摘要
目的:通过观察N-甲基-D-天门冬氨酸受体2B亚基(NR2B)/突触后致密蛋白95(PSD-95)通路对腰椎间盘突出症大鼠脊髓背角树突结构的影响探讨推拿镇痛的作用机制.方法:将50只Sprague-Dawley大鼠随机分为空白组、模型组、推拿组、阻断剂组和阻断剂+推拿组,采用坐骨神经结扎法制备坐骨神经慢性压迫性损伤(CCI)模型,推拿组和阻断剂+推拿组在造模后第4天起每日实施推拿干预,阻断剂组和阻断剂+推拿组在造模后第4天起每日鞘内注射NR2B阻断剂(MK-801).采用自发痛评分观察各组大鼠的疼痛行为.连续干预14 d后运用免疫组化法检测各组大鼠脊髓背角NR2B及下游PSD-95蛋白的表达水平,高尔基染色法观察大鼠脊髓背角树突的结构变化.结果:相较于空白组,模型组和推拿组大鼠在CCI术后自发痛程度均升高(P<0.01),推拿干预后,推拿组大鼠疼痛程度较模型组减轻(P<0.05).阻断NR2B后,阻断剂组和阻断剂+推拿组大鼠疼痛程度均较模型组减轻(P<0.05).相较于空白组,模型组大鼠NR2B和PSD-95蛋白含量明显升高(P<0.01);脊髓背角树突总分叉数增加(P<0.01),总长度变长(P<0.01).相较于模型组,推拿组大鼠NR2B和PSD-95蛋白含量显著下降(P<0.01);脊髓背角树突总分叉数减少(P<0.01),总长度缩短(P<0.01).阻断NR2B后,阻断剂组和阻断剂+推拿组的NR2B及下游PSD-95蛋白的表达水平较模型组均显著降低(P<0.01);脊髓背角树突总分叉数明显减少(P<0.01),总长度显著缩短(P<0.01).结论:推拿可通过重塑腰椎间盘突出症大鼠脊髓背角树突结构发挥镇痛作用,其机制可能与抑制NR2B/PSD-95信号通路的表达有关.
Objective To investigate the analgesic mechanism of Tuina(Chinese therapeutic massage)by observing the effect of the N-methyl-D-aspartate receptor subunit 2B(NR2B)/postsynaptic density-95(PSD-95)pathway on the dendritic structure of spinal cord dorsal horn in rats with lumbar disc herniation.Methods Fifty Sprague-Dawley rats were randomly divided into a blank group,a model group,a Tuina group,a blocker agent group,and a blocker agent+Tuina group.The sciatic nerve chronic constriction injury(CCI)model was prepared by the sciatic nerve ligation method.From the 4th day after modeling,rats in the Tuina group and the blocker agent+Tuina group were subject to daily Tuina intervention,and those in the blocker agent group and the blocker agent+Tuina group were daily intrathecally injected with NR2B blocker agent(MK-801).The spontaneous pain score was used to observe the pain behavior of all rats.The expression levels of NR2B and downstream PSD-95 were measured by immunohistochemistry,and the dendritic structure changes were observed by Golgi staining for rat spinal cord dorsal horn after 14 d of continuous intervention.Results Compared with the blank group,the degree of rat spontaneous pain after CCI was elevated in both the model and the Tuina groups(P<0.01)and was reduced in the Tuina group after the Tuina intervention compared with the model group(P<0.05).Compared with the model group,the rat spontaneous pain level after blocking NR2B was reduced in both the blocker agent group and the blocker agent+Tuina group(P<0.05).The NR2B and PSD-95 protein levels were significantly higher in the model group compared with the blank group(P<0.01);the total number of dendritic branches was increased(P<0.01),and the total dendritic length became longer(P<0.01)in the spinal cord dorsal horn.The rat NR2B and PSD-95 protein levels were significantly decreased in the Tuina group compared with the model group(P<0.01);the total dendritic branch number was reduced(P<0.01)and the total length was shortened(P<0.01)in the spinal cord dorsal horn.After blocking NR2B,the expression levels of NR2B and downstream PSD-95 protein were significantly lower in both the blocker agent group and the blocker agent+Tuina group compared to the model group(P<0.01).The total branch number was significantly reduced(P<0.01),and the total length was significantly shortened(P<0.01)of the dendrites in the spinal cord dorsal horn.Conclusion Tuina may exert an analgesic effect by remodeling the dendritic structure in the spinal cord dorsal horn in rats with lumbar disc herniation,and its mechanism may be related to the inhibition of NR2B/PSD-95 signaling pathway.
作者
张幻真
王冰倩
陈水金
陈乐春
蒋晶晶
江煜
陈进城
黄红叶
方佳钰
曾维铨
林志刚
ZHANG Huanzhen;WANG Bingqian;CHEN Shuijin;CHEN Lechun;JIANG Jingjing;JIANG Yu;CHEN Jincheng;HUANG Hongye;FANG Jiayu;ZENG Weiquan;LIN Zhigang(Rehabilitation Hospital Affiliated to Fujian University of Traditional Chinese Medicine,Fuzhou,350003,China;Fujian Key Laboratory of Rehabilitation Technology,Fuzhou,350003,China;Fujian University of Traditional Chinese Medicine,Fuzhou,350122,China)
基金
国家自然科学基金项目,No.82105039,No.82174523
福建省自然科学基金项目,No.2020J01757,No.2020J01758
福建省卫健委课题,No.2020GGA070,No.2020CXA052
福建省教育厅课题,No.JAT200224
全国中医药创新骨干人才培训项目,国中医药人教函(2019)128号.
关键词
推拿
按摩
腰椎
穴
委中
脊髓背角
N-甲基-D-天门冬氨酸受体2B亚基
椎间盘移位
大鼠
Tuina
Massage
Lumbar Vertebrae
Point,Weizhong(BL40)
Spinal Cord Dorsal Horn
N-methyl-D-aspartate Receptor Subunit 2B
Intervertebral Disc Displacement
Rats
