| Objective:To observe the effect of Jianpi Huayu Jiedu Recipe on histopathological changes of gastric mucosa in rats with chronic atrophic gastritis,CD34+pathological microangiogenesis,Notch-1 and DLL-4 protein expression in gastric mucosal epithelium,and to explore the possible mechanism of Jianpi Huayu Jiedu recipe in treating chronic atrophic gastritis.Methods:1.Fifty-two male SD rats of SPF grade were randomly divided into blank group(10 rats)and model group(42 rats).Four rats died during the establishment of the model.At the end of the eighth week,two rats were randomly selected to take gastric mucosa tissue for histopathological examination to verify the establishment of the model,Then the model group was divided into model group(9 rats),folic acid group(9 rats),Jianpi Huayu Jiedu recipe high-dose group(9 rats).The low dose group of Jianpi Huayu Jiedu Formula(9 rats).Each group was abbreviated as blank group,model group,folic acid group,high-dose group,low-dose group.2.The CAG rat model was reproduced by"MNNG-ethanol" compound method in the model group.The model was made for 20 weeks continuously,and the treatment was started by intragastric administration at the 9th week.Rats in the blank group and model group were given saline at the dose of 10 ml/kg/d,rats in the folic acid group were given 1.8 mg/kg/d,rats in the high-dose group and low-dose group were given 15.8 g/kg/d and 7.9 g/kg/d respectively for 12 weeks.During the period,the dosage was adjusted according to the weight change of rats.3.At the end of the experiment,three pieces of gastric mucosal epithelial tissue were taken from the gastric body-antrum junction of rats.The pathological changes were observed under light microscopy after HE staining.The microvessel density of CD34+ and the expression of Notch-1 and DLL-4 in gastric mucosal epithelial tissue were detected by immunohistochemistry.SPSS software was used to analyze the statistical data,and the results were expressed as mean(+standard deviation).Result:1.General changes:The rats in the model group had loose stool,reduced food intake,lower average body weight than the blank group,withered fur,depression,reduced activity and so on.The above-mentioned situation of rats in each treatment group has been improved to varying degrees,especially in the high-dose group and low-dose group.2.Histopathological changes:HE staining showed that the gastric mucosal proper glands atrophied obviously,the arrangement of glandular ducts was distorted,the nuclei were pyknosis,the ratio of nucleus to plasma was imbalance,and intestinal epithelial metaplasia and dysplasia appeared.After treatment,the number of gastric mucosal proper glands increased,the arrangement of glands was regular,intestinal metaplasia and dysplasia were significantly improved in the high-dose group and low-dose group.The rats in folic acid group also improved,but the degree was less than the high-dose group and the low-dose group.3.Changes of CD34+microvessel density:Compared with the blank group,the CD34+microvessel density of gastric mucosa in the model group increased significantly(P<0.01).Compared with the model group,the microvessel density of CD34+in folic acid group decreased,but the difference was not statistically significant(P>0.05).The microvessel density of CD34+ in gastric mucosa of rats in high-dose group and low-dose group decreased significantly(P<0.01).Compared with folic acid group,the mierovessel density of CD34+in gastric mucosa of rats in high-dose group and low-dose group decreased more significantly(P<0.01).There was significant difference(P<0.01),and the decrease of CD34+microvessel density in gastric mucosa of rats in high-dose group was more significant than that of rats in the low-dose group(P<0.05).4.Changes of Notch-1 and DLL-4 protein expression:Notch-1 expression was n egative in gastric mucosal epithelium of rats in each group.Compared with the blank group,the expression of DLL-4 protein in gastric mucosa of the model group was significantly increased(P<0.01).Compared with the model group,the expression level of DLL-4 in gastric mucosa of rats in folic acid group decreased,but the difference was not statistically significant(P>0.05);the expression level of DLL-4 in gastric mucosa of rats in the high-dose group and low-dose group decreased significantly(P<0.01);compared with the folic acid group,the expression level of DLL-4 in rats in high-dose group decreased more significantly(P<0.01).The difference was statistically significant(P<0.05).The expression of DLL-4 protein in the low-dose group decreased slightly,but there was no significant difference(P>0.05).Conclusion:1.Jianpi Huayu Jiedu Decoction can improve and reverse the pathological changes of gastric mucosa in CAG rats,which may be related to the reduction of CD34+pathological microvessel density in gastric mucosal epithelial tissue and the inhibition of DLL-4 protein expression by Jianpi Huayu Jiedu Decoction.2.Notch-1 in gastric mucosal epithelial tissue of rats in each group showed no significant difference,suggesting that Notch-1 may not be a positive marker and therapeutic target for CAG lesions. |
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