电针调控背根神经节巨噬细胞浸润改善紫杉醇诱导神经病理痛的机制研究  

Mechanism of Electroacupuncture Mediated Macrophages Infiltration on Dorsal Root Ganglion to Alleviate Paclitaxel-Induced Neuropathic Pain

作　　者：向宏春 张虹 李静[1] 龙漫 李熳[3] 蔡国伟 XIANG Hongchun;ZHANG Hong;LI Jing;LONG Man;LI Man;CAI Guowei(Affiliated Union Hospital of Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei 430022,China;Hubei University of Science and Technology,Xianning,Hubei 437100,China;School of Basic Medicine,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei 430030,China)

机构地区：[1]华中科技大学同济医学院附属协和医院,湖北武汉430022 [2]湖北科技学院,湖北咸宁437100 [3]华中科技大学同济医学院基础医学院,湖北武汉430030

出　　处：《康复学报》2023年第6期521-527,共7页Rehabilitation Medicine

基　　金：国家自然科学基金项目(81774410,81973949);湖北省自然科学基金项目(2021CFB268)。

摘　　要：目的:观察电针双侧足三里穴对紫杉醇诱导神经病理痛小鼠模型背根神经节(DRG)巨噬细胞标记物CD68、单核细胞趋化蛋白-1(MCP-1)、白细胞介素-1β(IL-1β)和诱导型一氧化氮合酶(iNOS)表达的影响,探讨电针调控紫杉醇诱导神经病理痛的机制。方法:选择成年C57小鼠30只,采用随机数字表法分为对照组、模型组、电针组,每组10只。模型组、电针组在第1、3、5、7天分别通过腹腔注射2 mg/kg紫杉醇进行造模;对照组注射相同体积的溶媒。造模结束后(第9天),电针组给予电针双侧足三里穴治疗,频率15 Hz,强度1 mA,30 min/次,1次/d,共7 d。采用“up and down”法测定足底机械阈值(PWMT);采用苏木精和伊红(HE)染色观察各组DRG组织的病理特征;采用免疫荧光染色法检测DRG组织MCP-1和CD68免疫阳性面积百分比;采用Western blot法检测DRG组织CD68、MCP-1、iNOS和IL-1β蛋白表达水平;采用透射电镜观察DRG组织巨噬细胞超微结构。结果:①PWMT:与对照组同一时间点比较,模型组第8、11、13、15天PWMT明显降低(P<0.05);与模型组同一时间点比较,电针组第11、13、15天PWMT明显升高,差异具有统计学意义(P<0.05)。②DRG组织病理学特征:炎症细胞形态为近圆形,颜色为蓝紫色,对照组有少量的炎症细胞,模型组聚集浸润状态的炎症细胞数量增加,电针组聚集浸润状态的炎症细胞数量较少。③DRG组织MCP-1、CD68免疫阳性面积百分比:与对照组比较,模型组DRG组织MCP-1、CD68免疫阳性面积百分比明显升高(P<0.05);与模型组比较,电针组DRG组织MCP-1、CD68免疫阳性面积百分比明显降低(P<0.05)。④DRG组织MCP-1、CD68、IL-1β和iNOS蛋白表达水平:与对照组比较,模型组DRG组织MCP-1、CD68、IL-1β和iNOS蛋白表达水平明显升高(P<0.05);与模型组比较,电针组DRG组织MCP-1、CD68、IL-1β和iNOS蛋白表达水平明显下降(P<0.05)。⑤DRG组织巨噬细胞超微结构:对照组DRG组织巨�Objective:To observe the effects of electroacupuncture(EA)at bilateral Zusanli acupoint(ST 36)on the expressions of macrophage marker CD68,monocyte chemotactic protein-1(MCP-1),interleukin-1β(IL-1β)and inducible nitric oxide synthase(iNOS)in dorsal root ganglion(DRG)of mouse model with paclitaxel-induced neuropathic pain,and to explore the mechanism of EA in regulating paclitaxel-induced neuropathic pain.Methods:A total of 30 adult C57 mice were randomly divided into control group,model group and EA group,with 10 mice in each group.The model group and the EA group received intraperitoneal injection of 2 mg/kg paclitaxel on the 1st,3rd,5th and 7th day,respectively,while the control group was injected with the same volume of solvent.After injection(at the 9th day),the EA group received electroacupuncture at bilateral Zusanli acupoint(ST 36)with frequency of 15 Hz,intensity of 1 mA,30 minutes a time,once a day,lasting for seven days.The"up and down"method was used to measure paw withdrawal mechanical threshold(PWMT);the hematoxylin-eosin(HE)staining was used to observe pathological characteristics of DRG tissues;immunofluorescence staining was used to detect the percentage of MCP-1 and CD68 immunopositive area in DRG tissues;Western blot method was used to detect the protein expression level of CD68,MCP-1,iNOS and IL-1βin DRG tissues,and transmission electron microscope was used to observe the ultrastructure of macrophages in DRG tissues.Results:(1)PWMT:compared with the control group at the same time,PWMT in the model group decreased significantly at the 8th,11th,13th and 15th days(P<0.05);compared with the model group at the same time,PWMT in the EA group increased significantly at the 11th,13th and 15th days,and the differences were statistically significant(P<0.05).(2)Histopathological features of DRG tissues:the inflammatory cells are nearly round in shape and blue-purple in color.There were a small number of inflammatory cells in the control group,the number of inflammatory cells in the aggregating and infi

关 键 词：神经病理痛 电针 紫杉醇 足三里穴 背根神经节 巨噬细胞 

分 类 号：R730.53[医药卫生—肿瘤]