Study On The Regulation Mechanism Of MicroRNA Expression In IBS-D Patients Treated With Herb-partitioned Moxibustion On The Navel

Objective:To explore the regulation mechanism of miRNA expression in IBS-D patients treated with Herb-Partitioned Moxibustion on the navel,and to clarify the partial target group of this therapy at the level of miRNA,so as to provide evidence for the regulation of gene expression in the treatment of IBS-D patients with Herb-Partitioned Moxibustion on the navel.Methods:1.The 20 patients diagnosed IBS-D were assigned to the Herb-Partitioned Moxibustion on the navel group,and 10 healthy volunteers matched in age and gender were assigned to the normal control group to detect the differential expression profiling of miRNA in peripheral blood of the Herb-Partitioned Moxibustion on the navel group before treatment and the normal control group.2.The patients diagnosed IBS-D were treated with Herb-Partitioned Moxibustion on the navel,once a week for 2 hours each time,4 times as a course of treatment,2consecutive courses of treatment,a total of 8 times.The IBS-SSS scale was used to evaluate the therapeutic effect,and the differential expression profiling of miRNA in peripheral blood of IBS-D patients were detected before and after treatment.3.To predict target gene using miRWalk3.0 database and miRDB6.0 database.Then taking the intersection of two databases and carrying out both GO enrichment analysis and KEGG pathway analysis in order to explore the biological function of target genes.4.RT-PCR was used to detect the levels of differentially expressed miRNA in the serum of 20 IBS-D patients and 10 healthy controls,so as to verify the microarray results and explore the influence of Herb-Partitioned Moxibustion on the navel on the blood miRNA expression of IBS-D patients.Results:1.Microarray results showed that there were significant differences in the expression of 5 miRNAs in peripheral blood of IBS-D patients before treatment compared with the normal control group,which hsa-let-7b-5p,hsa-mir-19b-3p,hsa-mir-20a-5p,hsa-mir-1246 were down-regulated and miRNABright Corner30 was up-regulated.After treatment,there were significant differences in the expression of 5 miRNAs in peripheral blood of IBS-D patients compared with before treatment,namely,hsa-let-7b-5p,hsa-mir-19b-3p,hsa-mir-20a-5p,hsa-mir-16-5p and hsa-mir-630,which were all up-regulated.2.Clinical symptoms of IBS-D patients were intensely improved by the intervention of Herb-Partitioned Moxibustion on the navel,which IBS-SSS score was reduced from236.85±49.24 points before treatment to 95.90±37.05 points with statistical significance(P<0.01)and the effective rate was 90.00%.3.Bioinformatics analysis results showed that 387 target genes were found in the group before treatment/normal control,and 487 in the group after/before treatment.GO enrichment and KEGG pathway analysis were performed on the predicted target genes.It was found that the function of target genes was related to cell adhesion,nervous system development,posttranscriptional gene silencing by RNA,miRNA mediated inhibition of translation,cadmium ion binding,GTPase binding,voltage-gated sodium channel activity,prolactin signaling pathway,neurotrophin signaling pathway,etc.4.RT-PCR results showed that the expressions of hsa-let-7b-5p,hsa-mir-19b-3p and hsa-mir-20a-5p could be up-regulated after the treatment of the Herb-partitioned moxibustion on the navel(P<0.05).Conclusions:1.It was confirmed that the expression of miRNAs in peripheral blood of IBS-D patients before treatment was significantly different from that of the control group,which may provide data support for the diagnosis and treatment of IBS-D.2.Herb-partitioned moxibustion on the navel has a significant effect on IBS-D.3.Herb-partitioned moxibustion on the navel may achieve therapeutic effects by up-regulating the expression of hsa-let-7b-5p,hsa-mir-19b-3p and hsa-mir-20a-5p,providing evidence for the regulation of gene expression in the treatment of IBS-D.