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.

Objective To investigate the effects of abdominal tuina on the expression of PI3K and N-methyl-D-aspartate receptor(NMDAR)subunit NR1 in spinal dorsal horn and the morphology of spinal dorsal horn neurons in ulcerative colitis(UC)rats;To explore its mechanism of action in treating UC.Methods Totally 36 SD rats were randomly divided into normal group,model group,abdominal tuina group,mesalazine group,PI3K stimulation group and PI3K stimulation + abdominal tuina group,with 6 rats in each group.The UC model in rats was simulated by drinking dextran sulfate solution freely.The abdominal tuina group and the PI3K stimulation + abdominal tuina group were given abdominal tuina intervention,the mesalazine group was given mesalazine solution for gavage,and the PI3K stimulation group and PI3K stimulation + abdominal tuina group were given intrathecal injection of PI3K agonist,once a day,for consecutive 15 days.Abdominal withdrawal reflex(AWR)score and acetic acid twist were used to observe the abdominal pain symptoms in rats.The expression of PI3K and NR1 in spinal dorsal horn were detected by immunofluorescence staining and Western blot,and the morphological changes of spinal dorsal horn neurons were observed by Nissl staining.Results Compared with the normal group,AWR score and twisting times of rats in model group significantly increased(P<0.01),the expression of PI3K and NR1 protein in spinal dorsal horn significantly increased(P<0.05,P<0.01),the morphology of spinal dorsal horn neurons was disordered,forming a large number of vacuolar like structures,and the Nissl body structure was fuzzy and incomplete.Compared with the model group,AWR scores and twisting times of abdominal tuina group and mesalazine group significantly decreased(P<0.05,P<0.01),and the expression of PI3K and NR1 protein significantly decreased(P<0.05,P<0.01),the edema of spinal dorsal horn neurons was milder,with fewer vacuolar changes and an increase in the number of Nissl bodies;AWR scores and twisting times of PI3K stimulation group and PI3K stimulation + abdominal tuina group significantly increased(P<0.05,P<0.01),and the expressions of PI3K and NR1 protein increased(P<0.05,P<0.01),a large number of neurons underwent pyknosis and necrosis,and the number of Nissl bodies decreased,even dissolving and disappearing.Conclusion Abdominal tuina can effectively improve the symptoms of abdominal pain in UC model rats,and its mechanism may be related to inhibiting the expression of PI3K and NR1 in spinal dorsal horn and improving the morphology of spinal dorsal horn neurons.

ObjectiveTo explore the possible mechanism of Tuina at Weizhong （BL 40） for relieving sciatica from the perspective of hippocampal synaptic plasticity. MethodsSixty SD rats were randomly divided into sham operation group， model group， Tuina group， MK-801 group， MK-801 plus Tuina group， 12 rats in each group. After lateral ventricular cannulation， rats model with chronic compression injury of the right sciatic nerve were prepared in all groups except the sham operation group. On day 4 after modelling， rats in the Tuina group start Tuina at Weizhong （BL 40） for 10 mins once a day for a total of 14 days； rats in the MK-801 group started injecting with 0.25 μg/μl of the N-methyl-D-aspartate receptor 2B （NR2B） blocker， dizocycline （MK-801）， 0.5 μl of which was administered daily in the lateral ventricle for 14 days. Rats in the MK-801 plus Tuina group underwent Tuina after 30 mins when completing MK-801 injection in the lateral ventricle， in the same way as above； rats in the model group and the sham operation group did not undergo any intervention. Spontaneous pain behaviour scores and paw withdraw thresholds （PWTs） were examined on day 1 （base value） before modelling and on day 4， 10， 14 and 18 after modelling； and on day 19， the brain tissues of the rats in each group were sampled and the number and morphology of the Nysted-positive cells in the hippocampal CA3 region were observed using Nysted staining； and the number of synapses， the thickness of postsynaptic dense material， the length of active band and the curvature of synaptic interface in hippocampal CA3 region were observed by transmission electron microscopy； and the expression of synapse-associated proteins NR2B and postsynaptic density protein-95 （PSD95） in hippocampal CA3 was detected by immunofluorescence staining and immunoblotting. ResultsCompared with the same time in the sham operation group， spontaneous pain scores significantly increased and PWTs decreased on day 4， 10， 14， and 18 after modelling in the model group （P＜0.05）； compared with the model group， spontaneous pain scores in Tuina group of rats significantly decreased on day 10， 14， and 18 after modelling， and PWTs significantly increased on day 14 and 18 after modelling （P＜0.05）. Compared with Tuina group， spontaneous pain scores increased on day 10， 14， and 18 of modelling， and PWTs decreased at days 14 and 18 of modelling in the MK-801 plus Tuina group had higher spontaneous pain scores on days 10， 14， and 18 after modelling and lower PWTs on days 14 and 18 after modelling （P＜0.05）. Compared with the sham operation group， the neuronal arrangement in the hippocampal CA3 region of the rats in the model group was disordered， with decreased number of Nysted-positive cells and synapses， reduced thickness of postsynaptic densities， length of active bands， and curvature of synaptic interfaces， wider synaptic gaps， and decreased immunofluorescent positive expression of NR2B and PSD95 as well as the expression of immunoblotting proteins in hippocampal CA3 region （P＜0.05）. Compared with the model group， more dense arranged nerve cells， the number of Nysted-positive cells， the number of synapses， the thickness of postsynaptic dense material， the length of active bands increased， the synaptic gap became significantly narrower， and the positive expression of immunofluorescence and immunoblotting protein expression of NR2B， PSD95 increased in the rat hippocampal CA3 region of Tuina group （P＜0.05）. Compared with Tuina group， the neuronal morphology of the hippocampal CA3 region in MK-801 plus Tuina group was severely damaged， and the number of Nystrom's-positive cells， the number of synapses， the thickness of post-synaptic densities， the length of active bands， and the curvature of synaptic interfaces reduced， the synaptic gaps became wider， and the immunofluorescent positive expression of NR2B， PSD95， and the expression of immunostained proteins decreased （P＜0.05）. ConclusionTuina at "Weizhong" （BL 40） showed significant analgesic effect， and one of the possible mechanisms concluded as significantly increasing the levels of NR2B and PSD95 protein expression in hippocampal CA3 region and thus modulating the synaptic plasticity of the hippocampus.