Study On The Correlation Between The Clinical Effect Of Acupuncture In The Treatment Of Chronic Fatigue Syndrome And The Regulation Of MicroRNA

Objects:1.To fliter out discrepant circulating miRNA in CFS and healthy participants and search the mi RNA linked with onset and progression of CFS by the means of high-throughput sequencing2.To explore the correlation between clinical benefits of acupuncture and targeted effects on miRNA of acupuncture in CFS Methods:1.We ploted discrepant expression profile of miRNA,tested by high-throughput sequencing technology,by comparing fifteen CFS patients circulating miRNA to eight healthy controls datas.Basing on the principle that high difference mutiple,stable expression,we oppositely derivated discrepant expression miRNA from target genes in involved pathway enrichment.The functional classification of target genens,extracted by RNAhybrid 、 miRanda and TargetScan techaniques,was analyzed with GO significant enrichment analysis in on Gene Ontology database.Besides,The Pathway significant enrichment analysis that took KEGG pathway as unit was conducted to identify the biological pathways of target genes,the results were verified by RT-qPCR.2.Of the 15 randomly assigned patients,the 8 in acupuncture group(A group)and7 in placebo acupuncture(B group).Acupoints,GV20,HT7,BL18,BL20 and BL23,were acupunctured for the treatment of A group,and 6 non-channel –non-acupoints were acupunctured with Streitberger trochar for B group.Sessions for two groups took place 5 times a week for preceding 2 weeks of 4 weeks and 3 times a week for remaining 2 weeks.The outcome was the change in Chalder Fatigue Scale and SPHERE scale at baseline and the end of 4 weeks.Meaniwhile,the expressions of circulating miRNA in 4 weeks in A group,B group and control group were estimated by real time quantitative PCR(RT-qPCR),and the correlation between clinical benefits of acupuncture and targeted modulation on miRNA of acupuncture in CFS wasconducted by correlation analysis.Results:1.Microarray analysis showed that,compared with the matched healthy control,84 mi RNAs expressions were significantly different(folds ≥2,Q < 0.05),of which 48 miRNAs were up-regulated while 36 miRNAs were down-regulated in the CFS patients’ serum.18 mi RNAs were newly discovered(folds ≥2,Q<0.05),one of them were down-regulated and the left were up-regulated.RT-qPCR confirmed that the expression of hsa-miR-1-3P in CFS was significantly lower(P < 0.05),the expression of novel_mir122 and novel_mir347 increased significantly(P < 0.05),consistent with the results of high-throughput sequencingA total of 16087 target genes were regulated according to predilection,and 9575 of them were consolidated by exception of corresponding screening like free energy and score values.The target genes were enriched in cellular processes,development and metabolic processes of single organism,protein kinase activity and other biological processes.And 30 related biological pathways were mainly enriched in signal transduction pathways(MAPK signaling pathway,Ca2+ signaling pathway,Wnt signaling pathway),and Natural killer cell mediated cytotoxicity,purine metabolism and other signaling pathway(P<0.05).2.(1)At 4 weeks,the A group had a significantly greater decrease in the total score,scores on mental and physical components of Chalder Fatigue Scale than did the B group(P<0.05)(2)At 4 weeks,A group had a significantly greater increase in hsa-miR-1-3P than B group(P<0.05)(3)At 0 week,Chalder Fatigue Scale total score negatively correlated with hsa-miR-1-3P expression and positively with novel_mir122expression(P<0.05),and mental component score of Chalder Fatigue Scale positively correlated with novel_mir347.At 4 weeks,changes in Chalder Fatigue Scale total score and mental component score of Chalder Fatigue Scale positively correlated with decrease in novel_mir122 for A group when compared with B group(P<0.05).Conculsions:1.CFS patients showed significant elevated expressions in novel_mir122 and novel_mir347 and declined in expressions of hsa-miR-1-3P in peripheral blood.The abnormal changes in miRNA might target on immune inflammation,oxidative and nitrative stress,and energy metabolism pathways,and further modify the onset and progression of CFS2.Acupuncture indicated benefits on symptoms as fatigue,nervous or anxious feeling and attentional impairment.This may result from the effects benefits of acupunture on immune inflammation,oxidative and nitrative stress,and energy metabolism pathways that were originally linked with decrease in novel_mir122.