脾虚证患者血清微RNA表达谱的筛选与生物信息学分析

目的 筛选脾虚证患者血清miRNA表达谱进行生物信息学分析，从miRNA水平探讨脾虚证的发病机制和辨证分型规律。方法 选择高脂血症脾虚证和脾胃湿热证患者各4例，以及健康志愿者5名，提取其血清RNA，进行miRNA定量PCR芯片检测。筛选脾虚证患者血清miRNA表达谱，进行层次聚类、靶基因预测和功能注释等生物信息学分析。结果 筛选出脾虚证患者9个候选血清miRNA，其中6个表达上调，3个表达下调；候选miRNA的表达在脾虚证患者、脾胃湿热证患者及健康志愿者3组样本间存在差异。京都基因与基因组百科全书（KEGG）通路分析结果显示，6个上调miRNA调控的83个靶基因显著富集于7个通路，主要涉及细胞因子-细胞因子受体相互作用通路、病原虫感染性疾病、免疫/炎症相关的信号通路和胰腺癌等；3个下调miRNA调控的365个靶基因显著富集于5个通路，主要涉及神经营养因子信号通路、磷脂酰肌醇信号通路、RNA运输、磷酸肌醇代谢和氨基酸代谢等。结论 本研究筛选出的9个脾虚证患者候选miRNA可作为脾虚证临床辨证的潜在血清标志物，为脾虚证患者发病机制的认识和研究提供依据。

Screening and bioinformatic analysis of serum microRNA expression profiles in patients with Pi-deficiency syndrome

Objective To explore the pathogenesis and syndrome differentiation of Pi-deficiency syndrome (PDS) from microRNA (miRNA) levels through screening and bioinformatic analysis of serum miRNA expression in PDS patients. Methods Four hyperlipemia patients with PDS, 4 hyperlipemia patients with Pi-Wei damp-heat syndrome (PWDS) and 5 healthy volunteers were recruited. Their serum RNA was used in miRNA quantitative PCR array experiment. Serum miRNA expression profiles in PDS patients were screened to perform bioinformatic analysis. Results Nine candidate miRNAs (6 upregulated and 3 downregulated) were screened from PDS patients. These miRNAs were able to clearly distinguish among PDS patients, PWDS patients and healthy volunteers. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed 83 target genes controlled by 6 up-regulated miRNAs were significantly enriched in 7 pathways, which were mainly involved in cytokine-cytokine receptor interaction, pathogens of infectious diseases, immune/inflammatory-related signaling pathway and pancreatic cancer; and 365 target genes controlled by 3 down-regulated miRNAs were significantly enriched in 5 pathways, which were mainly involved in signaling pathways of neurotrophin and phosphatidylinositol, RNA transport, and metabolisms of inositol phosphate and amino acid. Conclusion Our findings provide potential miRNA biomarkers for clinical syndrome differentiation of PDS patients, as well as information for understanding and studying the pathogenesis of PDS patients.