Integrated Analyses of miRNA and mRNA Profiles in Leukocytes and Serums in Traditional Chinese Medicine (TCM)-Defined Pi-qi-deficiency Syndrome and Pi-wei damp-heat Syndrome Resulting from Chronic Atrophic Gastritis
Authors: You Leiming , Zhang Shen , Li Ting’an , Sang Xiaopu , Li Kunyu , Wang Wei , Gao Xinhui , Wu Jiarui , Huang Guangrui , Wang Ting , Xu Anlong 
Published: September 2020

Abstract
Background: To investigate the microRNA (miRNA)-gene interactions underlying leukocyte functions and characteristics, especially the potential serum biomarkers, implicated in the traditional Chinese medicine (TCM)-defined Pi-qi-deficiency syndrome (PQDS) and Pi-wei damp-heat syndrome (PDHS) resulting from chronic atrophic gastritis (CAG). Methods: Using RNA/miRNA-sequencing approach, we identified the syndrome-specific miRNAs and genes in leukocytes or serums, and further analyzed their functions and pathways to decode their potential roles in contributing to the characteristics and functions of leukocytes. Especially, we validated the syndrome (Zheng)-specific miRNA-gene interactions to explorer their roles for the characteristic and functional changes of leukocytes. Also, we evaluated the possibility of location of syndrome-specific miRNAs into plasma exosomes, and analyzed their validated targets to reveal their potential roles in the body.Results: Compared with healthy control population, we found that despite being the TCM-defined syndromes resulting from the same disease of CAG, the Zheng-specific genes and miRNAs were not same. The PDHS-specific leukocyte genes were mainly involved in defense and immune responses, and enriched in neutrophil degranulation and nucleotide-binding-oligomerisation-domains (NOD)-like receptor signaling pathways, as well as several synapses-related pathways. The expression upregulation of PDHS-specific genes enriched in the neutrophil degranulation pathway, indicated the enhanced leukocyte degranulation activation in the PDHS. The PQDS-specific genes in leukocytes were implicated in inflammatory response, extracellular matrix (ECM) organization and collagen catabolism. They could be enriched in mitogen-activated protein kinase (MAPK) signaling, IL17 signaling and helper T (Th) cell differentiation pathways, especially the pathways associated with cell-to-cell adhesion/junction and communication such as ECM-receptor interaction, cell adhesion molecules (CAM) and ECM organization, probably directly contributing to the characteristics and functions of leukocytes in the PQDS. Also, the experimentally-supported miRNA-gene interactions, concerned with the targets of COL4A2 (collagen, type IV, alpha 2), COL26A1 (collagen, type XXVI, alpha 1), SPP1 (secreted phosphoprotein 1) and PROCR (endothelial protein C receptor), were specifically implicated in the regulation of pathways related to cell-to-cell adhesion/junction and communication, suggesting the potential roles of the PQDS-specific miRNA-gene interactions for the characteristic and functional changes of leukocytes in the PQDS. Interestingly, the PQDS-specific miRNAs in the serums and the corresponding leukocytes, seemed to have the common roles in contributing to the characteristics and functions of leukocytes in the TCM-defined PQDS of CAG. Importantly, the hsa-miR-122-5p could be a potential biomarker candidate for the TCM-defined PQDS, capable of being contained and carried in plasma exosomes and especially much higher expression in both the leukocytes and corresponding serums in the CAG patients with PQDS rather than PDHS. Conclusions: These results may provide new insights into the characteristic and functional changes of leukocytes in the two TCM syndromes, PDHS and PQDS, especially the miRNA-mediated gene regulation underlying leukocyte characteristics and functions, with potential leukocyte and serum biomarkers for future application in integrative medicine.Trial registration: ClinicalTrials.gov, NCT02915393. Registered on September 17, 2016.