电针对膝关节骨关节炎慢性痛大鼠脊髓背角及海马离子钙接头蛋白1表达的影响  

Effect of electroacupuncture on the expression of Iba-1in spinal dorsal horn and hippocampus of chronic pain rats with knee osteoarthritis

作　　者：郑洁 杨锋[3] 袁普卫[3] 樊静杰 郑晓慧 朱芮 李杨乐 ZHENG Jie;YANG Feng;YUAN Pu-wei;FAN Jing-jie;ZHENG Xiao-hui;ZHU Rui;LI Yang-le(Institution of Acupuncture-moxibustion and Tuina,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province,China;Shaanxi Key Laboratory of Acupuncture and Herbal Medicine,Xianyang 712046,Shaanxi Province;First Clinical Medical College,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province)

机构地区：[1]陕西中医药大学针灸推拿学院,陕西咸阳712046 [2]陕西省针药结合重点实验室,陕西咸阳712046 [3]陕西中医药大学第一临床医学院,陕西咸阳712046

出　　处：《针刺研究》2023年第5期431-437,共7页Acupuncture Research

基　　金：国家自然科学基金项目(No.81804162);陕西省科技厅重点研发计划一般项目(No.2022SF-184);陕西省教育厅青年创新团队科研项目(No.21JP034);陕西高校青年创新团队建设项目(No.2019);陕西中医药大学学科创新团队建设项目(No.2019-YL02)。

摘　　要：目的:观察电针干预对膝关节骨关节炎(KOA)慢性痛大鼠痛行为学、海马形态学、脊髓背角及海马炎性细胞因子含量和离子钙接头蛋白1(Iba-1)表达、海马脑源性神经营养因子(BDNF)表达的影响,探讨电针治疗KOA慢性痛的中枢神经机制。方法:SD雌性大鼠随机分为空白组、盐水组、模型组、电针组,每组10只。采用左膝关节腔注射谷氨酸钠碘乙酸法复制KOA慢性痛大鼠模型。电针组于左侧“阳陵泉”“内膝眼”行电针刺激,15 min/次,1次/d, 5次为1个疗程,共治疗2个疗程。造模前1 d,造模第7、14、20、26天进行痛行为学评价。HE染色法观察左侧海马CA1区形态学变化,ELISA法检测左侧腰(L)3—L5脊髓背角及海马CA1区白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α含量,免疫荧光染色法检测左侧脊髓背角及海马CA1区Iba-1的表达,Western blot法检测左侧海马CA1区BDNF的蛋白表达。结果:与空白组比较,模型组大鼠造模后各时点机械痛缩足阈值(MWT)和后肢负重能力显著下降(P<0.01);海马CA1区神经元排列散乱且数量明显减少,细胞轮廓不清晰,细胞核与胞质界限不清楚,存在核固缩现象;脊髓背角和海马CA1区IL-1β、TNF-α含量显著升高(P<0.01),Iba-1表达显著上调(P<0.01);海马CA1区BDNF的蛋白表达显著下调(P<0.01)。电针干预后,与模型组比较,电针组MWT和后肢负重能力显著升高(P<0.01);海马CA1区神经元在形态、完整性和排列方面均明显优于模型组;脊髓背角及海马CA1区IL-1β、TNF-α含量显著降低(P<0.01),Iba-1表达显著下调(P<0.01);海马CA1区BDNF的蛋白表达显著上调(P<0.01)。结论:电针“阳陵泉”“内膝眼”可缓解KOA大鼠的痛行为表现,其机制可能与电针抑制脊髓背角及海马小胶质细胞介导的炎性反应,上调海马BDNF的表达有关。Objective To observe the effect of electroacupuncture(EA)on pain-ralated behaviors,morphology of hip-pocampus,concentrations of inflammatory cytokines and expression of ionized calcium binding adapter molecule 1(Iba-1)in dorsal horn of the spinal cord and the hippocampus,and brain-derived neurotrophic factor(BDNF)in hippocampus of rats with knee osteoarthritis(KOA),so as to explore the mechanism of EA in improving chronic pain of KOA.Methods Forty SD rats were randomly divided into blank group,saline group,model group and EA group,with 10rats in each group.Monosodium iodoacetate(MIA,80mg/mL,50μL)was injected into the left knee joint cavity of rats in the model group and EA group to establish the chro-nic pain model of KOA,while the same volume of normal saline was injected into the left knee joint cavity of rats in the saline group.Rats in the EA group received EA stimulation(2Hz/100Hz,1-2mA)at left“Yanglingquan”(GB34)and“Neixiyan”(EX-LE4)for 15min,14dafter MIA injection.The treatment was given once daily,5das 1session and 2sessions of treatment were required.Methanical withdrawl threshold(MWT)and weight-bearing capacity tests on left hind limbs were carried out 1d before,7d,14d,20dand 26dafter MIA injection.At the 27t h day,rats were sacrificed and HE staining was used to observe the morphology of hippocampal CA1area.Concentrations of interleukin(IL)-1βand tumor necrosis factor(TNF)-αin the left L3-L5 spinal dorsal horn and hippocampal CA1area were detected by ELISA,the expressions of Iba-1in the spinal dorsal horn and hippo-campal CA1area were detected by immunofluorescence,and the expression of BDNF in left hippocampal CA1area was detected by Western blot.Results The HE staining results of the hippocampal CA1area showed reduced number of neurons,unclear cell contour and boundary between nucleus and cytoplasm,and nuclear pyknosis in the model group,which was relatively milder in the EA group.Compared with the blank group,MWT and weight-bearing capacity of rats’left hind limbs,and expression of BDNF

关 键 词：膝关节骨关节炎 电针 疼痛 脊髓背角 海马 离子钙接头蛋白1 

分 类 号：R245.97[医药卫生—针灸推拿学]