坐骨神经胞外核苷酸酶在针刺缓解大鼠踝关节炎性痛中的作用  

Involvement of extracellular nucleotidases in sciatic nerve in acupuncture alleviating inflammatory pain of ankle arthritis in rats

作　　者：徐靖雯 李雨佳 汤思琦 沈诞 林捷 汪丽娜[1,3] 沈雪勇 XU Jingwen;LI Yujia;TANG Siqi;SHEN Dan;LIN Jie;WANG Lina;SHEN Xueyong(School of Acupuncture-Moxibustion and Tuina,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Department of Traditional Chinese Medicine,Chinese People's Liberation Army Naval Medical University,Shanghai 200433,China;Shanghai Key Laboratory of Acupuncture Mechanism and Acupoint Function,Shanghai 200433,China;Shanghai Research Centre for Acupuncture and Meridians,Shanghai 201203,China)

机构地区：[1]上海中医药大学针灸推拿学院,上海201203 [2]中国人民解放军海军军医大学中医系,上海200433 [3]上海市针灸机制与穴位功能重点实验室,上海200433 [4]上海市针灸经络研究中心,上海201203

出　　处：《上海中医药杂志》2024年第2期23-30,共8页Shanghai Journal of Traditional Chinese Medicine

基　　金：上海市自然科学基金项目(22ZR1461500);上海中医药大学预算内项目(2021LK098)。

摘　　要：目的 探讨坐骨神经胞外核苷酸酶[CD39(NTPDase1)]在针刺缓解大鼠踝关节炎性痛中的作用。方法 将54只雄性大鼠随机分为9组,即空白组、踝关节炎性痛模型组、针刺组、坐骨神经腺苷三磷酸(ATP)水解酶抑制(ARL67156注射)+针刺组、坐骨神经CD39上调(三磷酸腺苷双磷酸酶注射)组、坐骨神经CD39特异性抑制(噻氯匹啶注射)+针刺组、坐骨神经CD73特异性抑制(α,β-亚甲基腺苷5′-二磷酸钠盐注射)+针刺组、穴区CD39上调(三磷酸腺苷双磷酸酶注射)组、穴区CD39特异性抑制(噻氯匹啶注射)+针刺组,每组6只。除空白组外,其余各组大鼠均采用注射完全弗氏佐剂建立佐剂性关节炎疼痛模型;造模48 h后,需针刺组大鼠针刺患侧“足三里”穴20 min,需注射药物组于针刺前20 min进行局部相关试剂注射。以大鼠患侧足底机械痛阈值和热痛阈值作为疼痛评价指标;采用实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法和Western blot法检测坐骨神经以及“足三里”穴胞外核苷酸酶的表达。结果 (1)调节坐骨神经ATP水解酶活性:与空白组比较,踝关节炎性痛模型组和坐骨神经ATP水解酶抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05);与踝关节炎性痛模型组比较,针刺组热痛阈值和机械痛阈值明显上升(P<0.05)。(2)调节坐骨神经CD39活性:与空白组比较,坐骨神经CD39特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05);与踝关节炎性痛模型组比较,针刺组和坐骨神经CD39上调组热痛阈值和机械痛阈值明显上升(P<0.05);与针刺组比较,坐骨神经CD39特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05)。(3)调节穴区CD39活性:与空白组比较,穴区CD39特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05);与踝关节炎性痛模型组比较,针刺组和穴区CD39上调组热痛阈值和机械痛阈值明显上升(P<0.05);与针刺组比较,�Objective To investigate the role of extracellular nucleotidase[CD39(NTPDase1)]in acupuncture alleviating inflammatory pain of ankle arthritis(AA)in rats.Methods Fifty-four male rats were randomly divided into 9 groups:control group,AA model group,acupuncture group,sciatic nerve ATP hydrolase inhibitor(ARL67156 injection)+acupuncture group,sciatic nerve CD39 upregulation(Apyrase injection)group,sciatic nerve CD39 enzyme-specific inhibitor(Ticlopidine injection)+acupuncture group,sciatic nerve CD73 enzyme-specific inhibitor(AMP-CP injection)+acupuncture group,acupoint region CD39 upregulation(Apyrase injection)group,acupoint region CD39 enzyme-specific inhibitor(Ticlopidine injection)+acupuncture group,with 6 rats in each group.Except for rats in the control group,the remaining rats in each group were injected with complete Freund's adjuvant to establish an adjuvant-induced arthritis pain model.After 48 h of modeling,rats in the acupuncture group were needled at the Zusanli(ST36)on the affected side for 20 min.Each pretreatment group performed local administration of relevant reagents 20 min before acupuncture.The mechanical pain threshold(MPT)and thermal pain threshold(TPT)of the planta pedis of the affected rat paw were used to evaluate pain levels.Real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot were used to detect the expression level of CD39 in the sciatic nerve and Zusanli(ST36)point.Results①Regulating ATP hydrolase activity in sciatic nerve:Compared with the control group,the AA model group and the sciatic nerve ATP hydrolase inhibitor+acupuncture group showed a significant decrease in TPT and MPT(P<0.05);compared with the AA model group,the acupuncture group showed a significant increase in TPT and MPT(P<0.05).②Regulating CD39 activity in sciatic nerve:Compared with the control group,the sciatic nerve CD39 enzyme-specific inhibitor+acupuncture group showed a significant decrease in TPT and MPT(P<0.05);compared with the AA model group,the acupuncture group and the sciatic nerve

关 键 词：针刺镇痛 炎症反应 关节疼痛 足三里 坐骨神经 胞外核苷酸酶 作用机制 

分 类 号：R245[医药卫生—针灸推拿学]