基于PKC-P2X3信号通路观察电针对神经病理性疼痛小鼠神经修复的作用机制  

Observation of mechanism of electroacupuncture on nerve repair in mice with neuropathic pain based on PKC-P2X3 signal pathway

作　　者：王立 程仕萍[2] 易惺钱[3] 周平生[4] 刘静[5] 陶添明 陈丽 WANG Li;CHENG Shiping;YI Xingqian;ZHOU Pingsheng;LIU Jing;TAO Tianming;CHEN Li(School of Traditional Chinese Medicine,Nanchang Medical College,Nanchang 330004,China;Dermatology Depart-ment of Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine,Nanchang 330006,China;Pe-diatrics,Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine,Nanchang 330006,China;School of International Education,Jiangxi University of Traditional Chinese Medicine,Nanchang 330004,China;Hospital Office of Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine,Nanchang 330006,China)

机构地区：[1]南昌医学院中医学院,南昌330004 [2]江西中医药大学附属医院皮肤科,南昌330006 [3]江西中医药大学附属医院儿科,南昌330006 [4]江西中医药大学国际教育学院,南昌330004 [5]江西中医药大学附属医院办公室,南昌330006

出　　处：《中国免疫学杂志》2024年第6期1178-1182,1188,共6页Chinese Journal of Immunology

摘　　要：目的:基于蛋白激酶C(PKC)-配体门控型非选择性离子通道3(P2X3)信号通路观察电针对神经病理性疼痛小鼠神经修复的作用机制。方法:48只小鼠随机分成假手术组、模型组、针刺组与电针组,每组12只。除假手术组外,其余小鼠构建坐骨神经慢性限制性损伤(CCI)模型。术后第8天针刺组与电针组接受针刺与电针干预,连续7 d。于术前、术后3 d、5 d、7 d、10 d、12 d、14 d对各组进行机械缩足反射阈值(MWT)与热刺激缩足反射潜伏期(TWL)测试。术后15 d处死所有小鼠,HE染色观察坐骨神经组织形态,ELISA检测脊髓组织IL-1β、TNF-α水平,Western blot检测脊髓组织PKC、P2X3水平,免疫荧光检测脊髓神经元PKC、P2X3表达情况。结果:假手术组神经元细胞大小不一,细胞膜完整,细胞质呈细小颗粒状,细胞核大而圆,核仁居中清晰可见;模型组可见萎缩的神经元,有髓神经纤维紊乱,轴突肿胀,神经元间形成间隙;与模型组相比,针刺组、电针组萎缩神经元数量减少(P<0.05),有髓神经纤维排布情况改善,电针组较针刺组改善更为明显。与相同时间点的假手术组相比,术后3 d、5 d、7 d、10 d、12 d、14 d模型组、针刺组、电针组患侧后肢的MWT、TWL值明显下降(P<0.05),IL-1β、TNF-α、PKC、P2X3表达量、PKC、P2X3阳性细胞数、平均光密度值升高(P<0.05);与模型组相比,术后10 d、12 d、14 d针刺组、电针组MWT、TWL值明显上升(P<0.05),IL-1β、TNF-α、PKC、P2X3表达量、PKC、P2X3阳性细胞数、平均光密度值降低(P<0.05);与针刺组相比,术后10 d、12 d、14 d电针组MWT、TWL值升高(P<0.05),IL-1β、TNF-α、PKC、P2X3表达量、PKC、P2X3阳性细胞数、平均光密度值降低(P<0.05)。结论:电针可能减轻CCI模型小鼠神经炎症反应,降低PKC、P2X3表达水平,改善坐骨神经细胞形态与有髓神经纤维排布情况,缩小神经元间间隙,提升MWT、TWL值,缓解神经病理性疼痛程度。Objective:To observe the mechanism of electroacupuncture on nerve repair in mice with neuropathic pain based on the protein kinase C(PKC)-purinergic receptor P2X ligand-gated ion channel 3(P2X3)signal pathway.Methods:A total of 48 mice were randomly divided into sham group,model group,acupuncture group and electroacupuncture group,with 12 mice in each group.Except the sham group,the other mice were constructed with chronic constriction injury(CCI)model.On the 8th day after operation,acupuncture group and electroacupuncture group received acupuncture and electroacupuncture interventions for 7 days.Mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)were measured before operation and 3rd,5th,7th,10th,12th and 14th day after operation.All mice were killed 15 days after operation.The histomorphology of sciatic nerve was ob-served by HE staining.The levels of IL-1βand TNF-αin spinal cord were detected by ELISA.PKC and P2X3 levels in spinal cord tis-sue were detected by Western blot.PKC and P2X3 expressions in spinal cord neurons were detected by Immunofluorescence.Results:In the sham group,the neuronal cells were of different sizes,the cell membrane was intact,the cytoplasm was fine and granular,the nucleus was large and round,and the nucleolus was clearlyvisible in the middle.Atrophic neurons could be seen in the model group,myelinated nerve fibers were disordered,axons were swollen,and spaces were formed between neurons.Compared with the model group,the number of atrophic neurons decreased in the acupuncture group and the electroacupuncture group(P<0.05),the arrange-ment of myelinated nerve fibers improved and the improvement in the electroacupuncture group was more obvious than that in the acu-puncture group.Compared with the sham group at the same time point,on the 3rd,5th,7th,10th,12th and 14th day after opera-tion,the MWT and TWL values of the model group,acupuncture group and electroacupuncture group decreased significantly(P<0.05),while the expressions of IL-1β,TNF-α,PKC,P2X3,the numbers

关 键 词：蛋白激酶C 配体门控型非选择性离子通道3 电针 神经病理性疼痛 脊髓 

分 类 号：R245.3[医药卫生—针灸推拿学]