| Objective To observe the effects of electroacupuncture on neurotransmitters SP,5-HT,mast cells,astrocytes and inflammatory factors in the dorsal horn of the lumbar spinal cord in fibromyalgia model rats,and to explore the effect and mechanism of electroacupuncture on fibromyalgia.Methods36 male SD rats(body weight 150±20g)were selected.Twelve rats were randomly selected as blank control group(hereinafter referred to as blank group).Another 24 rats were modeled under isoflurane(1%)anesthesia by injecting 2-morpholinol ethanolsulfonic acid along the left tibialis anterior muscle of rats on days 0 and 5.On the 0th and 14 th days,the behavioral changes of the mechanical withdrawal reflex threshold of the rats were observed to confirm the successful modeling.On the 15 th day,the successful modeling rats were randomly divided into the model control group(hereinafter referred to as the model group).Electroacupuncture treatment group(hereinafter referred to as electroacupuncture group),each group with 12 rats.Electroacupuncture group: On the 15 th day after the first injection of 2-morpholinol ethanolsulfonic acid solution,the FM model rats were bound in a fixed frame,and the needles were inserted straight into Zusanli and Yangling quan in the left hind limb for about 4 days.mm,connected to a Korean-style electro-acupuncture instrument,the parameters were 1 m A,2 Hz,the treatment time was 20 min,once every 2 days,and a total of 7 interventions were performed.Model group and blank group: they were only tied to the fixed frame,and the intervention course was the same as that of electroacupuncture.The mechanical paw withdrawal pain threshold of the rats in each group was detected by behavior;the morphological changes of the tibialis anterior muscle of the rats in each group were observed by HE staining;The expression levels of GFAP and 5-HT;the expression of TNF-α,IL-1β,IL-6 and IL-10 in the lumbar spinal cord of rats was detected by suspension chip method.Results1.Results of mechanical withdrawal pain threshold: There was no difference in the mechanical withdrawal pain threshold of rats in each group before modeling;after modeling,compared with the blank group,the mechanical withdrawal pain threshold of rats in the model group was significantly reduced(P < 0.05).after intervention,compared with the model group,the pain threshold of mechanical withdrawal in the electroacupuncture group was significantly increased(P < 0.05).2.HE staining results(1)Transverse section of tibialis anterior muscleBlank group: The muscle fibers of the tibialis anterior muscle are polygonal,the muscle fibers are neatly arranged,the structure of the outer,intramuscular,and muscle bundles is clear,and the sarcoplasm is rich.The inner surface of each muscle fiber has multiple closely attached nuclei,and the size and coloring of the nuclei are uniform,no abnormal changes were found.model group: muscle fibers were disordered and irregular,thickened and edema,there were gaps in muscle fibers,muscle cells were destroyed,gaps between muscle fibers were enlarged,and cell nuclei were transferred to muscle fibers for inflammatory infiltration.electroacupuncture group: muscle fiber destruction decreased,cells The damage was swollen,the arrangement was still neat,and the inflammatory infiltration was not obvious,which was lighter than that of the model group.(2)Coronal plane of tibialis anterior muscleBlank group: muscle fibers were rectangular,neatly arranged,with obvious light and dark bands,neatly arranged nuclei,and uniform coloration;model group: muscle fibers ruptured,with obvious inflammatory infiltration.electroacupuncture group: no rupture of muscle fibers,reduced inflammatory infiltration.3.Immunofluorescence histochemistry results(1)Effects of electroacupuncture on SP content in the spinal dorsal horn of FM model rats SP nerve fibers were mainly detected in layer II of the dorsal horn of the spinal cord.Compared with the blank group,the dorsal horn of the spinal cord in the model group SP significantly increased(P<0.05).compared with the model group,SP in the dorsal horn of the spinal cord in the electroacupuncture group was significantly decreased(P<0.05).(2)Effects of electroacupuncture on the content of MC and 5-HT in the dorsal horn of spinal cord in fibromyalgia model ratsCompared with the blank group,the shape of the MC in the dorsal horn of the spinal cord became smaller in the model group after degranulation,but the number did not change significantly(P>0.05).compared with the model group,the number of MCs in the dorsal horn of the spinal cord in the electroacupuncture group did not change significantly(P>0.05).Compared with the blank group,the amount of 5-HT in the dorsal horn of the spinal cord in the model group was significantly decreased(P<0.05).compared with the model group,the amount of 5-HT in the electroacupuncture group was significantly increased(P<0.05).(3)The effect of electroacupuncture on the content of GFAP in the spinal dorsal horn of fibromyalgia model ratsCompared with the blank group,the GFAP cell body in the dorsal horn of the spinal cord in the model group became larger,the protrusions increased,and the expression increased significantly(P<0.05).Compared with the model group,the GFAP cell body in the dorsal horn of the spinal cord in the electroacupuncture group became smaller,the number of protrusions decreased.The expression was significantly decreased(P<0.05).4.Results of suspension chip methodCompared with the control group,the positive expressions of TNF-α,IL-1β and IL-6 in the model group were significantly increased,and the positive expressions of IL-10 were significantly decreased(P<0.05).The positive expressions of-1β and IL-6 were significantly decreased,and the positive expressions of IL-10 were significantly increased(P<0.05).Conclusions1.Electroacupuncture can improve the pain threshold of mechanical withdrawal in FM model rats.2.Electroacupuncture can promote the repair of tibialis anterior muscle injury in FM model rats.3.Electroacupuncture reduces the expression of the neurotransmitter SP-positive nerve fibers in the superficial dorsal horn of the spinal cord,reduces the expression of activated MC proinflammatory factors and GFAP,and increases the expression of the neurotransmitter 5-HT.The possible mechanism of the analgesic effect in mice.4.Electroacupuncture reduces the expression of pro-inflammatory factors TNF-α,IL-1β and IL-6 in the lumbar spinal cord,and increases the expression of anti-inflammatory factor IL-10,which may be the possibility of electroacupuncture in FM model rats to exert anti-inflammatory effect mechanism. |
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