The Differential Expression Of Target Genes In Gastric Ulcer Rats With Syndrome Of Liver Stagnation And Spleen Deficiency And Traditional Chinese Medicine Intervention Study

Objective By observing the influence of traditional Chinese medicine(TCM)on the differential expression of the target gene in gastric ulcer rats with syndrome of liver stagnation and spleen deficiency,to discuss the functional mechanism of TCM’s intervention on gastric ulcer with syndrome of liver stagnation and spleen deficiency,and to provide more favorable experimental basis for clinical treatment.Methods ①By applying the immortalized primary cells line technique,genetic vectors of the target genes in the model of gastric ulcer with syndrome of liver stagnation and spleen deficiency were constructed by the lentiviral vector and transfected into those gastric cells of a normal fetal rat.②By using MTS colorimetric method in the cells growth experiments and Laser Scanning Confocal Microscope,the differential expression of gene M、I、G、F had been observed and verified.③After observing the intervention towards the expression of target gene by Chaihuang Weikuining,the curative effect of this experiential formula had been confirmed and its functional mechanism found.Result:①Primary gastric epithelial cell line was successfully set up and transfected by applying the technique of lentiviral vector.②According to the results of MTS colorimetric method in the cells growth experiments,the transfection was perfect in the certain density(0.5μg/μ1-1μg/μl and at the certain time(24h after the addition).The cells was growing fast after the transfection of gene M、F、G and became slow after the transfection of gene I.After the intervention of TCM,the cells transfected with gene M、F、G were growing faster(P<0.05 或 P<0.01),among of which,the cells transfected with gene M were growing the fastest by the intervention of Huangqi Jianzhongtang,and the cells transfected with gene F、G were growing the fastest by the intervention of Chaihuang Huikuining.After the intervention of TCM,the cells transfected with gene I were growing slower(P<0.05),and the slowest by the intervention of the high dose Chaihuang Huikuining.③ The observation of Laser Scanning Confocal Microscope showed that the pictures of those cells transfected with gene M、F、G were more bright than before the transfection.And the brightness and the area of transfected with gene M、F、G were gradually added after the intervention of TCM.And the cells transfected with gene F、G were the brightest and the largest by the intervention of the middle dose Chaihuang Wuikuining,while the cells transfected with gene M by the intervention of Huangqi Jianzhongtang.The observation of Laser Scanning Confocal Microscope also showed that the pictures of those cells transfected with gene I were less bright than before,which was more obvious after the intervention by TCM and the most obvious by the intervention of the middle dose Chaihuang Huikuining.Conclusion ①The technique of lentiviral vector can be successfully applied into the construction of target gene vectors in gastric ulcer with syndrome of liver stagnation and spleen deficiency.② The gastric ulcer with syndrome of liver stagnation and spleen deficiency is closely related with the differential expression of gene M、I、F、G in the gastric mucosa.③ The function of Chaihuang Weikuining treating gastric ulcer with syndrome of liver stagnation and spleen deficiency might be worked by two-way regulation(up/down)of differential expression of some genes.