Objective: To investigate the differential characteristics of plasma mircoRNA (miRNA) expression profile in the patients of moderate-to-severe allergic rhinitis treated with acupuncture so as to provide an index for screening the potential biomarkers of acupuncture efficacy.
Methods: Of 33 patients of moderate-to-severe allergic rhinitis underwent acupuncture, the superior efficacy patients (superior efficacy group, 3 cases) and the inferior efficacy patients (inferior efficacy group, 3 cases) were selected. Using human miRNA microarray technology, the differences in plasma miRNA expression before and after treatment were analyzed in the patients of two groups. Besides, 10 cases of superior efficacy and 10 cases of inferior one were selected respectively among the patients of moderate-to-severe allergic rhinitis treated with same acupuncture regimen; and the real-time PCR was used to validate miRNAs of differential expression determined by microarray technology. The bioinformatics analysis was performed for miRNAs of significant differences in expression so as to predict the potential functional target genes, and then, the predicted target genes were annotated in reference with the databases of gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG).
Results: Before treatment, there were 51 miRNAs of differential expression between two groups, of which, the expression levels of 26 miRNAs were up-regulated and those of 25 miRNAs were down-regulated. Compared with before treatment, 33 miRNAs presented differential expression in the superior efficacy group after treatment. The results of real-time PCR showed that the expression levels of hsa-miR-126-3p, hsa-miR-15a-5p, hsa-miR-494-3p and hsa-miR-574-5p were consistent with the results of microarray analysis in tendency. GO/KEGG analysis indicated that miRNAs with significant differences of expression between two groups were involved in regulating various biological processes, molecular functions and signaling pathways.
Conclusion: Plasma miRNA-mediated biological processes may be associated with the efficacy response of acupuncture in treatment of moderate-to-severe allergic rhinitis. Plasma miRNAs of differential expression may be the potential non-invasive biomarkers to predict the effectiveness of acupuncture on moderate-to-severe allergic rhinitis.
目的：分析针刺治疗中重度变应性鼻炎（AR）不同疗效患者血浆微小核糖核酸（miRNA）表达谱差异特征，为筛选针刺疗效可能生物标志物提供依据。方法：从33例接受针刺治疗的中重度AR患者中选择优效患者（优效组）与劣效患者（劣效组）各3例，采用人miRNA芯片检测技术分析两组患者治疗前后血浆miRNA的表达差异。另选择接受相同针刺治疗的中重度AR患者优效、劣效各10例，采用实时荧光定量PCR对芯片检测中差异表达的miRNA进行验证。对表达差异显著的miRNA进行生物信息学分析，预测其可能作用的靶基因，并利用基因本体（GO）及京都基因与基因组百科全书（KEGG）数据库对预测的靶基因进行注释。结果：共筛选出治疗前两组患者间存在差异表达的miRNA 51个，其中26个miRNA表达上调，25个miRNA表达下调；与治疗前比较，优效组治疗后有33个miRNA存在差异表达；实时荧光定量PCR验证结果示hsa-miR-126-3p、hsa-miR-15a-5p、hsa-miR-494-3p、hsa-miR-574-5p的表达与芯片检测结果趋势一致。GO/KEGG分析显示，两组间表达差异显著的miRNA参与调节多种生物学过程、分子功能和信号通路。结论：miRNA介导的多种生物学过程可能与针刺治疗中重度AR疗效应答有关，差异表达的miRNA有潜力作为预测针刺治疗AR有效性的非侵入性生物标志物。.
Keywords: acupuncture; allergic rhinitis, moderate-to-severe; bioinformatics; efficacy predication; mircoRNA (miRNA) expression profile.