| ObjectiveCompared with healthy individuals,the mRNA and miRNA expression levels of peripheral blood leukocytes and the circulating miRNA expression levels of peripheral blood serum in patients with chronic atrophic gastritis(CAG)were analyzed by high-throughput sequencing,in an attempt to explore the probable biological basis of Pi-Qi deficiency syndrome(PQDS)and Pi-Wei dampness and heat syndrome(PDHS)as defined by traditional Chinese medicine.MethodsAfter a series of diagnostic criteria,inclusion criteria and exclusion criteria screened in Dongfang Hospital of Beijing University of Traditional Chinese Medicine,we recruited 5 healthy subjects,5 CAG patients with PQDS and 5 CAG patients with PDHS.The peripheral blood of all subjects in the above three groups were collected for the separation of leukocytes and sera.Then the total RNA of leukocytes and sera were extracted for high-throughput sequencing.The differential expression of miRNAs and mRNAs/genes of leukocytes and sera in the two syndromes groups were screened by comparing with healthy individuals control.The database was used to predict target genes of the known differentially expressed miRNAs.And KEGG pathway and GO biological function analysis of the predicted target genes were analyzed for the two syndrome-specific differential miRNA of leukocytes and sera.We explored the functions of target genes of the common differential miRNA in the leukocytes and sera under the same syndrome.Then we constructed a relational network of syndrome-specific differential miRNA-differential gene.ResultsAmong the leukocytes of the patients,compared with the healthy control,CAG patients with PDHS were screened out with 24 syndrome-specific differential miRNAs and 145 syndrome-specific differential genes,while CAG patients with PQDS were screened out with 66 syndrome-specific differential miRNAs and 155 syndrome-specific differential genes.KEGG pathway enrichment analysis showed that the PDHS-specific leukocytes differential miRNAs were involved in the extracellular matrix receptor interaction pathway,while the PQDS-specific leukocytes differential miRNAs were not only involved in the extracellular matrix receptor interaction pathway,but also related to fatty acid biosynthesis,Hippo signaling pathway and other pathways.However,the PDHS-specific leukocytes differential genes were related to serotonergic and glutamatergic synapses,and the PQDS-specific leukocytes differential genes were concerned with extracellular matrix receptor interaction pathway,MAPK signaling pathway and fatty acid metabolism.The results of GO function analysis showed that the PDHS-specific leukocytes differential miRNAs were related to the negative regulation of apoptotic process,while the PQDS-specific leukocytes differential miRNAs were related to the biological processes such as neurotrophin TRK receptor signaling,macromolecular complex assembly,cellular protein modification and small molecule metabolism.Furthermore,both group for syndrome-specific leukocyte differential miRNAs were involved in the processes such as gene expression,cell death,catabolism,and cellular protein modification.The PDHS-specific leukocyte differential genes were involved in immune and innate immune response process,while the PQDS-specific leukocyte differential genes were mainly related to the processes of inflammatory response,collagen catabolic and extracellular matrix organization.Among the sera of the patients,compared with healthy control,CAG patients with PDHS were screened out with 37 syndrome-specific differential miRNAs,while CAG patients with PQDS were screened out with 84 syndrome-specific miRNAs.KEGG results revealed that the PDHS-specific and PQDS-specific serum differential miRNAs were related to the pathways of fatty acid biosynthesis,lysine degradation and proteoglycans in cancer.Moreover,the PDHS-specific serum differential miRNAs were much more involved in other cancer-related pathways,and the PQDS-specific serum differential miRNAs were closely related to pathways of fatty acid metabolism,foxO signaling,p53 signaling and transcriptional mis-regulation in cancer.The results of GO function analysis,however,revealed that the biological process enriched by target genes of serum-specific differential miRNAs was consistent with that of leukocytes-specific differential miRNAs.The results of the syndrome-specific differential miRNAs-differential genes relational network showed that the differential target genes regulated by PDHS-specific leukocytes differential miRNAs were related to metabolic process,and the differential target genes regulated by PQDS-specific leukocytes differential miRNAs were mainly related to the immune,cancer and metabolic processes.It was find that differentially expressed hsa-miR-122-5p appeared both in leukocytes and sera in CAG patients with PQDS.But there was no common expressed differential miRNA in leukocytes and sera in CAG patients with PDHS.Predicted target genes of hsa-miR-122-5p were differentially expressed in leukocytes,including JAG2,SH3RF1,MYOF,PCSK6 and COL4A2.Besides,KEGG pathway analysis enriched by the target genes of hsa-miR-122-5p was mainly related to various cancer pathways and PI3K-Akt signaling pathwayConclusionsThe differential miRNAs of leukocytes and sera in CAG patients with PDHS and PQDS were related to metabolism,which may be the potential biological basis of Pi(spleen)deficiency and dysfunction in traditional Chinese medicine.Differential miRNAs of leukocytes and sera in the two syndrome groups were also associated with cancer,which also confirmed that CAG is a manifestation of precancerous lesions.Furthermore,the differential miRNAs and differential genes of leukocytes in CAG patients with PQDS were more related to immunity,cellular adhesion/junction and communication,which may reflect the changes of leukocytic function and characteristics in CAG patients.Especially,the expression of hsa-miR-122-5p of leukocytes and serum in CAG patients with PQDS was high level,comparing with healthy control.The hsa-miR-122-5p was concerned with a variety of cancer pathways and immune-related pathways.It was speculated that hsa-miR-122-5p may be a potential biomarker of PQDS in CAG.But this conjecture needs further exploration. |
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