ObjectiveTo investigate the effect of icariin （ICA） on the phenotype of dendritic cells （DCs） in heart tissue of the Dahl salt-sensitive myocardial remodeling model of rats and its regulation on the vitamin D system. MethodsMale Dahl salt-resistant rats were divided into a normal group， and male Dahl salt-sensitive rats were divided into a model group， low-， medium-， and high-dose ICA groups （30， 60， 120 mg·kg^-1·d^-1）， and Vitamin D group （3×10^-5 mg·kg^-1·d^-1）. In addition to the normal group， the other groups were given an 8% high salt diet to establish a myocardial remodeling model and received intragastric administration after successful modelling once a day for six weeks. The dynamic changes in tail artery blood pressure were monitored， and detection of cardiac ultrasound function in rats was performed. Hematoxylin-eosin （HE） staining and Masson staining were used to observe the morphological changes in rat heart tissue. The phenotype of DCs and T helper cell 17 （Th17）/regulatory T cell （Treg） ratio were detected by flow cytometry. The mRNA and protein expression of vitamin D receptor （VDR）， 1α-hydroxylase （CYP27B1）， 24-hydroxylase （CYP24A1）， forkhead frame protein 3 （FoxP3）， solitaire receptor γt （RORγt）， myocardial type Ⅰ collagen （ColⅠ）， and type collagen （ColⅢ） in heart tissue was detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultsCompared with the normal group， the model group showed disordered arrangement and rupture of myocardial cells， nuclear condensation， significant edema of myocardial tissue， significant proliferation of collagen fibers in a network distribution， and a significant increase in tail artery blood pressure， left ventricular end diastolic diameter （LVEDD）， and left ventricular end systolic diameter （LVESD） （P<0.05）. The phenotype of cardiac DCs was CD40， CD80， and CD86， and the levels of major histocompatibility complex Ⅱ （MHC-Ⅱ）， Th17 cells， and Th17/Treg were significantly increased （P<0.05）. The mRNA and protein expression of CYP24A1 and RORγt in the heart， as well as the mRNA expression of ColⅠ and ColⅢ， were significantly increased （P<0.05）. The left ventricular ejection fraction （LVEF）， interventricular septal thickness （IVSD）， and left ventricular posterior wall thickness （LVPWD） were significantly decreased （P<0.05）. The phenotype of cardiac DCs such as CD11， CD11b， and Treg cells， were significantly reduced （P<0.05）， while the mRNA and protein expression of cardiac VDR， CYP27B1， and FoxP3 were significantly decreased （P<0.05）. Compared with the model group， the low-， medium-， and high-dose ICA groups and vitamin D group significantly reduced myocardial cell rupture and nuclear consolidation in rats. The high-dose ICA group and vitamin D group showed a small amount of myocardial cell rupture and nuclear consolidation， improving myocardial fiber arrangement to varying degrees and significantly reducing myocardial fiber rupture and proliferation. The tail artery blood pressure， LVEDD， and LVESD were significantly decreased in the low-， medium-， and high-dose ICA groups and vitamin D group （P<0.05）， and the phenotype of cardiac DCs including CD40， CD80， CD86， MHC-Ⅱ， Th17 cells， and Th17/Treg were significantly decreased （P<0.05）. The mRNA and protein expression of CYP24A1 and RORγt， and the mRNA expression of ColⅠ and ColⅢ in the heart were significantly decreased in the medium- and high-dose ICA groups and vitamin D group （P<0.05）. The LVEF， IVSD， and LVPWD of myocardial remodeling model rats in the low-， medium-， and high-dose ICA groups and vitamin D group were significantly increased （P<0.05）. The phenotypes of cardiac DCs including CD11， CD11b， and Treg cells were significantly increased in the medium- and high-dose ICA groups and the Vitamin D group （P<0.05）. The mRNA and protein expressions of VDR， CYP27B1， and FoxP3 in the heart were significantly increased in the medium- and high-dose ICA groups and vitamin D group （P<0.05）. ConclusionICA can regulate tail artery blood pressure， cardiac structural and functional damage， and myocardial tissue fibrosis and inhibit phenotype and functional maturation of DCs in heart tissue in the myocardial remodeling model of Dahl salt-sensitive rats. It can also affect the gene and protein expression of VDR， CYP24A1， and CYP27B1， achieving its intervention in Th17/Treg balance in the immune process of myocardial remodeling possibly by regulating vitamin D/VDR in heart tissue.

