电针对慢性神经痛大鼠穴区高迁移率族蛋白1及其受体CD 24表达的影响  

High Mobility Group Box 1/CD 24 Receptor/β-EP Signaling in“Zusanli”(ST 36)Region Contributes to Electroacupuncture Analgesia in Rats with Neuropathic Pain

作　　者：高永辉[1] 王俊英[1] 谭连红[1] 端木程琳 乔丽娜[1] 韩焱晶[1] 张建梁[1] 刘俊岭[1] GAO Yong-hui;WANG Jun-ying;TAN Lian-hong;DUANMU Cheng lin;QIAO Li-na;HAN Yan-jing;ZHANG Jian liang;LIU Jun-ling(Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院针灸研究所,北京100700

出　　处：《针刺研究》2018年第9期537-542,共6页Acupuncture Research

基　　金：国家重点基础研究发展计划"973"中医专项(No.2013CB 531904);国家自然科学基金项目(No.81473779)

摘　　要：目的:观察电针干预治疗慢性压迫性损伤(CCI)大鼠过程中,大鼠"足三里"穴区局部高迁移率族蛋白1(HMGB 1)及其受体CD 24表达及β-内啡肽(β-EP)含量的变化,探讨穴区电针刺激缓解慢性神经痛的机制。方法:Wistar大鼠随机分为正常对照组、CCI模型组、电针组,每组10只。正常对照组大鼠行假手术,其余两组结扎坐骨神经造成慢性神经痛模型。手术后1周开始测定大鼠双侧热刺激缩腿反应的潜伏期(PWL)。电针组于CCI术后8d电针双侧"足三里""阳陵泉"穴,连续5d。电针结束后用免疫沉淀法检测"足三里"穴区乙酰化HMGB 1蛋白表达,用蛋白免疫印迹、荧光定量PCR分别检测HMGB 1和Toll样受体4(TLR 4)蛋白、CD 24mRNA的表达,用酶联免疫吸附(ELISA)法检测穴位局部组织β-EP的含量,并在穴位局部注射CD 24的中和抗体,观察阻断HMGB 1-CD 24通路对电针效应的影响。结果:与正常对照组比较,CCI后各组动物双侧PWL差值明显增加(P<0.05);与CCI模型组比较,电针组动物的痛觉过敏明显减轻(P<0.05)。CCI后穴位局部的HMGB 1和TLR 4蛋白表达比正常对照组明显升高(P<0.05);电针后,和CCI模型组比较,HMGB 1乙酰化程度增高,CD 24mRNA表达升高,同时穴位局部β-EP含量显著升高(P<0.05),但是HMGB 1及TLR 4蛋白表达未见明显变化(P>0.05)。穴位局部注射CD 24的中和抗体后,电针升高局部β-EP含量、抑制动物痛觉过敏的作用明显减弱(P<0.05)。结论:电针"足三里""阳陵泉"穴可缓解CCI大鼠的神经病理性疼痛,其效应与穴位局部HMGB 1活性及其受体CD 24表达升高导致的局部β-EP含量增加有关。Objective To observe the effect of electroacupuncture （EA） on the expression of high mobility group box 1 （HMGB 1） and its receptor cD 24 proteins and 13-endorphin （IB-EP） content in ＂Zusanli＂ （ST 36） region in rats with chronic con- striction injury （Ccl）, so as to explore its mechanisms underlying pain relief. Methods Thirty male Wistar rats were rando- mized into control, COl model and EA groups （n = 10 rats in each）. The neuropathic pain model was established by ligature of the left sciatic nerve to induce CCI in the model and EA groups, and sham operation was performed in rats of the control group. Paw with drawal latency （PWL, thermal pain threshold） of the bilateral hind-limbs was detected by using an algesia-detector. Eight days after COl operation, EA was applied to bilateral ＂Zusanli＂ （ST 36） and ＂Yanglingquan＂ （GB 34） for 30 min, once daily for 5 days. The acetylated-HMGB 1 expression was determined by immunoprecipitation, and the expression of HMGB 1 and toll like receptor 4 （TLR 4） proteins and CD 24 mRNA were detected using Western blot and fluorescent quantitative real time-PCR, respectively, and the content of IB-EP in the acupoint region was assayed by enzyme linked immunosorbent assay （ELISA）. Anti-cD 24 neutralizing antibody （200 μL, 100 μg/mL） was injected into ST 36 region once daily for 3 days for verifying the involvement of HMGB 1/CD 24 signaling in EA analgesia. Results Compared with the control group, the bilateral PWL difference values in the other two groups were significantly increased （P〈0.05）, meaning an occurrence of hyperalgesia after CCI. In comparison with the Cci model group, the hyperalgesia in the EA group was obviously decreased （P〈0.05）. After CCI, the expression levels of HMGB 1 and TLR 4 proteins were considerably increased compared with those in the control group （P〈0.05）. After 5-times＇ EA, the acetylated-HMGB 1, the expression of CD 24 mRNA, and the content of β-EP were notably up-regula

关 键 词：慢性神经痛 针刺镇痛 高迁移率族蛋白1 CD 24 Toll样受体4 Β-内啡肽 

分 类 号：R246.2[医药卫生—针灸推拿学]