Effects Of Jianpi Yiqi Formula On Expression Of Notch Signaling Pathway Related Proteins In Gastric Tissue Of Rats With CAG

OBJECTIVE:To study the effect of Jianpi Yiqi formula on the changes of gastrin(GAS),somatostatin(SS)and prostaglandinE2(PGE2)content in serum,gastric pathological changes and Notch signaling pathway related protein in CAG model rats,so as to explore the mechanism of Jianpi Yiqi formula on treating CAG.METHODS:10 of 65 SD rats were set as blank group.SDCAG rats were established by using the MNNG,ranitidine and disorder diet.Then 50 SDCAG were randomly divided into model group(0.09%sodium chloride solution),positive drug group(folic acid 2.7mg/kg/D+teprenone 13.5mg/kg/D),Jianpi Yiqi formula low-dose group(13.1g/kg/D),Jianpi Yiqi formula midium-dose group(26.2g/kg/D)and Jianpi Yiqi formula high-dose group(52.4g/kg/D).10 rats in each group.Each rat were given 1ml/100g solution by gavage for 8 weeks.Blank group were given 1ml/100g 0.09%sodium chloride solution by gavage for 8 weeks,too.The dosage was calculated according to Pharmacology Experimental Methodology,The state and body weight of rats in each group were recorded.The gastric tissues were stained with hematoxylin-eosin staining(HE).The contents of GAS,SS and PGE2 in serum were detected by Enzyme linked immunosorbent assay(ELISA),and the expression of Ki67 and Notch signaling pathway related proteins were detected by immunohistochemistry(IHC)and Western Blot(WB).RESULTS:①General conditions:The rats in the blank group had nice hair and physical state,normal diet and body weight.The rats in the model group had bad hair,which was gloomy and easy to fall off.The spirit of CAG rats was dispirited,the activity was reduced,the daily feed and drinking water were surplus,and the body weight was significantly lower than that in the blank group(P<0.01).The hair,mental state,activity,diet and body weight of the intervention groups were close to the blank group.Among all the intervention groups,the Chinese medicine intervention groups were better.② Histopathological morphology:In the blank group,the epithelial structure of gastric mucosa was clear,the glands were arranged orderly.In the model group,the epithelium of gastric mucosa arranged disorderly.In each intervention group,the gastric mucosa epithelium got thickened,the glands increased,the arrangement tended to be neat,the inflammatory cell infiltration decreased,and the intestinal metaplasia improved.The medium dose Jianpi Yiqi formula group was the best,and it was similar to the blank group.Less glands disappeared,the arrangement was regular,and there was no obvious mucosal atrophy and intestinal metaplasia.③Serum indexes:Compared with the blank group,the content of GAS,SS and PGE2 in the model group were significantly lower(P<0.01).Compared with the model group,the content of SS and PGE2 in the positive drug group were increased significantly(P<0.05,P<0.01),the content of SS and PGE2 in the low-dose Jianpi Yiqi formula group were increased(P<0.05),the content of SS and PGE2 in the medium-dose group were increased significantly(P<0.01),the content of GAS in the high-dose group was increased(P<0.05),and the content of SS and PGE2 were increased more significantly(P<0.01),but there was no significant difference among the low,medium and high-dose groups(P>0.05).⑤ Ki67 expression:The Ki67 expression in the model group was higher than that in the blank group(P<0.05).Compared with the model group,the Ki67 expression in the positive drug group and Jianpi Yiqi formula groups were increased in varying degrees,the Ki67 expression in the low-dose group was the most obvious(P<0.01),and there was no significant difference between the Chinese medicine groups(P>0.05).⑤ Protein expression of Notch signaling pathway:Compared with blank group,Notch 1,Notch2,Jagged2,DLL1,Hes1 and CyclinD1 in model group were decreased(P<0.05,P<0.01).The results of IHC and WB in the intervention group were little different.Compared with the model group,the results of IHC showed the expression of Notchl in the positive drug group was significantly promoted(P<0.01),while WB results showed the up-regulation of Nothl,Notch2 and Hes1.The results of IHC showed the expression of Notch1,Notch2 and CyclinD1 were up-regulated in low-dose group(P<0.05,P<0.01),while WB results showed the up-regulation of all the test index(P<0.05,P<0.01).The results of IHC showed Notch1,Notch2,Jagged2,DLL1 and CyclinD1 in middle-dose group(P<0.05,P<0.01),while WB results showed the up-regulation of Notchl,Notch2,DLL1 and Hes1.IHC results showed Notch 1,Notch2,Jagged2 and Hesl were increased in high-dose group(P<0.05,P<0.01),while WB results reduced the expression of Notchl,Notch2 and Hes1.There was no significant difference among low,medium and high-dose groups(P>0.05).CONCLUSION:Jianpi Yiqi formula can improve the physical condition and gastric mucosal atrophy of CAG rats,up-regulate the content of serum GAS,SS,PGE2,up-regulate Ki67 gene and the expression of Notch signaling pathway related proteins,so as to promote the proliferation of gastric mucosal cells and improve the atrophy.We believe that the inhibitory effect of Jianpi Yiqi formula on CAG rats is related to the abnormal expression of Notch signaling pathway related proteins.This result explains part of the mechanism of therapeutic effect of Jianpi Yiqi formula on CAG to a certain extent.As for how to regulate the micro targets of signaling pathway proteins,further research is needed.