| Purpose: In this study,Mycoscia pain syndrome(MPS)model rats were replicated by local shock combined with centrifugal motion,and Myofascial Trigger Points(MPS)were observed.Mtrps)was used as the therapeutic point.The therapeutic effects of acupuncture and moxibustion on MPS rats were observed.By observing the effects of acupuncture and moxibustion pain point on local tissue morphology,pro-inflammatory factors,pain factors and the expression of pain transmitters related to spinal dorsal horn,the therapeutic effects and mechanism of acupuncture and moxibustion different ways of intervening pain point on MPS rats were explored,so as to provide experimental basis for acupuncture and moxibustion treatment of MPS.Materials and Methods: 1.Grouping: 32 male SD rats(SPF grade,weight 220-250g)were divided into control group,model group,acupuncture group and moxibustion group,with 8 rats in each group 2.Modeling: rats in model group,acupuncture group and moxibustion group were used to make MTr Ps model by striking combined with centrifugal movement.The specific steps were as follows: on the first day of each experimental cycle,according to the weight of rats,10% chloral hydrate(3ml / kg)was injected into the abdomen of rats for anesthesia,and the left lower limb medial thigh muscle of rats was struck once by self-made striking device;on the second day,10% chloral hydrate(3ml / kg)was injected into the abdomen of rats,The rats were asked to run downhill at the speed of 16 m / min on the electric track with a gradient of-16 ° for 90min;the rats were fed normally for the other 3 to 7 days without intervention.The above process lasted for 8 weeks,which was the expectation of intervention.After 4 weeks,the rats were fed normally without any intervention,which was the recovery period 3.Interventions:(1)acupuncture group: 0.35 mm × 40 mm filiform needle was used to prick through the nodule of pain stimulation point.After the local convulsive reaction was stimulated by lifting,inserting and twisting,the electroacupuncture instrument was connected and adjusted to the density wave with frequency of 2-20 hz.The appropriate intensity was slight tremor on the left lower limb of rats,lasting for 15 min,once a day for 1 week.(2)moxibustion group: the moxa stick was placed 2-3cm above the pain stimulation point,Control the skin temperature at(46 ±1)? and apply moxibustion for 15 min to ensure that there is no swelling,scald,blood,tissue fluid exudation and other infection at the moxibustion site.Treat once a day for one week.(3)Control group,model group: in the same environment to take the same way to capture and control,and do not take any intervention measures 4.Sampling: in the morning of the next day after the treatment,the rats were anesthetized with 10% chloral hydrate(3ml / kg)by abdominal injection,and then the rats were fixed on the fixed plate in the supine position to straighten the left lower limb,The samples were cut off at the attachment of skeletal muscle and immersed in 4% paraformaldehyde solution for 24 hours for pathological detection.Abdominal aorta blood was taken from abdominal aorta at 3000 R / min,centrifuged at 4 ? for 10 minutes,and the supernatant was separated and placed in-80 ? ultra-low temperature refrigerator for subsequent enzyme-linked immunosorbent assay(ELISA)detection;L4-6 lumbar enlargement of spinal cord was taken after blood collection,Some of them were immersed in 4% paraformaldehyde fixed solution and stored for immunohistochemical detection,and some were used for real-time reverse transcription polymerase chain reaction(RT-PCR)detection 5.Detection: behavior and hot plate test were used to detect the latent period of rat's hot feet contraction;electromyography was used to detect the spontaneous potential and activity frequency of left lower limb medial femoral muscle;pathomorphological method was used to observe the morphological changes of muscle fibers in left lower limb medial femoral muscle;ELISA was used to detect the concentration of prostaglandin E2(PGE2)in serum of rats;Immunohistochemical method was used to detect the quantitative analysis of immediate early gene c-fos(IEG)in spinal dorsal horn of rats,and real-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR)was used to detect the gene expression of neuronal nitric oxide synthase(n NOS)in spinal cord.Results: 1 Changes in the incubation period of left lower extremities of rats After modeling,no abnormal gait,licking and spontaneous leg lifting were observed in the control group,and the incubation period of hot foot contraction was not significantly changed compared with that before modeling.Compared with the control group,the model group,the acupuncture group and the moxibustion group all had lameness,and the latency of heat shrinkage was significantly decreased(P<0.05).Compared with the model group,there were no significant differences in the incubation period and gait of the acupuncture group and moxibustion group(P>0.05).After treatment,compared with the control group,lameness could be seen occasionally in the model group,and the gait of the rats in the acupuncture and moxibustion groups was basically normal,but the latent time of thermal foot withdrawal in the left lower extremity of the rats in the three groups was significantly decreased(P<0.05).Compared with the model group,the incubation period of heat shrinkage of left lower limb in acupuncture group and moxibustion group was significantly improved,with statistical significance(P<0.05).2.Changes in spontaneous potential and activity frequency of the rats in each group the em G results showed that: In the resting state,the em G of the control group was like a straight line without abnormal spontaneous potential,indicating that there was no MTRP at the vastus medialis