Abstract
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.
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。
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References
GUO G X, ZHU Q G, SUN W Q, KONG L J, XU S D, ZHOU X, SONG P F, WU Z W, LÜ Z Z, FANG M. Research progress of immunological mechanism of Tuina intervention on lumbar disc herniation. Zhonghua Zhongyiyao Zazhi, 2019, 34(7): 3132–3135.
BENSON C A, FENRICH K K, OLSON K L, PATWA S, TAN A M. Dendritic spine dynamics after peripheral nerve injury: an intravital structural study. J Neurosci, 2020, 40(22): 4297–4308.
STRATTON H J, KHANNA R. Sculpting dendritic spines during initiation and maintenance of neuropathic pain. J Neurosci, 2020, 40(40): 7578–7589.
HUANG L E, GUO S H, THITISERANEE L, YANG Y, YAO Y X. N-methyl D-aspartate receptor subtype 2B antagonist, Ro 25-6981, attenuates neuropathic pain by inhibiting postsynaptic density 95 expression. Sci Rep, 2018, 8(1): 7848.
CHEN S J, JIANG Y, CHEN L C, CHEN J C, LIN Z G. Effect of massage on the expression of NMDAR2B in dorsal root ganglion and cornu dorsale medullae spinalis in rats with neuropathic pain. Guangming Zhongyi, 2019, 34(16): 2472–2474.
LIN Z G, WANG J Z, SONG P F, CHEN L C, LIN J, CHEN J C, JIANG Y, CHEN S J. Effects of Tuina pressing-kneading manipulation on NK1R and field potential of spinal dorsal horn in rats with lumbar disc herniation. Shizhen Guoyi Guoyao, 2020, 31(1): 253–256.
BENNETT G J, XIE Y K. A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain, 1988, 33(1): 87–107.
YU S G, XU B. Experimental Acupuncturology. Beijing: People’s Medical Publishing House, 2012.
NING L, MA L Q, WANG Z R, WANG Y W. Chronic constriction injury induced long-term changes in spontaneous membrane-potential oscillations in anterior cingulate cortical neurons in vivo. Pain Physician, 2013, 16(5): E577–E589.
WANG S D, ZHANG H, GUAN H Y, DONG B Q, TAO F J. Experimental study on effects of electroacupuncture on morphology of nerve roots and expressions of NK1 and NK2 in LDH rats. Zhonghua Zhongyiyao Xuekan, 2020, 38(12): 23–26.
TAO Y H, YAO B B, YU T Y, MA C, LU M Q, ZHANG L F, LÜ T T, WU S. Effect of Bo’fa method on the expression of IL-1β in peripheral serum and 5-HT2A in spinal cord of CCI model rats. Huanqiu Zhongyiyao, 2017, 10(8): 905–909.
WANG G C. Clinical study of bloodletting therapy plus acupuncture for lumbar disc herniation of blood stasis pattern. Shanghai Zhenjiu Zazhi, 2022, 41(3): 248–252.
DAI J F. Clinical observation of warm needling moxibustion plus lumbar traction for lumbar disc herniation. J Acupunct Tuina Sci, 2022, 20(4): 295–300.
LÜ L H, MAO L Y, LI J H, YANG C, WANG W W, WANG S, LIU D. Lever positioning manipulation combined with pulsed electric field on the analgesic effect of patients with lumbar disc herniation and its influence on IL-1β and TNF-α. Zhongguo Gushang, 2021, 34(8): 780–784.
LI L, YAO W L, ZHANG C H. The expression of A1 reactive astrocytes in spinal cord in rats with neuropathic pain. Zhongguo Tengtong Yixue Zazhi, 2020, 26(8): 579–583.
LIN Z G, CHEN S J, CHEN L C, CHEN J C, JIANG Y, WANG J Z. Analgesic effect of Tuina pressing and kneading method on neuropathic pain rats and its effect on SP in serum, dorsal root ganglia and spinal dorsal horn. Zhonghua Zhongyiyao Xuekan, 2019, 37(4): 877–880.
BENSON C A, REIMER M L, TAN A M. Dendritic spines in the spinal cord: live action pain. Neurosci Insights, 2020, 15: 2633105520951164.
BUSTOS F J, VARELA-NALLAR L, CAMPOS M, HENRIQUEZ B, PHILLIPS M, OPAZO C, AGUAYO L G, MONTECINO M, CONSTANTINE-PATON M, INTESTROSA N C, VAN ZUNDERT B. PSD95 suppresses dendritic arbor development in mature hippocampal neurons by occluding the clustering of NR2B-NMDA receptors. PLoS One, 2014, 9(4): e94037.
LAMBERT J T, HILL T C, PARK D K, CULP J H, ZITO K. Protracted and asynchronous accumulation of PSD95-family MAGUKs during maturation of nascent dendritic spines. Dev Neurobiol, 2017, 77(10): 1161–1174.
LIU Y, CUI X, SUN Y E, YANG X, NI K, ZHOU Y, MA Z, GU X. Intrathecal injection of the peptide myr-NR2B9c attenuates bone cancer pain via perturbing N-methyl-D-aspartate receptor-PSD-95 protein interactions in mice. Anesth Analg, 2014, 118(6): 1345–1354.
LÜ Z Z, ZHU Q G, KONG L J, CHENG Y B, GUO G X, ZHANG S P, ZHOU X, FANG M. Effects of manual loading on calcitonin gene-related peptide and nerve growth factor in rats with chronic low back pain. Zhongguo Gushang, 2021, 34(3): 282–287.
Acknowledgments
This work was supported by the Projects of National Natural Science Foundation of China (国家自然科学基金项目, No. 82105039, No. 82174523); Fujian Provincial Natural Science Foundation of China (福建省自然科学基金项目, No. 2020J01757, No. 2020J01758); Projects of Fujian Provincial Health Commission (福建省卫健委课题, No. 2020GGA070, No. 2020CXA052); Project of Education Department of Fujian Province (福建省教育厅课题, No. JAT200224); National TCM Innovation Key Talents Training Program [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].
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Zhang, H., Wang, B., Chen, S. et al. Exploring the effect of Tuina on the dendritic structure of spinal cord dorsal horn in rats with lumbar disc herniation based on NR2B/PSD-95 pathway. J. Acupunct. Tuina. Sci. 21, 129–136 (2023). https://doi.org/10.1007/s11726-023-1368-2
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DOI: https://doi.org/10.1007/s11726-023-1368-2
Keywords
- Tuina
- Massage
- Lumbar Vertebrae
- Point, Weizhong (BL40)
- Spinal Cord Dorsal Horn
- N-methyl-D-aspartate Receptor Subunit 2B
- Intervertebral Disc Displacement
- Rats