Effect Of Acupuncture On Differential Protein Expression In Spinal Proteomics Of Rats With Myofascial Trigger Points

Objective:Differential proteins in spinal cord of rats at myofascial trigger points were screened by TMT proteomic analysis.These proteins were annotated by GO function and enriched by KEGG pathway.According to the results of functional annotation of GO and enrichment of KEGG pathway,the signal pathways which related to the pathogenesis of MTr Ps were screened out,and the most related to the pathogenesis of MTr Ps were selected for study.Finding out the function of differential proteins in the selected signal pathway by Unpoit,and exploring how they interact with each other to lead to the formation of MTr Ps in rats' gastrocnemius muscle.Then through Western blotting to observe the changes of Differential proteins' expression after acupuncture and false acupuncture intervention,so as to provide a theoretical basis for the clinical effectiveness of acupuncture in the treatment of trigger pain.Objects and methods :As the model of SD rats,four groups were designed:Control Group(CG),Model Group(MG),Dry Needling of MTr Ps(DG)and Dry Needling of Non-Dry Needling(NDG).The whole experiment was carried out in two steps.Ten rats spinal cord samples from CG group and MG group were selected for TMT relative quantitative proteomic bioinformatics analysis.The differential proteins between the two groups were screened out according to the criteria of multiple change greater than 1.2 times(up-regulated more than 1.2 times or down-regulated less than0.83)and P-value less than 0.05,and then combined according to the results of GO functional annotation and KEGG analysis.The proteins related to the pathogenesis of Myofascial Trigger Points were screened out.Western Blot test was performed to verify the changes of up-regulated or down-regulated protein levels in the spinal cord of the four groups.the data were analyzed by SPSS 20.0 software and single factor analysis of variance(ANOVA).Results1.iTRAQ experimental results :A total of 5095 spinal cord proteins were identified in SD rats.Finally,70 up-regulated proteins and 58 down-regulated proteins were screened.Finally,according to the purpose of the study,we chose the most significant pathway of differential protein enrichment to further study the interaction between ECM-receptors through GO functional annotation and KEGG pathway enrichment.The main differentially expressed proteins in this pathway are THBS-1,COL6 A,ITG?3 and VN.2.The results of Western blot :The expression levels of THBS-1,VN,ITG ? 3and COL6 A in MG group / CG group are increased,and the difference between the two groups was very significant(P < 0.01).There were consistent with the results of iTRAQ experiment.We also found that the expression of COL6 A protein in DG was significantly lower than MG(P < 0.01),and the expression level was also decreased after sham acupuncture intervention(P < 0.05).NDG was not as fast as DG in the decrease of COL6 A expression.The expression of ITG?3 in DG group / MG group and NDG group / MG group decreased significantly(P < 0.01).The DG / MG group of vitronectin decreased significantly(P < 0.05),but there was no difference between DG group and MG group.The expression level of protein THBS-1 was no difference between DG / MG group and NDG / MG group.But,compared with MG group,the expression level of this protein between DG group and NDG group all decreased.Conclusion1.The up-regulation of the expression levels of THBS-1,ITG?3,COL6 A and VN in the spinal cord of rats may promote the occurrence and formation of synapses in central neurons,especially the increase of excitatory synapses,that may be leading to central sensitization in MTrPs rats.2.Acupuncture may reduce the central injury by reducing the up-regulated expression of ITG?3 in the spinal cord of MTr Ps rats.In addition,acupuncture is more effective in accurately locating the position of MTr Ps on skeletal muscle than acupuncture in skeletal muscle beside the trigger point.