Objective To investigate the effects and potential mechanism of tuina on pain and depressive behaviors in rats with neuropathic pain(NP).Methods A total of 102 SD rats were randomly divided into blank group,sham-operation group,model group,tuina group,inhibitor group and inhibitor+tuina group,with 17 rats in each group.The NP model was established by chronic constriction injury of the sciatic nerve.Starting from the 8th day post-operation,the rats underwent a 14-day tuina intervention and stereotactic injection of the SIRT1 inhibitor EX-527(20 μg/μL,0.5 μL)into the hippocampal CA1 region.Pain behaviors were assessed using the mechanical withdrawal threshold test one day before operation and on days 7,14 and 21 post-operation.Depressive behaviors were evaluated using the forced swimming test and sucrose preference test.Nissl staining was employed to observe neuronal morphology and quantity in the hippocampal tissue,while Golgi staining was used to examine dendritic spine density,hippocampal expression of SIRT1/BDNF/TrkB signaling pathway related protein and mRNA were analyzed using immunofluorescence,Western blot and RT-qPCR.Results Compared with the sham-operation group,the model group showed a significant decrease in mechanical withdrawal threshold(P<0.001),prolonged immobility time in the forced swimming test and reduced sucrose preference(P<0.001)on days 7,14 and 21 post-operation;the morphology of hippocampal CA1 neurons was abnormal,with a significant decrease in the number of Nissl positive cells(P<0.001)and a significant decrease in dendritic spine density(P<0.001);the expressions of SIRT1,BDNF and TrkB in dentate gyrus of the hippocampus were significantly reduced(P<0.01,P<0.001),and the protein and mRNA expressions of SIRT1,BDNF and TrkB were significantly reduced(P<0.001).Compared with the model group,the tuina group showed a significant increase in mechanical withdrawal threshold(P<0.01,P<0.001)on days 14 and 21 post-operation,shortened immobility time in the forced swimming test(P<0.01,P<0.001)and increased sucrose preference(P<0.001);the hippocampal CA1 neuronal morphology was improved,with significantly increased Nissl positive cells(P<0.001)and dendritic spine density(P<0.001);the expressions of SIRT1,BDNF and TrkB in dentate gyrus of the hippocampus significantly increased(P<0.01,P<0.001),and the protein and mRNA expressions of SIRT1,BDNF and TrkB were significantly increased(P<0.001).The beneficial effects of tuina were significantly inhibited when the SIRT1 inhibitor EX-527 was used.Conclusion Tuina may alleviate pain and depressive behaviors in NP rats by activating the SIRT1/BDNF/TrkB signaling pathway and improving hippocampal neuronal structural plasticity.

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 postsynaptic dense zone decreased,and the synaptic cleft decreased,while the protein expressions of NR2A,SNAP25 and VGLUT2 in the spinal dorsal horn decreased(P<0.05,P<0.001),and the content of glutamate decreased(P<0.01).Conclusion Tuina may regulate the content of glutamate through the SNAP25/VGLUT2 pathway in the spinal dorsal horn,improve the synaptic ultrastructure of neurons,and have an analgesic effect on lumbar disc herniation.

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 postsynaptic dense zone decreased,and the synaptic cleft decreased,while the protein expressions of NR2A,SNAP25 and VGLUT2 in the spinal dorsal horn decreased(P<0.05,P<0.001),and the content of glutamate decreased(P<0.01).Conclusion Tuina may regulate the content of glutamate through the SNAP25/VGLUT2 pathway in the spinal dorsal horn,improve the synaptic ultrastructure of neurons,and have an analgesic effect on lumbar disc herniation.

