电针通过抑制小胶质细胞激活拮抗吗啡镇痛耐受及相关机制  

Electroacupuncture antagonizes morphine analgesic tolerance by inhibiting microglia activation and related mechanisms

作　　者：赵昱 高菲 李岱[1] 吴冬梅[2] 徐礼鲜 ZHAO Yu;GAO Fei;LI Dai;WU Dongmei;XU Lixian(Department of Anesthesiology,Nanxishan Hospital of Guangxi Zhuang Autonomous Region,Guilin 541002,Guangxi Zhuang Autonomous Region,China;Department of Radiotherapy,Nanxishan Hospital of Guangxi Zhuang Autonomous Region,Guilin 541002,Guangxi Zhuang Autonomous Region,China;Department of Anesthesiology,School of Stomatology,Air Force Medical University,Xi’an 710032,Shaanxi,China)

机构地区：[1]广西壮族自治区南溪山医院麻醉科,桂林541002 [2]广西壮族自治区南溪山医院放射治疗科,桂林541002 [3]空军军医大学口腔医院麻醉科,西安710032

出　　处：《海军军医大学学报》2025年第3期411-417,共7页Academic Journal of Naval Medical University

基　　金：国家自然科学基金(81471265);广西医疗卫生适宜技术开发与推广应用项目(S2024074);桂林市技术应用与推广计划项目(20212E422580)。

摘　　要：目的观察电针对吗啡诱导的小胶质细胞激活和镇痛耐受的影响,探索电针治疗吗啡镇痛耐受的可能机制。方法将60只清洁级SD大鼠随机分为对照组、吗啡组、吗啡+电针组和吗啡+电针+集落刺激因子1(CSF1)组,每组15只。吗啡组、吗啡+电针组和吗啡+电针+CSF1组大鼠连续7 d鞘内注射吗啡,制备吗啡镇痛耐受模型;吗啡+电针组和吗啡+电针+CSF1组同时实施电针干预,选择疏密波,频率为2/100 Hz,以0.5、1.0和1.5 mA的刺激强度各刺激10 min,刺激“足三里”和“三阴交”穴,每天1次,连续7 d;吗啡+电针+CSF1组同时鞘内注射重组CSF1蛋白,连续7 d。采用机械缩足反射阈值(MWT)法观察电针对大鼠吗啡镇痛耐受的影响。7 d后处死大鼠,分离L4~6脊髓背角及背根神经节组织,采用蛋白质印迹法和qPCR法检测大鼠背根神经节及脊髓背角CSF1蛋白和mRNA表达,免疫荧光法检测脊髓背角小胶质细胞标志物离子化钙结合适配分子1(IBA-1)的表达,ELISA法检测脊髓IL-1β、IL-6和TNF-α表达。结果鞘内注射吗啡后,吗啡组大鼠最大镇痛效应百分率(%MPE)进行性降低,表明吗啡镇痛耐受模型构建成功;与吗啡组比较,吗啡+电针组大鼠在鞘内注射3、5和7 d时%MPE均增高(均P<0.05);与吗啡+电针组比较,吗啡+电针+CSF1组大鼠在鞘内注射3、5和7 d时%MPE均降低(均P<0.05)。与对照组比较,吗啡组大鼠背根神经节CSF1蛋白及mRNA表达、脊髓背角CSF1蛋白表达均增高(均P<0.05);与吗啡组比较,吗啡+电针组大鼠背根神经节CSF1蛋白及mRNA表达、脊髓背角CSF1蛋白表达均降低(均P<0.05);各组大鼠脊髓背角CSF1 mRNA表达差异均无统计学意义(均P>0.05)。与对照组比较,吗啡组大鼠脊髓背角IBA-1表达增高(P<0.05);与吗啡组比较,吗啡+电针组大鼠脊髓背角IBA-1表达降低(P<0.05);与吗啡+电针组比较,吗啡+电针+CSF1组大鼠脊髓背角IBA-1表达增高(P<0.05)。结论电针能够抑制大鼠脊髓背�Objective To observe the effects of electroacupuncture(EA)on morphine-induced microglia activation and analgesic tolerance,and explore the potential mechanism of EA in the treatment of morphine analgesic tolerance.Methods A total of 60 clean-grade SD rats were randomly assigned to control group,morphine group,morphine+EA group,and morphine+EA+colony-stimulating factor 1(CSF1)group,with 15 rats in each group.Morphine analgesic tolerance model was established by continuous 7-d intrathecal injection of morphine in the morphine,morphine+EA and morphine+EA+CSF1 groups.EA was given in the rats of morphine+EA and morphine+EA+CSF1 groups at“Zusanli”and“Sanyinjiao”acupoints,with dilatational wave,frequency of 2/100 Hz,stimulation intensities of 0.5,1.0,and 1.5 mA(10 min per intensity),once a day,for 7 consecutive days.Rats in morphine+EA+CSF1 group were given intrathecal injection with recombinant CSF1 protein for 7 consecutive days.The effect of EA on morphine analgesic tolerance in rats was observed by mechanical withdrawal threshold(MWT).After 7 d,the rats were sacrificed,and the L4-6 spinal dorsal horn and dorsal root ganglion tissues were isolated.The expression of CSF1 protein and mRNA in the dorsal root ganglia and spinal dorsal horn was detected by Western blotting and quantitative polymerase chain reaction.The expression of ionized calcium-binding adapter molecule 1(IBA-1),a marker of microglia in the spinal dorsal horn,was detected by immunofluorescence method,and the expression of interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF)-αin the spinal cord was detected by enzyme-linked immunosorbent assay(ELISA).Results After intrathecal injection of morphine,the percentage of maximal possible potential effect(%MPE)in the morphine group was decreased progressively,indicating that the morphine analgesic tolerance model was successfully constructed.Compared with the morphine group,the%MPE in the morphine+EA group was increased after intrathecal injection at 3,5 and 7 d(all P<0.05).Compared with the morphi

关 键 词：电针 吗啡镇痛耐受 小胶质细胞 集落刺激因子1 脊髓 背根神经节 

分 类 号：R24[医药卫生—中医临床基础]