| Objective To observe the effect of massage on peripheral sensitization and inflammatory factors in FM model rats,and to explore the effectiveness and mechanism of massage in the treatment of fibromyalgia.MethodsThirty-six male SD rats(body weight 150 ± 20 g)were selected.Twelve rats were randomly selected as blank control group(blank group).Another 24 rats were modulated under isoflurane(1%)anesthesia by injecting 2-morphiol ethanolsulfonic acid along the left tibialis anterior muscle on days 0 and 5.On day 0 and 14,behavioral changes of the threshold of mechanical foot retraction reflex were observed to confirm the success of modeling.On day 15,the successful modeling rats were randomly divided into model control group(hereinafter referred to as model group)and massage treatment group(hereinafter referred to as massage group),with 12 rats in each group.Massage group: the rats were fixed on the binding instrument,and wearing a black cloth head cover to ensure that the rats remained in a relatively fixed position and calm state."Zusanli" and "Yanglingquan" were selected on the left side of the rats,and self-made animal experimental massage intensity recording device was used(Patent Number: 202123156152.5),the acupoint massage intervention,intervention intensity of 0.4 kgf(≈4 N),each point 10 min,a total of 20 min,once a day,intervention once every other day,continuous intervention for 7 times,a total of 14 days.Control group and model group: no intervention,only grasping and binding fixation.Mechanical pain threshold was measured in all rats after intervention.Morphological changes of muscle were observed by HE staining.The positive expression of SP,MC and 5-HT in skin was observed by immunofluorescence histochemistry.The protein contents of 5-HT,IL-1β,IL-6,TNF-α and IL-10 in serum and muscle were determined by suspension microarray method.Results1.Determination results of mechanical retraction reflex thresholdBefore modeling(day 0),there was no significant difference in the threshold of mechanical foot contraction reflex among all groups.After the first intramuscular injection of 2-morphiol ethanolesulfonic acid(day 14),the threshold of mechanical foot contraction reflex was significantly decreased in all groups compared with the control group(P<0.05);At the end of intervention(day 28),the threshold of mechanical foot contraction reflex in the massage group was significantly higher than that in the model group(P<0.05).2.Skeletal muscle HE staining results(1)Coronal section of tibialis anterior muscle:Control group: muscle fibers were regular polygons,with abundant sarcomere and sarcoplasm without rupture,and myofibrils were closely arranged.There were multiple tightly attached nuclei on the surface of muscle fibers,and the nuclear membrane was smooth,and the size and color of nuclei were uniform.Model group: muscle fibers had rounded edges and chaotic structure,irregular widening of muscle space and widening of interstitial gap,obvious indention of the nuclei attached to the surface,and the size and color of the nuclei were not uniform.The swollen muscle fibers were enlarged in the middle part of the contracted nodule,and the reduced muscle fibers were reduced in the two ends of the contracted nodule.Massage group: the shape of muscle fibers was regular,the structure was clear,the muscle gap was not significantly widened,a few nuclei were observed on the surface,and the size and color of the nuclei were uniform.(2)Horizontal section of tibialis anterior muscle:Control group: muscle fibers were neatly arranged,coherent up and down,fiber thickness and size were equal,no inflammatory infiltration or abnormal morphological changes were observed.Model group: muscle fiber arrangement was disorder,there were many muscle fiber and muscle segment fracture,muscle fiber thickness and size varied,muscle fiber swelling or degeneration necrosis,obvious inflammatory infiltration and abnormal morphological changes.In the massage group,the muscle fibers were arranged in order,and a small number of muscle fibers and muscle segments were broken.The fiber thickness and size were equal,and the inflammatory infiltration and abnormal morphological changes were less than those in the model group.3.Skin immunofluorescence histochemistry test results(1)Compared with the control group,the positive expression of SP in epidermis and epiderm-dermal junction in model group was significantly increased,with statistical significance(P<0.05);Compared with model group,the positive expression of SP in epidermis and epiderm-dermal junction was significantly decreased in massage group(P<0.05).(2)Compared with the control group,the positive expressions of MC and 5-HT in epidermis and epiderm-dermal junction in model group were significantly increased,and their morphology was increased,with statistical significance(P<0.05).Compared with the model group,the positive expressions of MC and 5-HT in the epidermis and epiderm-dermal junction in the massage group were significantly reduced,and the morphology was reduced,with statistical significance(P<0.05).4.Serum test results by suspension chip method(1)Compared with the control group,the expression level of the neurotransmitter 5-HT in serum of model group was significantly increased,with statistical significance(P<0.05);Compared with model group,the expression level of neurotransmitter 5-HT in serum of rats in massage group was significantly decreased,with statistical significance(P<0.05).(2)Compared with control group,the expression levels of pro-inflammatory factors IL-1β,IL-6and TNF-α in serum of model group were significantly increased,with statistical significance(P<0.05);Compared with model group,the expression levels of pro-inflammatory factors IL-1β,IL-6 and TNF-α in serum of rats in massage group were significantly decreased,with statistical significance(P<0.05).(3)Compared with the control group,the expression level of anti-inflammatory factor IL-10 in serum of model group was significantly decreased,with statistical significance(P<0.05);Compared with model group,the serum expression level of anti-inflammatory factor IL-10 in massage group was significantly increased,and the difference was statistically significant(P<0.05).5.Muscle test results by suspension chip method(1)Compared with control group,the expression level of neurotransmitter 5-HT in muscle of model group was significantly increased,with statistical significance(P<0.05);Compared with model group,the expression level of neurotransmitter 5-HT in muscle of rats in massage group was significantly decreased,and the difference was statistically significant(P<0.05).(2)Compared with control group,the expression levels of pro-inflammatory factors IL-1β,IL-6and TNF-α in muscle of model group were significantly increased,the differences were statistically significant(P<0.05);Compared with model group,the expression levels of pro-inflammatory factors IL-1β,IL-6 and TNF-α in muscle of rats in massage group were significantly decreased,with statistical significance(P<0.05).(3)Compared with the control group,the expression level of anti-inflammatory factor IL-10 in muscle of model group was significantly decreased,with statistical significance(P<0.05);Compared with model group,the expression level of anti-inflammatory factor IL-10 in muscle of massage group was significantly increased,and the difference was statistically significant(P<0.05).Conclusions1.Massage can improve the mechanical foot contraction reflex threshold of FM model rats and relieve pain.2.Massage can reduce the inflammatory mediators in injured skeletal muscle of FM model rats and promote the repair process of injured skeletal muscle.3.Massage can down-regulate the expressions of SP,MC and 5-HT in the skin tissue of FM model rats,reduce the stimulation and release of pain-causing substances in the peripheral system,and relieve peripheral sensitization.4.Massage can down-regulate the expression levels of pro-inflammatory factors IL-1β,IL-6 and TNF-α in serum and muscle of FM model rats,and up-regulate the expression level of anti-inflammatory factor IL-10,change the expression of inflammatory factors in peripheral system,and relieve peripheral sensitization.5.Massage can alleviate the pain caused by peripheral sensitization in FM model rats.This mechanism may be achieved by inhibiting the expression of SP in the peripheral system,reducing the aggregation and release of MC,and then down-regulating the expression levels of5-HT and pro-inflammatory factors,and up-regulating the expression level of anti-inflammatory factors through the mechanical effect of massage. |
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