of the left lower limb of the rats;Compared with the control group,a series of background potentials with high frequency and low amplitude and peak potentials with relatively high amplitude appeared in the model group,lasting for 0.5-10 min.The frequency of spontaneous potential activity changed significantly,with statistically significant difference(P < 0.05).Compared with the model group,occasional spontaneous electrical activity was observed in both the MTRP group and the moxibustion group,and the frequency decreased significantly,the difference was statistically significant(P < 0.05).3.Morphological changes of left lower limb medial myopathy of rats in the control group: In the control group,most of the muscle cells in the cross section of the affected muscle presented regular and orderly shape,which could clearly distinguish the fiber structure.The cells were uniform in size,arranged closely,orderly and regularly,and multiple nuclei could be seen at the edge of the cells,and the nuclei were located under the sarcolemma,and there was no inlfammatory nuclear migration and inflammatory infiltration.In the longitudinal section,the muscle fibers were arranged closely and regularly,with uniform thickness,Most of the nuclei were distributed under the membrane of muscle fibers.In the model group,the size and morphology of muscle cells were changed,the cells were arranged disorderly,the inflammatory nuclei moved inward and infiltrated,and a large area of tissue adhesion appeared at the contracture node.In the longitudinal section,the muscle fibers were different in thickness,arranged disorderly,and a large area of adhesion area appeared at the contracture node,In the acupuncture group and moxibustion group,the shape of muscle cells was almost normal in the transverse section,and round or oval muscle cells were occasionally seen in the acupuncture group,and the phenomenon of nuclear inward shift was more obvious than that in the acupuncture group;in the longitudinal section,the muscle fibers in the acupuncture group and moxibustion group gradually recovered,and slight atrophy,degeneration and inflammatory cell infiltration were seen,And the muscle tissue morphology of moxibustion group was closer to that of control group 4.Compared with the control group,the expression of PGE2 protein in the serum of the model group,Compared with the control group,serum PGE2 concentrations in model group were increased,with statistical significance(P<0.05).Compared with model group,serum PGE2 concentrations in acupuncture group and moxibustion group were significantly decreased,with statistical significance(P<0.05).Compared with the moxibustion group,the concentration of PGE2 in the acupuncture group was increased,and the difference was statistically significant(P<0.05).5.Changes in the expression of quantitative optical density of C-fos immunohistochemical integral in spinal dorsal horn of rats in each group In the control group,C-fos positive neurons were seldom expressed in the dorsal horn of the spinal cord.Most of them showed blue negative cells,and the number was small.Compared with the control group,the expression of C-fos positive neurons in the model group,the moxibustion group and the acupuncture group was round or oval,and the nuclei were brown and yellow.The higher integral quantitative optical density value was significantly different from that in the control group,with statistical significance(P<0.05).Compared with the model group,the C-FOS staining of the acupuncture group and the moxibustion group was slightly lighter,and the difference in the reduction of optical density was statistically significant(P<0.05).Moreover,the optical density of the acupuncture group and the moxibustion group was lower,and the difference was statistically significant(P<0.05).6 Changes of n NOS gene expression in spinal cord tissue of rats in each group Compared with the control group,the expression of n NOS gene in spinal cord tissue of model group,moxibustion group and acupuncture group were significantly increased(P<0.05);Compared with model group,the expression of n NOS gene in spinal cord tissue of acupuncture group and moxibustion group was lower than that in the control group(P<0.05);Compared with acupuncture group,the expression of n NOS gene in moxibustion group was significantly increased,and the acupuncture treatment effect was better than moxibustion(P<0.05).Conclusion: The stimulation pain point is the core of diagnosis and treatment of MPS.Acupuncture and moxibustion can increase the thermal pain threshold of MPS rats,reduce the abnormal spontaneous potential of the local muscles where the stimulation pain point is and restore the muscle cell morphology,thus inactivation pain point to effectively improve chronic myofascial pain.Acupuncture and moxibustion can inhibit the synthesis and release of PGE2,reduce the inflammatory reaction of MPS,promote local tissue recovery and directly reduce the sensitivity of peripheral receptors The signal intensity of nociceptive stimulation was weakened,the synthesis and release of NNOS and C-fos related neurotransmitters were interfered,and the activation of pain transmitters in spinal Dorsal Horn was inhibited.Based on the difference of local therapeutic mechanism between acupuncture and moxibustion,acupuncture stimulation pain spot is better than moxibustion in regulating the expression of n NOS and C-fos in Dorsal Horn of spinal cord The down-regulation of PGE2 expression and the restoration of Muscle cell morphology by moxibustion stimulation were better than that by acupuncture stimulation,which indicated that moxibustion therapy was better at local tissue repair,peripheral sensitization and inflammatory environment intervention. |
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