| Jianpi Yiqi formula is one of the representative prescriptions for CAG treatment based on Traditional Chinese Medicine(TCM),which is condensed from the national"11th five-year plan" support plan.Its clinical curative effect is satisfactory.The previous experimental data show that Jianpi Yiqi formula has inhibitory effect on CAG model rats.Wnt/?-catenin signaling pathway is the canonical Wnt signaling pathway,which is closely related to the occurrence and develop,ment of many diseases.Some studies suggest that this pathway is abnormally activated in chronic gastropathy and may be involved in the pathogenesis of CAG.Based on the results of previous experiments and clinical efficacy,this study explored the effect of Jianpi Yiqi formula on Wnt/?-catenin signaling pathway and CAG model rats.Part one:summary of theoretical researchThis paper summarizes the literature on the pathogenesis,diagnosis and treatment of CAG in modern medicine,and concludes that modern medicine has made some achievements in the aspects of the pathogenesis and pathological evolution of CAG.However,there are still no specific means and methods to improve gastric mucosal atrophy,control or even reverse intestinal metaplasia.In addition,according to the literature of TCM on the etiology,pathogenesis,syndrome differentiation and treatment of CAG,the curative effect of TCM on CAG has certain advantages,spleen and stomach weakness is the pathogenesis basis of CAG.qi stagnation,blood stasis,damp-heat and toxin evil are the pathological products in the process of its occurrence and development;Differentiation of symptoms and signs is the most commonly used treatment of TCM,tonifying spleen and invigorating qi is the basic treatment of CAG,combined with individual treatment is TCM's advantage,which is its flexible treatment.The effect of tonifying spleen and invigorating qi and its prescription on CAG has theoretical basis and certain pharmacological basis.Part two:experimental studyObjective:to observe the effect of Jianpi Yiqi formula on wnt3a,?-catenin,GSK-3? and c-myc protein expression in gastric mucosa of CAG rats,and to explore the preventive and therapeutic effect of Jianpi Yiqi formula on gastric mucosal atrophy and its mechanism.Methods:? modeling:CAG rat model was established by MNNG composite factor modeling method.? grouping and intervention:72 SD male rats were randomly divided into normal group(general feeding),model group(0.9%NaCl),positive drug control group(folic acid 3 mg/kg/d + teprenone 15 mg/kg/d),Jianpi Yiqi recipe low(11 g/kg/d)middle(22 g/kg/d)high dose group(44 g/kg/d),model group 22,the remaining 10 rats in each group,except normal group,the other groups were given drug feed(containing 0.03%ranitidine),using free drinking water(containing 0.17mg/ml MNNG)? specimen collection:after 11 weeks,3 rats in the model group were killed every 2 weeks,the whole stomach was taken out from the stomach,a small piece of gastric tissue was taken at the antrum,body and fundus of the stomach,and the rest tissues were stored in-80 ? refrigerator.Changes ofGSK-3? and c-myc protein expression.? index detection:general observation;He pathological sections were stained and the pathological state of gastric mucosa was observed under light microscope.Western blot was used to detect the expression of wnt3a,?-catenin,GSK-3? and c-myc gene proteins in Wnt/?-catenin signaling pathway.The gray scale analysis of protein bands was performed by image j software.? statistical methods:SPSS 17.0 was used to process the data.Results:? general situation:CAG rats were lighter in weight,less active,more gregarious and less in diet from the second week.? histopathological changes:the model was successful in the 15th week.the pathological findings showed that gastric mucosa became thinner,glands decreased,a large number of gaps appeared,with diffuse or focal inflammatory cell infiltration.intestinal metaplasia was found in some rats' gastric mucosa.at the end of the 19th week,the rats with intestinal metaplasia and dysplasia of gastric mucosa increased.Compared with the model group,both the low,middle and high dose group and the positive drug control group improved the general condition and pathological state of the model rats in different degrees.In contrast,the improvement of general condition and pathological state in low dose group was weaker than that in other intervention groups.The structure of gastric mucosa was clear in positive control group and middle dose group,the epithelial cells were arranged regularly in lower dose group,and there were a small amount of inflammatory cells infiltration in mucosa.The epithelial cells of gastric mucosa in the high dose group were arranged regularly,close to those in the control group.?changes of Wnt/?-catenin signaling pathway protein expression in each group:compared with the blank group,wnt3a,?-catenin and c-myc gene protein expression increased significantly in model group,GSK-3(3 gene protein expression showed low expression or deletion,the difference was statistically significant(p<0.05).B compared with the model group,GSK-3? expression increased,wnt3a and P-catenin decreased in the four groups(p<0.05).C the expression of GSK-3? was significantly increased in the high dose group compared with the other three intervention groups(p<0.05),followed by the middle dose group and the low dose group and the positive drug control group(p<0.05).Wnt3a protein expression was decreased in all the four groups after drug intervention,and dose dependence was observed in Jianpi Yiqi formula,low,middle and high dose groups,there was significant difference between high,middle and low dose groups and positive drug control group(p<0.05).The expression of ?-catenin protein was also decreased in the 4 intervention groups,especially in the high dose group,which was statistically significant compared with the middle and low dose group and the positive drug control group,followed by the middle and low dose group,and its protein expression level was lower than that of the positive drug control group(p<0.05).The expression of c-myc protein was also decreased in the 4 intervention groups,but it was not significantly decreased in the low dose group,and the difference was statistically significant compared with the other 3 groups(p<0.05).Conclusion:Jianpi Yiqi decoction can down-regulate the expression of wnt3a.?-catenin and c-myc protein in gastric tissue during CAG rats,up-regulate GSK-3?protein level,and affect the formation of gastric mucosal atrophy.The results showed that the inhibitory effect of Jianpi Yiqi formula on CAG rats was related to the regulation of Wnt/?-catenin signaling pathway related protein expression by Jianpi Yiqi formula.This study provides a scientific basis for the prevention and treatment of CAG by TCM compound formula. |
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