火针对化疗所致神经病理性疼痛大鼠外周敏化的镇痛作用机制  

Mechanism of analgesic effect of fire needle on peripheral sensitization in rats with neuropathic pain induced by chemotherapy

作　　者：曾婧纯[1] 陈悦轩 李晶晶[3] 王鑫栋 肖茜 张瑞琳 纪铠松 陆丽明[5] ZENG Jing-chun;CHEN Yue-xuan;LI Jing-jing;WANG Xin-dong;XIAO Xi;ZHANG Rui-lin;JI Kai-song;LU Li-ming(Rehabilitation Center of The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510000,China;First Clinical Medical College,Guangzhou University of Chinese Medicine,Guangzhou 510000;Department of Acupuncture,Bao’an District Hospital of Traditional Chinese Medicine,Shenzhen 518000,Guangdong Province;Henan Zhang Zhongjing College of Traditional Chinese Medicine,Nanyang 473004,Henan Province;South China Research Center for Acupuncture and Moxibustion,Guangzhou University of Chinese Medicine,Guangzhou 510000)

机构地区：[1]广州中医药大学第一附属医院康复中心,广州510000 [2]广州中医药大学第一临床医学院,广州510000 [3]深圳市宝安区中医院针灸科,广东深圳518000 [4]河南张仲景国医国药学院,河南南阳473004 [5]广州中医药大学华南针灸研究中心,广州510000

出　　处：《针刺研究》2023年第11期1095-1102,共8页Acupuncture Research

基　　金：广东省中医药局项目(No.20211340);广东省基础与应用基础研究基金青年项目(No.2019A1515110371);广州中医药大学第一附属医院创新强院青年项目(No.2019QN23);广州市科技创新局项目(No.202201011206)。

摘　　要：目的:从外周敏化角度探讨火针治疗奥沙利铂诱导的神经病理性疼痛(NP)大鼠外周敏化的镇痛机制。方法:SD大鼠随机分为正常组(n=6)、模型组(n=6)、火针组(n=6)和药物组(n=6)。采用腹腔注射奥沙利铂(4 mg/kg)建立NP大鼠模型。造模后的第1、3、5天(即实验的第24、26、28天),火针组行火针L4—L6“夹脊”治疗,药物组行普瑞巴林(100 mg/kg)腹腔注射治疗。Von Frey丝检测大鼠缩足阈值(PWT)代表机械痛阈值,ELISA法检测大鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、趋化因子12(CXCL12)含量,HE染色法观察大鼠穴区皮肤病理形态,甲苯胺蓝染色法观察各组大鼠穴区皮肤肥大细胞数量,免疫组织化学法检测大鼠穴区皮肤类胰蛋白酶(TPS)、瞬时受体电位香草酸通道1(TRPV1)、蛋白酶活化受体2(PAR2)阳性表达,Western blot法检测大鼠背根神经节(DRG)中TRPV1和PAR2蛋白的表达。结果:与正常组相比,模型组大鼠造模后的PWT降低(P<0.05),血清IL-6、TNF-α、CXCL12含量升高(P<0.05),穴区皮肤肥大细胞数量增加(P<0.05),穴区皮肤TPS、TRPV1和PAR2蛋白阳性表达升高(P<0.05)。与模型组相比,火针组和药物组大鼠干预后的PWT升高(P<0.05),血清IL-6、TNF-α、CXCL12含量,穴区皮肤TPS、TRPV1、PAR2蛋白阳性表达降低(P<0.05);药物组DRG中TRPV1和PAR2蛋白表达量降低(P<0.05)。HE染色结果显示,模型组大鼠穴区表皮层增厚,细胞排列层次紊乱,细胞间水肿明显,真皮层内可见大量炎性细胞浸润;药物组、火针组穴区表皮层较薄,细胞排列层次较清晰,组织水肿和炎性细胞浸润情况均较模型组减轻。结论:火针可以改善奥沙利铂诱导的NP大鼠的机械痛阈、降低外周炎性因子的水平,该作用可能与抑制肥大细胞活化,抑制穴区局部TPS、TRPV1和PAR2蛋白的表达有关。Objective This study aims to explore the analgesic mechanism of fire needle on peripheral sensiti⁃zation in rats with neuropathic pain(NP)induced by oxaliplatin,so as to investigate its mechanism in improving peripheral sensitization.Methods Male SD rats aged 8 weeks were randomly divided into 4 groups:normal group(n=6),model group(n=6),fire needle group(n=6),and medication group(n=6).NP rat model was established by intraperito⁃neal injection of oxaliplatin(4 mg/kg)on days 1,2,8,9,15,16,22,and 23.For rats in the fire needle group,fire needle treatment was performed at the“Jiaji”(EX-B2)acupoints of the L4—L6 segments on days 24,26,and 28,ie.1 day,3 and 5 days after modeling.The medication group received intraperitoneal injection of pregabalin(100 mg/kg).Mechanical pain thresholds of the rats were measured before modeling,after modeling and intervention.Serum con⁃tents of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and chemokine ligand 12(CXCL12)were detected by ELISA.Skin histopathology changes in the acupoint area were observed using HE staining.The number of mast cells in the skin of the acupoints was observed using toluidine blue staining.Immunohistochemical staining was performed to detect the postive expressions of transient receptor potential vanilloid 1(TRPV1),protease-activated receptor 2(PAR2)and tryptase(TPS)in the skin of the acupoint area.Western blot was used to detect the protein expressions of TRPV1 and PAR2 in the dorsal root ganglia(DRG).Results Compared with the normal group,the model group had de⁃creased paw withdrawal threshold(PWT)after modeling(P<0.05),increased serum contents of IL-6,TNF-α,and CXCL12(P<0.05),increased number of mast cells in the acupoint area(P<0.05),and increased positive protein ex⁃pressions of TPS,TRPV1,and PAR2 in the skin of the acupoint area(P<0.05).Compared with the model group,the fire needle group and medication group had increased PWT after intervention(P<0.05),decreased serum contents of IL-6,TNF-α,and CXCL12,and postive protein expressio

关 键 词：神经病理性疼痛 火针 瞬时受体电位香草酸通道1/蛋白酶活化受体2 奥沙利铂 肥大细胞 神经炎性反应 

分 类 号：R245.319[医药卫生—针灸推拿学]