| Objective:1.explore the microscopic differences of miRNA gene expression profiles in salivary glands of Yin deficiency constitution and Yang deficiency constitution,screen molecular markers of these two kinds of biased constitutions,and explore the new direction of objective study of constitution.2.analyze the differences of miRNA gene expression profiles among these two kinds of biased constitutions and Pinghe constitution.Provide a new entry point for improving constitutions to reverse the unhealth constitutions,diagnosis and prevention of diseases.Methods:1.In this experiment,5 volunteers were recruited to the Pinghe constitution group,6 volunteers in the absence of illness to the yin deficiency constitution group,and 5 volunteers in the absence of illness to the yang deficiency constitution group.We got 16 volunteers of different ages and occupations.Gargle with sterile water to remove foreign bodies in the mouth,and collect saliva from the mouth.Isolate the RNA from oral saliva,and detect the human miRNA onearray high-throughput gene chip.2.Analyze miRNA expression profile data,and screen the miRNA with differential expression.3.Do the analysis of target genes in differentially expressed miRNA.And do the analysis of the miRNA target genes to predict the bio-functions of the different miRNAs.Results:1.We found that there were 81 differentially expressed miRNAs between Yang deficiency constitution group and Pinghe constitution group,including 33 up-regulated differential miRNAs and 48 down-regulated differential miRNAs,We found that there were 98 differentially expressed miRNAs between Yin deficiency constitution group and Pinghe constitution group,including 35 up-regulated differential miRNAs and 63 down-regulated differential miRNAs,We found that there were 167 differentially expressed miRNAs between Yang deficiency constitution group and yin deficiency constitution group,including 51 up-regulated differential miRNAs and 116 down-regulated differential miRNAs,2.through KEGG and GO enrichment analysis of differentially expressed miRNA target genes,we found that there were abnormal expressions of biological pathways such as hormone metabolism,Glycolipid,immune regulation,tumor between Yang deficiency constitution group and Pinghe constitution group,The abnormal expressions of inflammatory reaction,immune regulation,tumor and other biological pathways were found between yin deficiency constitution group and Pinghe constitution group,Abnormal expressions of immune regulation,hormone metabolism,tumor and other biological pathways were found between yang deficiency constitution group and yin deficiency constitution group.Conclusion:1.There was a systematic difference in the expression of miRNA at the whole genome level in the patients between Yang deficiency constitution group and Pinghe constitution group,Yin deficiency constitution group and Pinghe constitution group,Yang deficiency constitution group and yin deficiency constitution group.2.through the analysis of target gene KEGG we found that there were differences in the immune pathway between Yin deficiency constitution group and Pinghe constitution group,so as differences between Yang deficiency constitution group and Pinghe constitution group.It showed that the two kinds of biased constitution may have the molecular mechanism of abnormal immunological pathway at the gene level.At the same time,there were different expressions of thyroid hormone,male hormone and other related hormone pathways in Yang deficiency constitution group compared with Pinghe constitution group.There were different expressions of inflammatory pathway,hormone related pathway in Yin deficiency constitution group compared with Pinghe constitution group.That may be the internal cause of weak Yang features in Yang deficiency constitution and weak Yin features in yin deficiency constitution.3.We chose human oral saliva as the research object,the mirna expression profiling as the research method.And the expression of miRNA in saliva of yin deficiency constitution and yang deficiency constitution was preliminarily constructed.We screened differentially expressed target genes,analyzed KEGG and GO enrichment results,laid the foundation of the molecular mechanism of abnormal performances on yin deficiency constitution and yang deficiency constitution. |
