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 observe the effects of tuina on P2Y12/RhoA/ROCK2 pathway and c-Fos protein expression of microglia in spinal dorsal horn of sciatic nerve chronic compressive injury(CCI)rats;To explore the mechanism of tuina in treating lumbar disc herniation.Methods A CCI model of right sciatic nerve was used to simulate neuropathic pain in lumbar disc herniation.24 male SD rats were randomly divided into blank group,model group and tuina group,with 8 rats in each group.After 4 days of modeling,massage intervention was used to in the tuina group for 14 days.The paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)of rats before and on the 4th,10th,and 17th day after modeling were observed,immunofluorescence staining was used to detect the expression of Iba1 and P2Y12 protein in right spinal dorsal horn,Western blot was used to detect the expressions of RhoA and ROCK2 protein in right spinal dorsal horn,immunohistochemistry staining was used to detect the number of c-Fos positive cells in right spinal dorsal horn.Results Compared with the blank group,the PWT and PWL of the model group were significantly reduced on the 4th,10th,and 17th day after modeling(P<0.001),while the expressions of Iba1,P2Y12,RhoA and ROCK2 protein in right spinal dorsal horn significantly increased(P<0.001,P<0.05),the number of c-Fos positive cells significantly increased(P<0.001).Compared with the model group,the PWT and PWL of the tuina group significantly increased on the 10th and 17th day after modeling(P<0.05,P<0.01,P<0.001),and the expressions of Iba1,P2Y12,RhoA and ROCK2 protein in right spinal dorsal horn significantly decreased(P<0.001,P<0.01,P<0.05),and the number of c-Fos positive cells significantly reduced(P<0.001).Conclusion Tuina may inhibit the activation of microglia by regulating P2Y12/RhoA/ROCK2 pathway and c-Fos protein expression in spinal dorsal horn,reduce neuronal excitability,and exert analgesic effects on lumbar disc herniation.

Background:Previous study has found that harmine inhibited the proliferation of gastric cancer cells by down-regulating cyclooxygenase-2(COX-2)expression. However,its molecular mechanism is not fully clear. Aims:To investigate the effect of harmine on proliferation and apoptosis of gastric cancer cells,and explore the role of PTEN/Akt/MDM2 signaling pathway in this process. Methods:Human gastric adenocarcinoma cell line SGC-7901 and MKN-45 were treated with harmine at different concentrations(2,4,8,16,32 μg/mL)for 24,48,and 72 hours. The cell proliferation and apoptosis were detected by MTT assay and Hoechst staining,respectively. The expressions of PTEN,COX-2, phosphorylated Akt(p-Akt)and p-MDM2 were measured by Western blotting. Results:Harmine dose- and time-dependently inhibited proliferation and induced apoptosis of SGC-7901 and MKN-45 cells. Also,harmine dose-dependently increased PTEN expression,and inhibited p-Akt,p-MDM2 and COX-2 expressions in SGC-7901 and MKN-45 cells. Conclusions:Harmine may inhibit proliferation and induce apoptosis of gastric cancer cells via down-regulating COX-2 expression through PTEN/Akt/MDM2 signaling pathway.