| Objective:1.To explore the changes of the material basis and the treatment of chronic atrophic gastritis of Ginseng Radix before and after combined with Atractylodis Macrocephalae Rhizoma.2.To study the mechanism of Ginseng Radix and Atractylodis Macrocephalae Rhizoma on the treatment of chronic atrophic gastritis by regulating aquaporin,oral and intestinal flora,and provide evidence for clinical rational drug use.Method:1.The ginsenoside of GR before and after combined with AMR were extracted with 60% ethanol,ginsenoside were enriched and purified by macroporous resin.The changes of ginsenoside of GR before and after combined with AMR changes was studied by HPLC.2.The volatile oils of AMR before and after combined with GR were extracted by steam distillation.The contents of volatile oil was analyzed by GC-MS.3.Using MNNG to replicate the model of chronic atrophic gastritis of spleen deficiency,After 70 days of modeling,the corresponding drug was given to the drug-administered group by the amount of the crude drug,and the drug was administered once a day for 60 consecutive days.The microscopic and ultrastructural changes of gastric mucosa in each group were observed by HE staining and scanning electron microscopy.The expression of AQP3 and AQP4 protein in gastric mucosa of rats with chronic atrophic gastritis was detected by immunohistochemistry.The expression of AQP3 and AQP4 mRNA expression was detected by PCR.Through high-throughput sequencing technology,the total DNA of intestinal and oral microorganisms was extracted,and the changes of intestinal and oral flora were detected and correlated analysis was performed by using SPSS software.Results:1.The results showed that the content of Rg1,Re,Rf,Rb1,Rc,Rb2 and Rd inginsenoside increased when GR combined with AMR,and new substances were produced.2.The GC-MS method was used to determine the volatile oil.18 compounds and24 compounds were identified from the volatile oil of AMR and AMR combined with GR.The volatile oils identified 24 compounds.The highest relative volatiles in the two groups were atractylone,was 45.94% and 51.67%.In addition,the most abundant volatile oil components and the most abundant species were olefinic compounds.3.Compared with the normal group,the gastric tissue pathological score of the model group was significantly increased(P<0.05),compared with the model group,the pathological scores of the treatment groups were significantly lower than those of the model group(P<0.05),the pathological score of gastric tissue was decreaser in each compatibility group than no compatibility group.Electron microscopy showed that the gastric mucosa of the model group showed wrinkling and bulging,deformation of the gastric pit,and atrophy of the epithelial cells of the small concave wall.Compared with the model group,the size of the gastric pits tends to be regular.Compared with the normal group,the expression of AQP 3 protein and AQP 4 protein in the gastric tissue sections of the model group were significantly decreased(P<0.01).Compared with the model group,the expression of AQP 3 protein and AQP 4 protein in the gastric tissue sections of the model group were increased Compared with the blank group,the expression of AQP 3 mRNA in the gastric mucosa of the model group was significantly increased.Compared with the model group,the expression of AQP 3 mRNA in the gastric mucosa of each group was decreased.Compared with the blank group,the expression of AQP 4 mRNA in the model group was significantly decreased(P<0.05).Compared with the model group,the expression of AQP 4 mRNA in the gastric mucosa of each group was increased.4.Sequence analysis of 16 S rDNA gene showed that the diversity and abundance of intestinal and oral flora in the treatment group were significantly increased(P<0.05),in administration group the number of pathogenic bacteria in the intestines and oral cavity were reduced,and the imbalance of intestinal and oral flora were improved.Correlation analysis showed that the pathological inflammatory score of rats wascorrelated with the abundance and diversity of oral and intestinal flora and the differential bacteria at the level of phylum and genus(P<0.05).Conclusions:1.The content of Rg1,Re,Rf,Rb1,Rc,Rb2 and Rd in ginsenoside increased when GR combined with AMR,and new substances were produced,this may be the material basis for the increased efficacy of GR combined with AMR for chronic atrophic gastritis.2.The volatile oil component changed when GR combined with AMR,the content of atractylone in volatile oil decreased after compatibility,which reduced the dryness.In addition,new substances were produced,and the olefinic compounds were changed,this may be the material basis for the increased efficacy of GR combined with AMR for chronic atrophic gastritis.3.The effective parts of GR and AMR could alleviate the inflammation of gastric mucosa in rats with chronic atrophic gastritis,increased the expression of AQP 3protein and AQP 4 protein,made the expression of AQP 3 and AQP 4 mRNA tend to normal,and improved the water and salt metabolism of gastric mucosal cells.It relieved the atrophy of the gastric mucosa and restored the physiological function of the stomach to normal.4.The effect of polysaccharides extracted from GR combined with AMR on chronic atrophic gastritis was better than that of GR polysaccharide,AMR polysaccharide and GR polysaccharide + AMR polysaccharide.5.The therapeutic effect of GR combined with AMR group was better than that of single-drum combination group,which was the best in all drug-administered groups,suggesting that GR combined with AMR can enhance the treatment of chronic atrophic gastritis.6.The effective parts of GR and AMR could reverse the abundance and diversity of oral and intestinal flora to a certain extent,reduced the pathogenic bacteria in the intestines and oral cavity,it is suggested that the regulation of GR and AMR on oral and intestinal microecology is one of the mechanisms for the treatment of chronicatrophic gastritis. |
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