The Study Of Objective Characterization And Morphological Basis Of Needling ST36 De Qi In Rats

Background and Objective:The Yellow Emperor’s Internal Classic says,"The key to the effectiveness of acupuncture is to arrival of qi".When get Qi,it is De Qi.De Qi is considered to be the key to obtain the curative effect of acupuncture,but its underlying mechanism is unclear.The previous research of the research group found that when acupuncture human body arrivals of qi,there are myoelectric signals at the local acupoints,and the needle traction force increases,and both myoelectric signals and needle traction force are positively correlated with the subjective score of De Qi,indicating that these objective indexes can be used to characterize acupuncture De Qi.Further discussion on the scientific basis of De Qi needs to be carried out on the experimental animal platform,taking the manual acupuncture ST36 in the treatment of AA rats as the research platform,the purpose of this study was to explore whether i EMG and needle body traction can characterize acupuncture De Qi in rats.And further study the material basis of De Qi and its relationship with acupuncture effect,so as to provide experimental basis for objectively characterizing the De Qi of experimental animals and revealing the scientific basis of De Qi.MethodResearch content I.Study on objective characterization of needling ST36 De Qi in ratsWistar rats were divided into acupuncture De Qi group and acupuncture without De Qi group.Acupuncture De Qi group: the needle was inserted into the acupoint for 5 ～ 7mm,and the twisting manipulation of mild reinforcing-reducing method was performed(180 times /min,continuous stimulation for 2min,needle retention for 5min,a total of 4 times),the hands of the needle applicator felt heavy and tight.Acupuncture without De Qi group: the needles inserted into the same depth without acupuncture manipulation.The physiological signal acquisition system was used to record the local EMG signals of acupoints during acupuncture and measuring the pulling force of needle body with biosensor.Compare the EMG signal and needle traction force of the two acupuncture operation schemes,and determine whether the rats can produce the objective response of electromyography and needle body pulling force when applying the acupuncture method of De Qi.Research contentⅡ.Study on analgesic effect of acupuncture De Qi in CFA rats and histomorphology of acupoint areaAdult male Wistar rats with stable pain threshold were divided into control group,model group,acupuncture without De Qi group and acupuncture De Qi group,with 6 rats in each group.Except that the bottom of the right foot of the control group was injected with normal saline,the other three groups were injected with the same amount of complete Freund adjuvant(CFA)at the same part to make the AIA pain model.The control group and model group received the same length and degree of restraint measures as the acupuncture group.The acupuncture intervention measures in the acupuncture De Qi group and acupuncture without De Qi group were the same as those used in the first research content.Acupuncture once a day for three days.The effects of acupuncture De Qi and without De Qi manipulation on the plantar pain threshold of rats at different time points were observed by plantar thermal pain tester and plantar tactile instrument.Enzyme linked immunosorbent assay(ELISA)was used to detect the protein contents of cyclooxygenase 2(COX2)and prostaglandin E2(PGE2)after acupuncture intervention.Immunofluorescence staining was used to detect the expression of Phosphorylated N-methyl-D-aspartic receptor(p-NMDAR)closely related to pain signal transduction in the dorsal horn of spinal cord at lumbar distension.Masson staining was used to observe the histomorphological changes of acupoint area.Compare the analgesic effects and differences between the two acupuncture schemes,and explore the morphological basis of acupuncture to De Qi.Research content Ⅲ.Study on the relationship between the material basis of acupuncture ST36 De Qi and acupuncture effect1.Study on the correlation between muscle tissue in acupoints and the analgesic effect of Acupuncture De QiAdult male Wistar rats with stable pain threshold were divided into Control group,AA +vehicle group,AA + SC(succinylcholine chloride solution)group,ACU + vehicle group and ACU + SC group.In the SC group,muscle relaxant was injected into the local muscle of ST36 acupoint to relax the muscle in the acupoint area and temporarily lose the contractile function.In the solvent group,the same amount of muscle relaxant solvent was injected at the same position as the control.The local EMG signals of acupoints during acupuncture were collected to