Objective To investigate the effects and potential mechanism of tuina on pain and depressive behaviors in rats with neuropathic pain(NP).Methods A total of 102 SD rats were randomly divided into blank group,sham-operation group,model group,tuina group,inhibitor group and inhibitor+tuina group,with 17 rats in each group.The NP model was established by chronic constriction injury of the sciatic nerve.Starting from the 8th day post-operation,the rats underwent a 14-day tuina intervention and stereotactic injection of the SIRT1 inhibitor EX-527(20 μg/μL,0.5 μL)into the hippocampal CA1 region.Pain behaviors were assessed using the mechanical withdrawal threshold test one day before operation and on days 7,14 and 21 post-operation.Depressive behaviors were evaluated using the forced swimming test and sucrose preference test.Nissl staining was employed to observe neuronal morphology and quantity in the hippocampal tissue,while Golgi staining was used to examine dendritic spine density,hippocampal expression of SIRT1/BDNF/TrkB signaling pathway related protein and mRNA were analyzed using immunofluorescence,Western blot and RT-qPCR.Results Compared with the sham-operation group,the model group showed a significant decrease in mechanical withdrawal threshold(P<0.001),prolonged immobility time in the forced swimming test and reduced sucrose preference(P<0.001)on days 7,14 and 21 post-operation;the morphology of hippocampal CA1 neurons was abnormal,with a significant decrease in the number of Nissl positive cells(P<0.001)and a significant decrease in dendritic spine density(P<0.001);the expressions of SIRT1,BDNF and TrkB in dentate gyrus of the hippocampus were significantly reduced(P<0.01,P<0.001),and the protein and mRNA expressions of SIRT1,BDNF and TrkB were significantly reduced(P<0.001).Compared with the model group,the tuina group showed a significant increase in mechanical withdrawal threshold(P<0.01,P<0.001)on days 14 and 21 post-operation,shortened immobility time in the forced swimming test(P<0.01,P<0.001)and increased sucrose preference(P<0.001);the hippocampal CA1 neuronal morphology was improved,with significantly increased Nissl positive cells(P<0.001)and dendritic spine density(P<0.001);the expressions of SIRT1,BDNF and TrkB in dentate gyrus of the hippocampus significantly increased(P<0.01,P<0.001),and the protein and mRNA expressions of SIRT1,BDNF and TrkB were significantly increased(P<0.001).The beneficial effects of tuina were significantly inhibited when the SIRT1 inhibitor EX-527 was used.Conclusion Tuina may alleviate pain and depressive behaviors in NP rats by activating the SIRT1/BDNF/TrkB signaling pathway and improving hippocampal neuronal structural plasticity.

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.

Objective To explore the effect of chemotherapeutic drugs doxorubicin or cisplatin on lipid metabolism of macrophages and its regulatory mechanism.Methods Macrophage RAW264.7 was treated with doxorubicin or cisplatin,and intracellular lipid level was detected by oil red O and ELISA;RNA sequence screen-ing and Western blot were used to confirm the changes of gene expression after chemotherapeutic drug treatment;The effects of silencing ROBO3 on cellular lipid metabolism were explored,and changes in key target genes of lipid metabolism were detected by Q-PCR and western blot.Results Adriamycin or cisplatin induced disturbances in macrophage cholesterol metabolism and exacerbated macrophage foaminess.Further studies showed that the expression of the axon guidance factor receptor,ROBO3,increased and then decreased during the chemotherapeutic drug-induced macrophage foaming process.Further intervention with ROBO3 exacerbates oxldl-induced cholesterol accumulation and foam formation in macrophages.Mechanistically,ROBO3 deficiency promotes the expression of cholesterol synthesis-related gene DHCR24 and inhibits the expression of cholesterol elimination-related gene ABCG1,resulting in cholesterol accumulation in macrophages.Conclusion This study found that ROBO3 plays an important regulatory role in the disruption of cholesterol metabolism and its foaming process in macrophages induced by chemotherapeutic drugs,which may provide new targets and ideas for the prevention and treatment of chemotherapy-related atherosclerosis.

