康复学报2025，Vol.35Issue(4)：399-408,10.DOI:10.3724/SP.J.1329.2025.04010
基于脊髓背角5-羟色胺及其受体调控途径探讨腕踝针治疗神经病理性疼痛的作用机制
Mechanism of Wrist-Ankle Acupuncture in Treating Neuropathic Pain Based on the Regula-tory Pathway of 5-Hydroxytryptamine and Its Receptors in the Spinal Dorsal Horn
摘要
Abstract
Objective To explore the mechanism by which wrist-ankle acupuncture(WAA)modulates neuropathic pain(NP)through the regulation of 5-hydroxytryptamine(5-HT)and its receptors in the spinal dorsal horn.Methods A total of 78 clean-grade male sprague-dawley rats were randomly divided into sham operation group,model group,WAA group,5-HT1AR antagonist group,5-HT2AR antagonist group,and 5-HT3R antagonist group after one week of acclimatization according to the random number table method,with 18,18,18,8,8,8 cases in each group respectively.Except for the sham operation group,chronic constriction in-jury(CCI)rat models were established in the other groups.From day 7 post-CCI,the WAA group,the 5-HT1AR antagonist group,the 5-HT2AR antagonist group and the 5-HT3R antagonist group received WAA treatment,once daily for 7 days.The sham operation group and the model group were similarly managed without acupuncture intervention.At the 9th,11th and 13th day after CCI oper-ation,the 5-HT1AR antagonist group,the 5-HT2AR antagonist group and the 5-HT3R antagonist group were administered intraperito-neal injections of WAY-100635,Ketanserin or Ondansetron antagonists respectively 30 minutes before WAA intervention.Before operation and on the 7th,14th day after operation,the pain behaviors[paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)]were evaluated.Immunofluorescence was used to detect tryptophan hydroxylase-2(TPH-2)expression levels in the dorsal raphe nucleus(DRN).Fluorescence microscopy was used to observe glial markers in the spinal dorsal horn,including ionized cal-cium-binding adapter molecule 1(Iba-1)and glial fibrillary acidic protein(GFAP).ELISA was used to detect expression levels of 5-HT and its receptors(5-HT1A,5-HT2A,5-HT3)in spinal dorsal horn;qRT-PCR was used to detect mRNA expression levels of inflam-matory cytokines interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in spinal dorsal horn.Results(1)Pain behavior:compared with the sham operation group,PWT and PWL in the model group on the 7th and 14th day after operation decreased sig-nificantly(P<0.05).Compared with the model group,PWT and PWL in the WAA group on the14th day after operation increased significantly(P<0.05).Compared with the WAA group,PWT and PWL in the 5-HT1AR antagonist group and the 5-HT3R antagonist group on the14th day after operation decreased significantly(P<0.05),while PWT in the 5-HT2AR antagonist group on the14th day after operation decreased significantly(P<0.05),but there was no significant difference in PWL(P>0.05).(2)TPH-2 and 5-HT levels:compared with the sham operation group,TPH-2 expression level in the DRN and 5-HT expression level in the spinal dorsal horn in the model group increased significantly(P<0.05);compared with the model group,TPH-2 expression level in the DRN and 5-HT expression level in the spinal dorsal horn in the WAA group decreased significantly(P<0.05).(3)Expression levels of 5-HT recep-tors:compared with the sham operation group,expression levels of 5-HT1AR and 5-HT3R receptors in the spinal dorsal horn in the model group decreased significantly(P<0.05),and expression level of 5-HT2AR receptor increased significantly(P<0.05).Com-pared with the model group,expression levels of 5-HT1AR and 5-HT3R receptors in the spinal dorsal horn in the WAA group in-creased significantly(P<0.05),and expression level of 5-HT2AR receptor decreased significantly(P<0.05).(4)Positive rates of Iba-1 and GFAP:compared with the sham operation group,the positive rates of Iba-1 and GFAP in the spinal dorsal horn in the model group increased significantly(P<0.05);compared with the model group,the positive rates of Iba-1 and GFAP in the WAA group de-creased significantly(P<0.05);compared with the WAA group,the positive rates of Iba-1 and GFAP in the 5-HT1AR antagonist group and the 5-HT3R antagonist group increased significantly(P<0.05),while there was no significant difference in Iba-1 and GFAP in the 5-HT2AR antagonist group(P>0.05).(5)mRNA transcription levels of IL-1β and TNF-α:compared with the sham op-eration group,the mRNA transcription levels of IL-1β and TNF-α in the dorsal horn of the spinal cord in the model group increased significantly(P<0.05);compared with the model group,the mRNA transcription levels of IL-1β and TNF-α in the dorsal horn of the spinal cord in the WAA group decreased significantly(P<0.05);compared with the WAA group,the mRNA transcription levels of IL-1β and TNF-α in the 5-HT1AR antagonist group and the 5-HT3R antagonist group increased significantly(P<0.05),while there was no significant difference in the mRNA transcription levels of IL-1β and TNF-α in the 5-HT2AR antagonist group(P>0.05).Con-clusion WAA can improve hyperalgesia of CCI model rats,and its mechanism may be related to reducing the activity of TPH-2 in DRN,regulating 5-HT and its receptors in the spinal dorsal horn,and inhibiting the activation of microglia to play an anti-inflam-matory role.关键词
神经病理性痛/腕踝针/中缝背核/脊髓背角/5-羟色胺Key words
neuropathic pain/wrist-ankle acupuncture/dorsal raphe nucleus/spinal dorsal horn/5-hydroxytryptamine引用本文复制引用
黄秋玲,洪益玲,龚德贵,兰艳艳,王志福,俞向梅..基于脊髓背角5-羟色胺及其受体调控途径探讨腕踝针治疗神经病理性疼痛的作用机制[J].康复学报,2025,35(4):399-408,10.基金项目
国家自然科学基金面上项目(82274658) （82274658）
福建省自然科学基金项目(2023J01333) （2023J01333）
福建省卫生健康委员会医学创新课题(2022CXA053) （2022CXA053）
福建省中青年教师教育科研项目(JAT220116) （JAT220116）
