Regulation Of Chaihuang Weikuining On Differential Expression MiRNA In Gastric Ulcer Rat Model With Liver Depression And Spleen Deficiency Research

Objective:1.To establish gastric ulcer rat model with liver depression and spleen deficiency with methods of muti-factors and acetic acid injection;2.To explore the differential expression of miRNA in gastric ulcer rats with liver depression and spleen deficiency,and explore the regulation of Chaihuangweifuning on miRNA further more;3.To explore the possible pathogenesis of disease and mechanism of CHaihuang Weikuining from the molecular level,by the bioinformatics analysis of differential expression miRNAs.Methods:1.Establish gastric ulcer with liver depression and spleen deficiency rat model with methods of multi-factors combining,based on TCM etiology and with acetic acid injection.2.The male SPF SD rats were randomly divided into normal control group,gastric ulcer model group with liver depression and spleen deficiency group,Chaihuang Weikuining(CW)group and Huangqi Jianzhong Decoction(HJD)group,with 8 rats each group.After the intervention for 21 days continuously,the drug groups began to medicine.Then the enzyme linked immunosorbent assay(ELISA)was applied to collect the expression of serumamylase activity(AMY),stomach gastrin-releasing(GAS),plasma serotonin(5-HT),and norepineohrine(NE),to check and evaluate the model.3.Micro RNA Microarray was used to detecting miRNAs’ expression level of each sample(based on the miRNA in Sanger miR Base database 21.0);Screening miRNAs which are differential expressing by differential expression analysis.Continue target genes predicted and bioinformatics analysis of differential miRNAs by Target Scan、miRanda database.Results:1.Compared with the normal control group,the serum AMY,GAS in model group were significantly decreased(P<0.01),and plasma NE level was increased(P<0.01).Compared with the model group,serum AMY,GAS were increased,while plasma NE was decreased in Huangqi Jianzhong Decoction(HJD)and Chaihuang Weikuning(CW).2.Microarray Chip results reveal 94 miRNAs have changed in Model Rats,among which 40 miRNAs were up-regulated and 54 miRNAs were down-regulated,and the family of let-7 and miR-30 are widely involved in the disease.rno-miR-122-5p 、 rno-miR-3084a-3p 、 rno-miR-195-5p 、 rno-miR-375-3p 、rno-miR-181a-5p、rno-miR-30c-5p、no-miR-429、rno-let-7i-5p、rno-miR-24-3p、rno-miR-199a-3p and other miRNAs are co-regulated by HJD and CW,and miRNAs specially regulated by HJD are related to immune and cancer,while CW are related to oxidative stress and cancer.3.Predicting target genes of miRNAs which significantly regulated in model and HJD/CW,their functions were enriched on nucleotide binding、metal ion binding、ATP binding 、 protein binding;And taking part in regulation of transcription,DNA-templated 、 positive regulation of transcription from RNA polymerase II promoter、transmembrane transport.KEGG pathways were enriched on Metabolic pathways、Pathways in cancer、MAPK signaling pathway、Cytokine-cytokine receptor interaction、Chemokine signaling pathway.Conclusions:1.Model rats with gastric ulcer of liver depression and spleen deficiency syndrome were set up successfully,and multi-factors combined acetic acid applied in the research to establish gastric ulcer with liver depression and spleen deficiency rat model were proved to be stable and reliable.2.Expression of miRNAs in model rats has changed and CW can call-back the differential miRNAs,play the therapeutic role through multiple targets and multiple pathways.3.There are differences in miRNAs regulated by HJD and CW,and CW correspondents to the syndrome of liver depression and spleen deficiency.