ObjectiveTo investigate the effect of icariin （ICA）-mediated vitamin D system on peripheral blood dendritic cells （DCs） and helper T cells 17 （Th17）/regulatory T cells （Treg） balance in myocardial remodeling model of Dahl salt-sensitive rats. MethodFifty SPF Dahl salt-sensitive rats were divided into model group， vitamin D group （3×10^-5 mg·kg^-1·d^-1）， and high-， medium-， and low-dose ICA groups （120， 60， 30 mg·kg^-1·d^-1）， and 10 Dahl salt-resistant rats were used as normal group. The myocardial remodeling model was established by feeding rats with a high-salt diet containing 8% NaCl. After six weeks of modeling， the normal group and the model group were given an equal volume of ultrapure water by gavage， and other groups were continuously administrated for six weeks. Cardiac echocardiography， hematoxylin-eosin （HE） staining， and Masson staining were used to observe the pathological changes in cardiac structure and fibrosis. The levels of serum 25（OH）D₃， B-type N-terminal pro-brain natriuretic peptide （NT-ProBNP）， interleukin （IL）-17， transforming growth factor （TGF）-β₁， IL-12， and IL-10 were detected by enzyme-linked immunosorbent assay （ELISA）. The phenotype of peripheral blood DCs and the ratio of Th17/Treg cells of rats were detected by flow cytometry. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the mRNA and protein expressions of vitamin D receptor （VDR），1α-hydroxylase （CYP27B1）， and 24-hydroxylase （CYP24A1） in peripheral blood DCs of rats. ResultCompared with the control group， the rats in the model group had pathological changes such as disordered arrangement of myocardial cells and cytoplasmic hypertrophy and swelling. Myocardial collagen fibers proliferated significantly， and the arrangement of myocardial fibers was disordered. The levels of serum 25（OH）D₃ and IL-10 were significantly decreased， and the levels of serum IL-17， TGF-β₁， IL-6， IL-12， and NT-ProBNP were significantly increased （P<0.05）. The costimulatory molecules CD40， CD80， CD86， and MHC-Ⅱ were highly expressed in the peripheral blood DCs， and the expression of CD11 and CD11b was lower （P<0.05）. The proportion of Th17 cells in the peripheral blood was significantly increased， and the proportion of Treg cells was decreased. The ratio of Th17/Treg was increased （P<0.05）. The mRNA and protein expressions of CYP24A1 in peripheral blood DCs increased， and the mRNA and protein expressions of CYP27B1 and VDR decreased （P<0.05）. Compared with the model group， the arrangement of myocardial fibers in each drug administration group was relatively regular， and the swelling of myocardial cells was significantly reduced. The pathological morphology of myocardial tissue was improved to varying degrees. The pathological changes in myocardial tissue were improved and alleviated to varying degrees. The drug could reduce the serum levels of NT-ProBNP， IL-17， TGF-β₁， IL-6， and IL-12 and increase the level of serum 25（OH）D₃ and IL-10 （P<0.05）. The expression of costimulatory molecules CD40， CD80， CD86， and MHC-Ⅱ in the peripheral blood DCs of rats was decreased， and the expression of CD11 and CD11b molecules was increased （P<0.05）. The drug could reduce the proportion of Th17 cells in peripheral blood and the ratio of Th17/Treg cells and increase the proportion of Treg cells （P<0.05）. It could decrease the mRNA and protein expressions of CYP24A1 in peripheral blood DCs of rats and elevate the mRNA and protein expression of VDR and CYP27B1 （P<0.05）. ConclusionICA can regulate the phenotype of peripheral blood DCs and the ratio of Th17/Treg cells by regulating the vitamin D system and play a role in improving myocardial remodeling from the perspective of immune balance.

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.

Objective To observe the effect of tuina on glutamate uptake and synaptic cleft in the spinal dorsal horn of rats with neuropathic pain through astrocyte NDRG2/GLT-1 pathway,and to explore the potential analgesic mechanism of tuina on neuropathic pain.Methods A total of 54 SD rats were randomly divided into naive group,model group and tuina group(n=18).The CCI model was established in the model group,and the tuina group was treated with acupressure at"Weizhong"(BL 40)from the 4th day after CCI model was successfully established for 14 days.The changes of paw withdrawal threshold at different time points were observed to evaluate the analgesic effect of tuina.Immunofluorescence was used to observe the co-expression of NDRG2?GLT-1 and astrocytes in the spinal dorsal horn.The mRNA levels of NDRG2 and GLT-1 in astrocytes were detected by quantitative real time polymerase chain reaction.The concentrations of glutamate in synaptic cleft were measured by liquid chromatography coupled to tandem mass spectrometry.The width of the synaptic cleft was observed by transmission electron microscopy.Results Compared with the naive group,the paw withdrawal threshold in the CCI group decreased continuously(P<0.01).The number of NDRG2 and GFAP co-labeling positive cells in the spinal dorsal horn increased significantly(P<0.01),and the number of GLT-1 and GFAP co-labeled positive cells was significantly reduced(P<0.01).NDRG2 mRNA expression was up-regulated and GLT-1 mRNA expression decreased in astrocytes(P<0.01).The concentrations of glutamate increased significantly(P<0.01);The synaptic cleft was significantly narrowed(P<0.05).After tuina intervention,the above trend was significantly reversed.Compared with the model group,the paw withdrawal threshold of the tuina group increased from 11 days after CCI(P<0.01),the number of NDRG2 and GFAP co-labeling positive cells in the spinal dorsal horn was significantly reduced(P<0.01),and the number of GLT-1 and GFAP co-labeled positive cells increased significantly(P<0.01);down-regulated the expression of NDRG2 mRNA and restored GLT-1 mRNA expression in astrocytes(P<0.01);decreased glutamate concentration in synaptic cleft(P<0.05),the synaptic cleft was relatively widened(P<0.05).Conclusion Tuina alleviates pain in CCI rats at the spinal cord level possibly by promoting the uptake of glutamate by NDRG2/GLT-1 pathway in astrocytes,restoring the width of synaptic cleft and reversing synaptic plasticity.