CCL2,CCR2 protein expression,TNF-αand IL-1βmRNA in CCI+AAV-ZXDC siRNA group were significantly decreased than those in CCI+AAV-NC group(P<0.05).Compared with sham group,the paw withdrawal threshold of CCI+AAV-ZXDC siRNA group and CCI+AAV-NC group were significantly decreased at various time points after CCI,and the withdrawal threshold in CCI+AAV-ZXDC siRNA group was significantly increased than that in CCI+AAV-NC group at 7days after CCI(P<0.05).Conclusion Spinal dorsal root ganglion ZXDC siRNA can inhibit neuropathic pain after CCI injury by downregulating CCL2/CCR2signaling axis.
作者
李文媛
王晓宇
吕忠孝
刘东明
李艺
王淑影
王莹
LI Wenyuan;WANG Xiaoyu;LV Zhongxiao(Institute of Neural Tissue Engineering,Mudanjiang Medical University,Heilongjiang 157011,China)
出处
《医学研究杂志》
2024年第1期56-62,共7页
Journal of Medical Research
基金
国家自然科学基金资助项目(81870977)
黑龙江省自然科学基金资助项目(JQ2021H004)
黑龙江省属高校基本科研业务费项目(2021-KYYWF-0469)
牡丹江医学院科学基金火炬计划项目(2022-MYHJ-012)
牡丹江医学院研究生导师科研专项计划项目(YJSZX2022007)。
