基于SNAP25/VGLUT2通路探讨推拿对坐骨神经慢性压迫损伤大鼠脊髓背角谷氨酸含量及突触超微结构的影响  

Exploration on the Effects of Tuina on Glutamate Content and Synaptic Ultrastructure in Spinal Dorsal Horn of Rats with Chronic Sciatic Nerve Compression Injury Based on the SNAP25/VGLUT2 Pathway

作　　者：蒋晶晶 黄丽梅 黄红叶 蔡恒昌 张幻真 陈乐春 陈水金 伍诗烨 林惠 林志刚 JIANG Jingjing;HUANG Limei;HUANG Hongye;CAI Hengchang;ZHANG Huanzhen;CHEN Lechun;CHEN Shuijin;WU Shiye;LIN Hui;LIN Zhigang(Rehabilitation Hospital Affiliated to Fujian University of Chinese Medicine,Fuzhou 350003,China;Fujian Provincial Key Laboratory of Rehabilitation Technology,Fuzhou 350003,China;Fujian University of Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]福建中医药大学附属康复医院,福建福州350003 [2]福建省康复技术重点实验室,福建福州350003 [3]福建中医药大学,福建福州350122

出　　处：《中国中医药信息杂志》2025年第4期113-119,共7页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金(82205303、82105039);国家自然科学基金面上项目(82174523);福建省自然科学基金杰青项目(2023J06037);福建省自然科学基金面上项目(2023J01878);福建省卫健委中青年骨干科研项目(2021GGA060);福建省卫生健康中青年领军人才研修项目(2023年)。

摘　　要：目的观察推拿对坐骨神经慢性压迫损伤大鼠脊髓背角谷氨酸含量及突触超微结构的影响,探讨推拿调控SNAP25/VGLUT2通路缓解腰椎间盘突出症的潜在机制。方法采用坐骨神经慢性压迫损伤模拟腰椎间盘突出症神经病理性疼痛,将24只SD大鼠随机分为空白组、模型组和推拿组,每组8只。造模后第4日起,推拿组按揉法干预,10 min/次,1次/d,连续14 d。检测造模前及造模后第4、10、14、17日大鼠机械缩足阈值(PWT)、热痛阈值(PWL),取造模侧脊髓组织,透射电镜观察脊髓背角神经元突触超微结构,免疫荧光染色检测脊髓背角N甲基-D-天冬氨酸受体(NR)2A表达,Western blot检测脊髓背角突触相关蛋白25(SNAP25)蛋白表达,免疫组化法检测脊髓背角囊泡谷氨酸转运体2(VGLUT2)表达,ELISA检测脊髓背角谷氨酸含量。结果与空白组比较,模型组大鼠造模后第4、10、14、17日PWT、PWL明显降低(P<0.001),脊髓背角突触前膜内囊泡聚集,突触后致密区面积增大,突触间隙增大,脊髓背角NR2A、SNAP25、VGLUT2蛋白表达明显升高(P<0.05,P<0.001),谷氨酸含量明显升高(P<0.001);与模型组比较,推拿组大鼠造模后第10、14、17日PWT、PWL明显升高(P<0.001),突触囊泡均匀分布,突触后致密区面积减小,突触间隙减小,脊髓背角NR2A、SNAP25、VGLUT2蛋白表达明显降低(P<0.05,P<0.001),谷氨酸含量明显降低(P<0.01)。结论推拿可能通过调控脊髓背角SNAP25/VGLUT2通路影响谷氨酸含量,改善神经元突触超微结构,对腰椎间盘突出症起到镇痛作用。Objective To observe the effect of tuina on glutamate content and synaptic ultrastructure in spinal dorsal horn of rats with chronic sciatic nerve compression injury;To explore the potential mechanism of tuina regulation of the SNAP25/VGLUT2 pathway in alleviating lumbar disc herniation.Methods A chronic sciatic nerve compression injury model was used to simulate neuropathic pain in lumbar disc herniation.24 SD rats were randomly divided into blank group,model group and tuina group,with 8 rats in each group.From the 4th day after modeling,the tuina group was intervened with the tuina method for 10 minutes once a day for 14 consecutive days.The paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)of rats in each group on the day before modeling,and the 4th,10th,14th and 17th days after modeling were detected.The spinal cord tissue of the modeling side was taken,synaptic ultrastructure of spinal dorsal horn neurons was observed using transmission electron microscopy,immunofluorescence staining was used to detect the expression of NR2A in the spinal dorsal horn,Western blot was used to detect the expression of SNAP25 protein in the spinal dorsal horn,immunohistochemistry was used to detect the expression of VGLUT2 in the spinal dorsal horn,ELISA was used to detect the content of glutamate in the spinal dorsal horn.Results Compared with the blank group,PWT and PWL of the model group were significantly reduced on the 4th,10th,14th and 17th days after modeling(P<0.001),with accumulation of vesicles in the presynaptic membrane of the dorsal horn of the spinal cord,increase in the area of the postsynaptic dense zone,and enlargement of the synaptic cleft,while the protein expressions of NR2A,SNAP25 and VGLUT2 in the spinal dorsal horn increased(P<0.05,P<0.001),and the content of glutamate increased(P<0.001).Compared with the model group,PWT and PWL of the tuina group rats significantly increased on the 10th,14th and 17th days after modeling(P<0.001),synaptic vesicles were evenly distributed,the area of the postsyna

关 键 词：腰椎间盘突出症 推拿 脊髓背角 SNAP25/VGLUT2通路 谷氨酸 突触超微结构 大鼠 

分 类 号：R247[医药卫生—中医临床基础]