| Objective:Qn the basis of previous experimental study,we established a rat model of chronic stress gastric ulcer(CSGU)with stagnation of Qi in liver and stomach.i TRAQ technique was used to investigate the mechanism of action of Ganwei Baihe decoction on gastric mucosa of rats with chronic stress gastric ulcer of liver-stomach’qi stagnation type.Method:Thirty four SD rats were randomly divided into three groups: Blank Group(BG),Model Group(MG)and Traditional Chinese medicine group(TG).The Rats in the BG were fed normally.The Rats in the TG and MG were stimulated with five unpredictable factors to establish the model of CSGU with stagnation of Qi in liver and stomach.Normal saline(0.9%)and decoction of Ganwei Baihe decoction were given to rats in MG and TG respectively by the method of gastric lavage,and the stimulation above was continued during this process.The time of both gavage and modeling was 14 days,and samples were taken on the 36 th day.Body weight and open-field-test’scores were measured on the first,21 and 35 day respectively.Calculating gastric emptying rate,ulcer index and ulcer inhibition,examining the anatomy and pathology of gastric mucosa,determining the5-hydroxytryptamine level in serum by enzyme linked immunosorbent assay(Elisa)and examining the gastric mucosa proteomics of three groups by i TRAQ technique,then western-blot was used to verify the results of that.Results:1.Model preparation:(1)General situation: After model making,rats’ mental state,intake of feed and water,activity and stool condition were poor than that of the BG;(2)Anatomy and pathology: Under Light Microscope,the gastric mucosa of the model rats was obviously hyperemic and edematous,which were scattered in punctiform and linear ulcer.The inflammatory cells were infiltrated.(3)Body weight,open-field-test score and gastric emptying rate in MG are lower than those in BG(p < 0.01);(4)Ulcer index and 5-HT level in MG were higher than those in BG(p < 0.01).2.The therapeutic effect of Ganwei Baihe decoction on model rats of CSGU with stagnation of Qi in liver-stomach: After the treatment,(1)the general condition of TG was improved;(2)Body weight:The mean and median weight of the rats in TG were higher than those in MG,but compared with the MG,there was no significant difference(p>0.05)(Analysis can be seen in the following paragraph).(3)Anatomy and pathology: Under Light Microscope,the gastric mucosa of TG was smoother than that of MG,with smaller punctate hyperemia erosion and inflammatory cells infiltration and without ulcers.(4)Open-field-test,Gastric emptying rate,Ulcer index,5-HT level in serum : all above of TG were inproved(p<0.05);(5)Ulcer inhibition rate was 70.45%.3.i TRAQ detection:(1)Differential protein: Compared with the BG,the number of up-regulated proteins in MG are 53,and the down-regulated are 260.Compared with the MG,the number of up-regulated proteins in the TG are 131,and down-regulated are 26.After crossover,84 number of proteins of post-treatment conversion are obtained(A0a0h2uh9 is rejected as invalid).Ten of them are adjusted upward and then downward,the other in the opposite.(2)GO and KEGG enrichment analysis:Enrichment based on Metascape database :A.Go enrichment is related to 73 biological processes,the most relevant being 13.It mainly involves the catabolism of α-amino acids,the regulation of tricarboxylic acid cycle,the regulation of protein stability,the response of cells to glucose stimulation,the metabolism of hexose,the plasma membrane,the metabolism of monocarboxylic acids,the translation,the metabolism of cellular ketones,RNA splicing with the swollen adenosine Transesterification as nucleophilic reagent,and the positive regulation of receptor-mediated endocytosis,etc.B.KEGG enriched to 20 pathways,the most relevant were 6,including carbon metabolism,2-oxo-carboxylic acid metabolism,splice,valine,leucine and L-Isoleucine degradation,tryptophan metabolism,estrogen signaling pathway and so on.(4)Protein node center network was extracted by analyzing the network interaction of different proteins with STRING database.Conclusion:1.Successfully establish the rat model of CSGU with stagnation of Qi in liver and stomach.2.Ganwei baihe decoction has obvious therapeutic effect on the rat model of CSGU with stagnation of Qi in liver and stomach.3.The therapeutic effect of Ganwei Baihe decoction may be related to the regulation of cellular energy metabolism,oxidative stress,apoptosis,neurotransmitter release,mitochondrial homeostasis,gene alternative splicing,and cytoskeleton-catalyzed inflammatory pathways mediated by actin remodeling are all involved.The relationship between the PI3K/Akt-hpa pathway and CSGU with the stagnation of Qi in liver and stomach was predicted,more experiment to explore.4.The result of WB is consistent with the result of proteomics,which means the result of proteomics is credible. |
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