detect the traction force of needle body.The pain threshold of rats was detected by plantar thermal pain tester and plantar tactile instrument.Explore the effects of inhibiting muscle contraction on electromyography,needle traction and analgesic effect of acupuncture De Qi.2.Study on the correlation between collagen fibers in acupoints and the analgesic effect of Acupuncture De QiAdult male Wistar rats with stable pain threshold were divided into Control group,AA +vehicle group,ACU + vehicle group and ACU + collagenase group.Collagenase group injected type I collagenase locally at ST36 acupoint to dissolve collagen fibers in acupoint area.The solvent group was injected with the same amount of collagenase solvent at ST36 as the control.The local EMG signals of acupoints during acupuncture were collected,and the pain threshold of rats was detected by plantar thermal pain tester and plantar tactile instrument to explore the effect of dissolving collagen fibers in acupoint area on EMG and analgesic effect.Result1.EMG signal and needle traction force can be used to characterize acupuncture ST36 De Qi in rats.In the acupuncture stage of acupuncture operation,the EMG release degree of acupuncture De Qi group was significantly greater than that of acupuncture without De Qi group,after converting the obtained EMG data into i EMG value for comparison,it is found that the i EMG value of acupuncture De Qi group is greater than that of acupuncture without De Qi group,with statistical difference.The traction force of the needle body in the acupuncture De Qi group was greater than that in the acupuncture without De Qi group,with statistical difference.2.Acupuncture has a good analgesic effect on adjuvant arthritis rats,and the analgesic effect of acupuncture De Qi manipulation is better.Compared with the control group,it was found that after the adjuvant arthritis model was successfully established,the plantar thermal radiation pain threshold and mechanical pain threshold of rats on the model side decreased,acupuncture can increase the plantar thermal radiation pain threshold and mechanical pain threshold of AIA rats.It shows that acupuncture analgesia is effective,at the same time,the analgesic effect of acupuncture De Qi group appeared earlier than that of acupuncture without De Qi group,and the effect was better.In addition,acupuncture can reduce the expression of p-NMDAR in the spinal dorsal horn at the lumbar distension of the spinal cord,and the reduction effect of acupuncture De Qi group is more obvious.However,the protein contents of COX2 and PGE2,which are pain causing substances in feet,did not decrease significantly.Masson trichrome staining technique was used to observe the local tissue morphology of acupuncture points.It showed that the needle path of acupuncture De Qi operation was obviously distributed in the muscle layer,accompanied by many collagens fiber hyperplasia.It shows that the muscle tissue and collagen fibers in the local acupoints may be the material basis of acupuncture De Qi.3.As the morphological basis of acupuncture De Qi,the muscle tissue and collagen fiber in acupoint area mediate the analgesic effect of acupuncture De Qi,in which the function of muscle tissue is more important than that of collagen fiber.After local intramuscular injection of succinylcholine chloride solution at ST36 in rats,the degree of EMG induced by acupuncture De Di decreased significantly,and it can completely antagonize the analgesic effect of acupuncture De Qi.After local intramuscular injection of type I collagenase at ST36 in rats,Masson staining technique observed that the arrangement of collagen fibers in subcutaneous fascia layer and muscle layer in the point area was loose and the staining became lighter,which proved that the effect of dissolving collagen fibers in the point area was achieved.Injection of collagenase into acupoints could partially antagonize the analgesic effect of acupuncture De Qi,but had no significant effect on the EMG signal induced by acupuncture De Qi.Conclusion1.The degree of EMG release in the rat acupoint area and the traction force on the body when the needle is out can objectively characterize the acupuncture ST36 De Qi of rats,which is consistent with the objective characterization index of acupuncture De Qi in human experiment.2.Acupuncture at ST36 can effectively relieve the pain symptoms of adjuvant arthritis in rats,and the analgesic effect of acupuncture De Qi manipulation is better.Muscle tissue and collagen fibers in acupoint area may be the morphological basis of acupuncture De Qi.3.Both muscle tissue and collagen fibers mediate the analgesic effect of acupuncture De Qi,and the effect of muscle tissue is more important than that of collagen fibers.