Effects Of Xiaoyongkuidekang And Its Disassembled Prescriptions On The Expression Of PKA And CREB Proteins In GES-1 Cells Induced By Ethano

Purpose:Exploring the expression and content changes of PKA,CREB protein and mRNA in ethanol induced GES-1 cells treated with the representative formula of "treating carbuncle based on carbuncle"-Xiaoyongkuidekang granules and their functional decoctions,and exploring the mechanism of Xiaoyongkuidekang granules’ functional decoctions in treating peptic ulcers,providing scientific theoretical basis for the evolution of the cause of "toxic heat" in peptic ulcers.Material and method:In the experimental part,the cell model was established.After using CCK-8 to detect and screen the optimal concentration of cell modeling drug and the optimal concentration of drug-containing serum,the rats were randomly divided into blank group,model group,Xiaoyongkuikang group,heat-clearing and detoxicating group,Tuohu Shengji group,Jianpi and Wei group,and Weireqing group with 8 rats in each group.After the cell model was made with the best ethanol concentration,the animal serum containing drugs was intervened for 24 hours,and the material was taken.The expression of PKA and CREB protein in GES-1 cells was detected by Western-blot method;RT-PCR was used to detect the mRNA expression level of PKA and CREB in GES-1 cells.SPSS25.0 was used for statistical analysis.Univariate analysis of variance between groups showed that P<0.05 had statistical difference.Results:1.Western blot method was used to detect the protein expression levels of PKA and CREB in GES-1 cells after treatment with different groups of drug-containing serum.Compared with the model group,the protein expression levels of PKA and CREB in the supernatant of GES-1cells in the Qingre Jiedu group,Tuohu Shengji group,Jianpi Hewei group,Xiaoyongkuikang group were significantly higher than those in the group,with statistical significance(P<0.05).2.RT-PCR method was used to detect the mRNA levels of PKA and CREB genes in GES-1 cells after treatment with different groups of drug-containing serum.Compared with the model group,the mRNA levels of PKA and CREB in GES-1 cells in the Qingre Jiedu group,Tuo Fu Sheng Ji group,Jian Pi He Wei group,and Xiaoyongkuikang group were significantly increased compared to the group,with statistical significance(P<0.05).Conclusion:1.Observing GES-1 cells modeled with ethanol under an optical microscope,it was found that the cell model prepared with 5% ethanol for 3 hours was the best.2.Xiaoyong Kuidekang Granules and their functional decoctions can increase the expression of PKA and CREB proteins and mRNA in ethanol induced GES-1 cells.3.The mRNA and protein content of PKA and CREB in the heat clearing and detoxification group is only inferior to the normal group and the Xiaoyong Kuidekang Quanfang group,and is significantly higher than the other five groups,indicating the existence of the "toxic heat" etiology in peptic ulcers.4.Xiaoyong Kuidekang Granules and their functional decoctions may improve ethanol induced GES-1 cell inflammation by regulating the PKA/CREB signaling pathway,thereby achieving the goal of treating peptic ulcers.