电针对脊髓损伤小鼠运动功能及炎性和氧化应激反应的影响  

Electroacupuncture improves locomotor function by regulating expression of inflammation and oxidative stress-related proteins in mice with spinal cord injury

作　　者：代妮 黄思琴[1] 唐成林[1] 谭程方 代攀 曾婷婷 朱正威[1] 杨之雪 DAI Ni;HUANG Si-qin;TANG Cheng-lin;TAN Cheng-fang;DAI Pan;ZENG Ting-ting;ZHU Zheng-wei;YANG Zhi-xue(College of Traditional Chinese Medicine of Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学中医药学院

出　　处：《针刺研究》2019年第11期781-786,804,共7页Acupuncture Research

基　　金：国家自然科学基金青年基金项目(No.81403466);重庆市教委项目(KJ1702033);重庆市科委项目(cstc2017jcyjAX0363、2018jcyjAX0036);重庆市卫计委项目(ZY201802026);人社局留创计划创新类项目(cx2018106)

摘　　要：目的:观察电针对脊髓损伤(SCI)小鼠运动功能的影响,从抗炎及抗氧化方面探讨电针促进SCI小鼠功能修复的机制。方法:雌性C57BL/6小鼠随机分为假手术组、模型组及电针组,每组12只。采用钳夹法制备小鼠SCI模型。电针组于造模后3 h电针双侧"足三里""三阴交",每日1次,连续治疗7 d。采用Basso Mouse Scale(BMS)评分评估小鼠后肢运动功能变化;HE染色法观察脊髓损伤区病理形态学变化;Western blot法检测小鼠脊髓中载脂蛋白E(ApoE)及磷酸化核转录因子-κB(p-NF-κB)、白细胞介素-1β(IL-1β)、细胞外调节蛋白激酶(ERK1/2)、p-ERK1/2、核因子E2相关因子2(Nrf2)、血红素氧化酶-1(HO-1)的表达;免疫荧光染色法检测脊髓中星形胶质细胞的增生程度。结果:造模后第7天,与假手术组比较,模型组小鼠的BMS评分显著下降(P<0. 05);脊髓组织结构紊乱,炎性浸润严重,正常神经元大量减少;脊髓中ApoE、p-NF-κB、IL-1β、p-ERK1/2、Nrf2、HO-1的表达显著升高(P<0. 05);星形胶质细胞数量显著增加(P<0. 05)。与模型组比较,电针组小鼠的BMS评分显著升高(P<0. 05);脊髓组织结构较完整,炎性浸润较轻,正常神经元较多;脊髓中p-NF-κB、IL-1β的表达显著降低(P<0. 05),ApoE、p-ERK1/2、Nrf2、HO-1显著升高(P<0. 05);星形胶质细胞数量显著减少(P<0. 05)。结论:电针能够显著改善SCI小鼠的炎性反应和氧化应激反应,抑制星形胶质细胞过度增生,从而促进脊髓功能的修复,其作用机制可能与上调脊髓中ApoE的表达,增强Nrf2/HO-1抗氧化途径和抑制NF-κB激活有关。Objective To observe the effect of electroacupuncture(EA)on the expression of apolipoprotein E(ApoE)and related proteins of inflammation and anti-oxidative stress in spinal cord in mice with spinal cord injury(SCI),so as to explore its mechanisms underlying function repair.MethodsThirty-six female C57BL/6 mice were equally randomized into 3 groups:sham operation,model and EA. The SCI model was established by clamping the spinal cord for 25s with a serrefine after laminectomy of the 1^st lumbar vertebra(L1). EA(1. 5 Hz/7. 5 Hz,1. 0 mA)was applied to bilateral"Zusanli"(ST36)and"Sanyinjiao"(SP6)for 10 min,once a day for 7 days. The hindlimb locomotor function was assessed according to the state of the range of motion,coordination,claw gesture of the hind leg ankle-joint,trunk stability and the tail posture by using Basso Mouse Scale(BMS). The histopathological changes of the injured area of the spinal cord were determined by H. E. staining. The expression levels of Apo E,phosphorylated nuclear transcription factor-κB(p-NF-κB),interleukin 1 beta(IL-1β),phosphorylated extracellular regulatory protein kinase(p-ERK1/2),extracellular regulatory protein kinase(ERK1/2),nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxidase-1(HO-1)in the spinal cord were detected by Western blot,and the glial fibrillary acidic protein(GFAP)-positive astrocytes were displayed by immunofluorescence staining.ResultsAfter modeling,the BMS scores were significantly decreased in the model group compared with the sham operation group(P<0. 05). Following EA,the BMS scores were markedly increased in the EA group relevant to the model group(P<0. 05),suggesting an improvement of the hindlimb locomotor function. H. E. stain showed structural disorder with lots of cavities,severe inflammatory infiltration with large quantity of inflammatory cells,and apparent reduction of normal neurons in the injured spinal cord tissue of model group,which was milder in the EA group. The expression levels of Apo E,p-NF-κB,IL-1β,p-ERK1/2(not ERK1/2),Nrf2

关 键 词：脊髓损伤 电针 后肢运动功能 载脂蛋白E 核因子E2相关因子2/血红素氧化酶-1抗氧化途径 核转录因子-ΚB 星形胶质细胞 

分 类 号：R245.9+7[医药卫生—针灸推拿学]