Objective To observe the effect of acupuncture at different time on dorsal root ganglion(DRG)in neuropathic pain(NP)mice.Different regulatory effects of transient receptor potential vanilloid 1(TRPV1)and substance P(SP)in DRG.Methods A total of64 C57BL/6J mice were randomly divided into two intervention groups:ZT8 group and ZT16 group,among which two intervention groups were randomly divided into control group(8 mice),sham-operation group(8 mice),model group(8 mice)and acupuncture group(8 mice).The chronic constriction injury(CCI)model was prepared by sciatic nerve ligation in model group and acupuncture group,and only the sutured wound after sciatic nerve was exposed in sham-operation group.On the 8th day after operation,acupuncture treatment was performed on mice in the acupuncture group.The intervention methods were the same in the two groups,only the acupuncture time was different.The"Yanglingquan"on the surgical side was taken at the acupoint,and manual acupuncture and needle were performed once a day for 15 minutes each time.The mechanical pain and thermal pain thresholds of each group of mice were measured at 3,7,10 and 14 days after modeling to evaluate their pain.The expression of SP and TRPV1 in the L4-6 DRG on the surgical side of mice was observed by immunofluorescence double labeling method,and the SP content in the tissue around the sciatic nerve on the affected side and the skin on the dorsal fibra of the foot was determined by enzymed-linked immunosorbent assay(ELISA).Results ① Acupuncture intervention could improve mechanical and thermal hyperalgesia in CCI mice.And ZT8 time point acupuncture effect is better than ZT16 time point.②Acupuncture can significantly reduce the expression of TRPV1 and SP in DRG of CCI mice,and the effect of acupuncture at ZT8 time point is better than that at ZT16 time point.③Acupuncture can significantly reduce the SP concentration in the peripheral tissue of CCI mice,and the effect of acupuncture at ZT8 time point is better than that at ZT16 time point.④Acupuncture could significantly reduce the SP concentration in the distal skin of the injured nerve in CCI mice,and there was no significant difference between ZT8 and ZT16.Conclusion Acupuncture at different time has different regulation effects on hyperalgesia in CCI mice,and the regulation effect of ZT8 is better than that of ZT16.This may be related to the different regulatory effects of acupuncture at different time on the expression of TRPV1 and SP in DRG of CCI mice and the local SP concentration of injured nerves.

Objective To observe the effect of acupuncture at different time on dorsal root ganglion(DRG)in neuropathic pain(NP)mice.Different regulatory effects of transient receptor potential vanilloid 1(TRPV1)and substance P(SP)in DRG.Methods A total of64 C57BL/6J mice were randomly divided into two intervention groups:ZT8 group and ZT16 group,among which two intervention groups were randomly divided into control group(8 mice),sham-operation group(8 mice),model group(8 mice)and acupuncture group(8 mice).The chronic constriction injury(CCI)model was prepared by sciatic nerve ligation in model group and acupuncture group,and only the sutured wound after sciatic nerve was exposed in sham-operation group.On the 8th day after operation,acupuncture treatment was performed on mice in the acupuncture group.The intervention methods were the same in the two groups,only the acupuncture time was different.The"Yanglingquan"on the surgical side was taken at the acupoint,and manual acupuncture and needle were performed once a day for 15 minutes each time.The mechanical pain and thermal pain thresholds of each group of mice were measured at 3,7,10 and 14 days after modeling to evaluate their pain.The expression of SP and TRPV1 in the L4-6 DRG on the surgical side of mice was observed by immunofluorescence double labeling method,and the SP content in the tissue around the sciatic nerve on the affected side and the skin on the dorsal fibra of the foot was determined by enzymed-linked immunosorbent assay(ELISA).Results ① Acupuncture intervention could improve mechanical and thermal hyperalgesia in CCI mice.And ZT8 time point acupuncture effect is better than ZT16 time point.②Acupuncture can significantly reduce the expression of TRPV1 and SP in DRG of CCI mice,and the effect of acupuncture at ZT8 time point is better than that at ZT16 time point.③Acupuncture can significantly reduce the SP concentration in the peripheral tissue of CCI mice,and the effect of acupuncture at ZT8 time point is better than that at ZT16 time point.④Acupuncture could significantly reduce the SP concentration in the distal skin of the injured nerve in CCI mice,and there was no significant difference between ZT8 and ZT16.Conclusion Acupuncture at different time has different regulation effects on hyperalgesia in CCI mice,and the regulation effect of ZT8 is better than that of ZT16.This may be related to the different regulatory effects of acupuncture at different time on the expression of TRPV1 and SP in DRG of CCI mice and the local SP concentration of injured nerves.

Objective To investigates the effect of acupuncture on sensitization of Zusanli(ST36)in rats with different functional states by using healthy and knee osteoarthritis model rats.Methods Male SD rats were randomly divided into control,model,model-acupuncture and blank-acupuncture group,with 7 rats in each group.KOA rat model was prepared by intra-articular injection of 1 mg·50 μL-1 monoiodoacetic aci(MIA)in model group and model-acupuncture group.On the second day of modeling,acupuncture treatment was performed on the left Zusanli of the model acupuncture group and the blank-acupuncture group,once everyday for 20 min,5 times as a course of treatment,2 days between courses.The general condition,knee joint diameter,plantar thermal pain threshold and Lequesne MG score of rats was observed before modeling and after acupuncture.Observing the morphology of knee joint cartilage to judge whether the model is successful,measuring the mechanical pain threshold of Zusanli to investigate the acupoint sensitization,observing and counting the morphology of skin mast cells in the acupoint area,and detecting the expression of skin calcitonin gene-related peptide(CGRP)in the acupoint area.Results The mechanical pain threshold of Zusanli after acupuncture in model group and blank-acupuncture group decreased significantly after modeling(P<0.01,P<0.05),compared with the control group,the change rate of mechanical pain threshold in model group and blank-acupuncture group increased significantly(P<0.05),compared with the model group,the mechanical pain threshold of Zusanli in the model-acupuncture group decreased significantly(P<0.01).Compared with the control group,the fluorescence intensity of CGRP protein in the skin tissue of Zusanli in the model group increased significantly(P<0.01),MC degranulation rate increased significantly(P<0.05),and there was no significant difference in the fluorescence intensity of CGRP protein of Zusanli in the blank-acupuncture group(P>0.05),MC degranulation rate increased obviously(P<0.01),CGRP protein of Zusanli in the model-acupuncture group was significantly reduced compared with the model group(P<0.01),and there was no significant difference in the degranulation rate of MC(P>0.05).Conclusion Acupoint sensitization can occur in different functional states of rats.Zusanli(ST36)of KOA model rats can be sensitized,and acupuncture stimulation can make Zusanli sensitization caused by disease disappear.Under physiological conditions,acupuncture stimulation can induce similar sensitization phenomenon.