| ObjectiveBy regulating the expression of serum exosome mi R-124,the mechanism of action of fire acupuncture therapy on nerve function recovery after SCI was explored,providing partial experimental basis for promoting fire acupuncture therapy on SCI.MethodsThirty-six healthy female SD rats were divided into 4 groups-blank group randomly,sham group,model group,fire needle group,with 9 rats in each group.Detection was performed at 14 d.SCI model was prepared by Allen’s method.Only the ninth thoracic and the ten thoracic spinous process and partial lamina resection were implemented in the blank group,and grasping and fixation with the same intensity as in the fire needle group were performed at the same time,but no intervention was performed.The model group was given the same intensity of grasping and fixation at the same time as the fire needle group without acupuncture.In the fire needle group,the fire needle was used to puncture the seven thoracic,the eight thoracic,the eleven thoracic,the twelve thoracic acupoints under the spinous process and on both sides of the jiaji acupoints.The fast acupuncture method was used to quickly pierce into the selected acupoints and quickly Pierce out without retention of needles,24h/time.BBB score was used to observe the improvement of nerve dysfunction in SCI model rats by fire acupuncture from the perspective of animal behavior.Transmission electron microscopy,Western-blot and RT-q PCR were used to detect the effects of fire needle therapy on serum exosome morphology and expression of mi R-124.The expression of TLR4,MYD88,NF-κB and Arg-1 in M2 microglia was detected by Western-blot.Results1.Effects on neurological function: BBB scores showed that after modeling,both the model group and the fire needle group showed loss of hind limb motor function,the differences between the two groups were not statistically significant(P=0.551>0.05),indicating that the level of modeling was consistent between the two groups,while no motor function defect was found in the blank group and the sham group(BBB score was 21).After14 days,the model group showed a tendency of motor function recovery,and after 14 days of fire acupuncture intervention,the BBB score of the fire acupuncture group was higher than that of the model group,the difference was statistically significant(P<0.001).2.Effect on the expression of serum exosome mi R-124: According to RT-q PCR results,the relative expression level of mir-124 in exosome of sham operation group and model group was lower than that of blank group on day 14(sham operation group: P<0.001;Model group:P<0.001),there was no significant difference between the sham group and the model group(P=0.561>0.05).Compared with the model group,the relative expression level of mi R-124 in exosomes in the fire needle group was significantly higher than that in the model group,with statistically significant difference(P=0.001).3.Effect on TLR4/NF-κB signaling pathwayWester Blot results showed that 14 days after fire acupuncture treatment:(1)In model group,TLR4 protein expressionwas was higher than other three groups,between blank group and sham group,there was no significant difference(P<0.05),while there was no significant difference between the sham group and the model group(P>0.05).Compared with the model group,TLR4 protein expression in the fire needle group was lower than that in the model group,and the difference was statistically significant(P<0.05);(2)The protein expression level of MYD88 in the fire needle group and the model group was significantly higher than the other two groups,and there was no significant difference between the blank group and the sham group(P<0.05),the difference between the model group and the sham group was statistically significant(P<0.05);Compared with the model group,the expression of MYD88 protein in the fire needle group was lower than that in the model group,and the difference was statistically significant(P<0.05);(3)In the model group,NF-κB protein expression was higher than the other three groups,between blank group and sham group,there was no significant difference(P<0.05);Between model group and sham group,the difference was statistically significant(P<0.05),the relative expression of NF-κB in fire needle group was lower than the model group,and the difference was statistically significant(P<0.05);(4)The expression of Arg-1 protein in the fire needle and the model group was significantly higher than that in the sham group and the blank group,between the sham group and the blank group,there was no significant difference(P<0.05);Between model group and sham group,the difference was statistically significant(P<0.05);Compared with the model group,the relative expression level of Arg-1 in the fire needle group was higher than that in the model group,and the difference was statistically significant(P< 0.05).Conclusion1.Fire needle therapy can effectively protect damaged local neurons in SCI rats,improve the motor function of hind limbs and promote the recovery of nerve function.2.Fire needle therapy can regulate and promote the secretion of serum exosome mi R-124,then inhibit TLR4/NF-κB signaling pathway,promote the polarization of M2 microglia,and play a role in nerve repair. |
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