Effect Of MicroRNA On Deficiency Of Kidney Yang And Intervention Effect Of Yougui Pill

Objectives:The differentially expressed miRNAs of kidney yang deficiency syndrome and Yougui pills before and after treatment were screened by miRNA expression profiling.by predicting and functional analysis of differentially expressed miRNAs target genes,the molecular mechanism of kidney yang deficiency syndrome and the repairing effect of yougui pills were preliminarily explored from the perspective of post-transcriptional regulation of miRNA genes.Method:1.Based on the Chinese knowledge network and kidney-yang deficiency syndrome,and combined with traditional Chinese medicine classic books to study the understanding of this TCM syndrome.the theoretical study of miRNA was carried out by searching the database of foreign literature such as pubmed and geenmedical.2.Using gene chip technique to screen differentially expressed mi RNAs:According to the syndrome differentiation criteria of kidney yang deficiency syndrome,the test subjects were selected,and the total RNA,of peripheral venous blood was extracted for sample labeling and gene chip hybridization.Using multiple analysis method,Fold change≧2 or Fold change≦0.5 and P-value<0.05 as screening conditions,Different expression mi RNAs of healthy control group,kidney-yang deficiency group and Yougui pill group were selected respectively.The miRNAs,relative differential expression between groups were selected by comparing differential expression miRNAs between each of the three groups.3.The screening principle of kidney yang deficiency syndrome related miRNA:the kidney yang deficiency syndrome group and the normal control group compared to obtain the kidney yang deficiency syndrome related differential expression miRNAs,after the treatment of kidney yang deficiency syndrome related differential expression of mi RNAs screening whether there are significant changes in this batch of differential expression miRNAs after treatment can be considered to be closely relatedto kidney yang deficiency syndrome.further screening of miRNAs closely related to kidney yang deficiency using waynchart.4.Target gene prediction of miRNAs closely related to kidney yang deficiency was performed using TargetScanHuman7.2,miRDB,miRTarbase,and target gene GO enrichment and KEGG pathway analysis using DAVID software.5.The results of miRNA expression profiling chip analysis were verified:real-time quantitative PCR detection was carried out by screening the significant miRNAs,and whether the results were consistent with the gene chip results were analyzed.Result1.The differential expression of miRNAs was screened by gene chip technique,of which there were 46 different miRNAs in the kidney yang deficiency syndrome group and the healthy control group,of which 28 were up-regulated and 18 were down-regulated,433 were different miRNAs in the treatment group of kidney yang deficiency syndrome group and Yougui pills group,of which 149 were up-regulated and 284 were down-regulated,and 429 were different miRNAs in the treatment group and the healthy control group,of which 160 were up-regulated and 269 were down-regulated.2.On the basis of the screening principle of the related miRNA of kidney yang deficiency syndrome,19 different expression miRNAs.were,which were closely related to deficiency of kidney yang.The 19 miRNAs were intersected in the TargetScanHuman7.2、mi RDB、miRTarbase and 14 miRNAs.with common target genes were selected.3.using DIVID software to carry out GO enrichment and KEGG pathway analysis of all their common target genes,it was found that the target gene function mainly includes three categories: immune regulation,cell cycle regulation and metabolic regulation.On the basis of the negative regulation effect of miRNA on its target gene,the miR-214-3p function is related to immunomodulation,and the miR-5009-3p、miR-3658、miR-214-3p and miR-4795-5p function is related to cell cycle regulation.mi R-214-3p 、 miR-3658 function is associated with metabolic regulation.4.The real-time quantitative PCR verification results of chip results found that miR-214-3p was differentially expressed in kidney yang deficiency syndrome.conclusion1.miRNA participate in the regulation of the kidney yang deficiency syndrome,the regulatory effect of mi RNA on kidney yang deficiency syndrome is extensive,mainly involving three aspects: immune regulation,cell cycle regulation and metabolic regulation,and Yougui Pill can regulate the syndrome by changing the expression of these characteristic miRNA.2.The miR-214-3p expression of kidney yang deficiency syndrome and Yougui expression miRNA of the curative effect of Yougui pills is significant,and its role in the deficiency syndrome of kidney yang needs further study.