Screening for blood leukocyte microRNA biomarkers responsible for association between qi deficiency constitution and Pi-qi-deficiency syndrome of chronic superficial gastritis

Abstract

Objective: To screen for blood leukocyte microRNA (miRNA) biomarkers responsible for the association between the traditional Chinese medicine (TCM) qi deficiency constitution (QDC) and the Pi-qi-deficiency syndrome (PQDS) of chronic superficial gastritis (CSG). Methods: Peripheral blood leukocytes were separated from people of two TCM constitutions (balance and qi deficiency) and from CSG patients with PQDS. Total RNA was isolated from the leukocytes and subjected to subsequent high-throughput miRNA sequencing to identify the miRNAs that are specifically and highly expressed in persons of QDC and CSG patients with PQDS. In addition, the target genes of the associated miRNAs were predicted. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway-based enrichment analyses of these target genes were performed to further evaluate the associated miRNA candidates as potential biomarkers responsible for the association between QDC and PQDS of CSG. Results: Compared with the control group with a balance constitution (P 1.5 or <0.5), 31 and 38 differentially expressed miRNAs were found in persons of QDC and CSG patients with PQDS, respectively. In particular, hsa-miR-145-5p and hsa-miR-146a-3p were highly expressed in both the persons of QDC and CSG patients with PQDS. GO analysis of the target genes of the two common miRNAs showed that they were mainly associated with functions related to synthesis and metabolism, such as cellular nitrogen compound metabolic processes, biosynthetic processes, cellular protein modification processes, and cellular component assembly. KEGG analysis identified the common pathways enriched among the target genes, including the Hippo signaling pathway and the transcriptional misregulation pathway. The common target genes of the two miRNAs seemed to be associated with the spliceosome pathway and the RNA degradation pathway. Conclusion: Hsa-miR-145-5p and hsa-miR-146a-3p may serve as candidate biomarkers responsible for the association between QDC and PQDS of CSG. Keywords: Qi deficiency constitution, Pi-qi-deficiency syndrome, Chronic superficial gastritis, MicroRNA sequencing