| ObjectiveInvestigate the mechanism of fire needle acupuncture to improve neuromotor function in rats with spinal cord injury(SCI)by observing the role and targets of fire needle therapy in regulating nerve cell apoptosis.Methods108 female SD rats were randomly divide into sham operation group,model group and fire needle group,and each one was spanided into four time points of 1,3,7 and 14 days.In the sham operation group,only the T9 and T10 spinous processes and parts of the vertebral plate were removed.In the model and sham operation group,no intervention was made.In the fire needle group,the T7,T8,T11 and T12 and points of 0.5 cm close to them punctured with fine fire needles daily by the fast-pricking method.The BBB score was used to observe the improvement of neurological dysfunction in SCI rats by fire needle therapy.TUNEL staining was used to observe the effect of fire needle therapy on neuronal apoptosis,EB staining to observe the effect of fire needle therapy on the change of blood-spinal cord barrier(BSCB)integrity in SCI rats,and Western blot to detect the effect of fire needle therapy on the expression of apoptosis-related proteins(TGF-β,DAP12,p-PI3K).Results(1)Behavioral scoring results: the rats in the sham operation group were 21±0.00 at all time points,and the BBB scores of rats in the model group were greatly lower than those in the sham operation group at 1d,3d,7d,and 14 d after spinal cord injury,and the differences were statistically significant(1d: P<0.001;3d: P<0.001;7d: P<0.001;14: P<0.001);after fire needle intervention,the BBB scores gradually increased at 1d,3d,7d,and 14 d,and all of them were greatly higher than the model group(1d: P=0.004<0.05,3d: P<0.001,7d: P<0.001,14d: P<0.001).(2)The EB staining results showed: there was no or only a small amount of EB exudation from the BSCB at all time points in the sham operation rats.On the comparation of the sham operation group,the EB content of rats in the model group increased at all four time points,and the differences were statistically significant at 1d,3d and 14d(1d: P=0.045<0.05;3d: P=0.031<0.05;14d: P=0.009<0.05);compared with the model group,the EB content of rats in the fire needle group decreased at all four time points,and the differences at 14 d were statistical significance(P=0.031<0.05).(3)TUNEL staining showed: apoptotic nerve cells were occasionally seen in the sham operation group at all time points,while a large number of apoptotic cells were seen in the model group at all time points,and the apoptosis of neurocyte in the fire needle group was significantly less than that in the model group.Comparison of apoptotic indices showed that the apoptotic indices of rats in the sham operation group were lower at four time points,and no significant changes were seen at four time points.The apoptosis index was greatly more in the model group than the sham operation group at all time points(1d: P=0.036<0.05;3d:P=0.017<0.05;7d: P<0.001;14d: P<0.001).After fire needle intervention,the apoptotic index was significantly lower than that in the model group at all time points(1d:P=0.036<0.05;3d: P=0.017<0.05;7d: P<0.001;14d: P<0.001).(4)Western blot results showed that: TGF-β protein expression level was higher in the model group at 1d,7d,and 14 d,The contrast of the sham operation group and model group was not statistically significant only at 14d(P=0.036<0.05);TGF-β protein expression in the fire needle group was higher than that in the model group at 3d and 7d,but the difference was statistically significant only at 3d(P<0.001).The relative expression of DAP12 protein in the model group tended to increase at all four time points by contrast with the sham group,but the differences were not statistically significant;the relative expression of protein in the fire needle group tended to increase at all four time points by contrast with the model group,but the differences were not statistically significant.On the comparation of the sham operation group,the p-PI3 K protein relative expression in the model group tended to increase at all four time points,but the difference was statistically significant only at 1 d(P=0.036<0.05);compared with the model group,the p-PI3 K protein relative expression in the fire needle group was greatly more at 7 d,and the difference was statistically significant(P=0.006<0.05)ConclusionsFire needle therapy could promote neurological recovery in SCI rats,and the mechanism might be related to the regulation of TGF-β,DAP12,and p-PI3 K proteins,which in turn reduces neuronal apoptosis and improves BSCB integrity. |
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