Analysis Of The Medication Rule Of Chronic Atrophic Gastritis And The Preliminary Mechanism Of The Core Components’ Prevention To Gastritis

Objective: To summarize the medication rules and core Chinese medicine(CCM)of the treatment of chronic atrophic gastritis(CAG)with spleen-stomach weakness(SSW),and to preliminarily explore the mechanism of treating CAG with SSW with CCM,and to further verify the prediction mechanism through in vitro cell experiments.Methods: Use the Chinese medicine treatment of CAG with SSW that published in databases such as CNKI from January 1979 to November 2022,and the clinical observation,expert experience or retrospective medical record analysis and other journals and literature,which compliance with the inclusion criteria,to establish a database.Use the Ancient and Modern Medical Case Cloud Platform v2.3.5 to sort out and analyze the basic information of the literature,and summarize the medication rules and CCM.Select the active ingredients and targets of CCM in the TCMSP database by network pharmacology method,retrieve CAG-related targets in the disease database such as Gene Cards,and screen the common target genes of drugs and diseases,so as to construct the analysis gene interaction network.The active ingredients corresponding to the common target genes of drugs and diseases were screened to build a "active ingredient-target-disease" network map of CCM,and the GO function analysis and KEGG pathway enrichment analysis were performed for common target genes to speculate the key mechanism of its action on CAG with SSW.After 2-hour freeze-dried powder with Sijunzi Decoction,N-Methyl-N’-nitro-N-nitrosoguanidine(MNNG)was used to construct an in vitro model of GES-1 cell CAG.After 24 h,CCK-8 was used to detect cell proliferation,Hochest33258 was used to detect apoptosis,RT-q PCR was used to detect the expression levels of FOS,NR3C1,MAPk1,and IL-6,Western blot was used to analyze the expression of apoptosis-related proteins(Bad,Bcl-2,and Cleaved caspase-3),and Western blot was used to detect the expression changes of molecules STAT1,STAT3,ERK1/2,and phosphorylated proteins.Results:1.This study selected 80 literatures on the treatment of CAG with insufficiency of the spleen,including a total of 80 prescriptions.Among the included prescriptions,the frequency of glycyrrhiza were the highest,followed by Codonopsis and Rhizome atractyloides.2.According to the statistics of TCM with frequency higher than 5,the top 4 most frequencyly used medicines were: deficiency-nourishing drugs,activating medicinal materials,inhibiting-damp drugs,promoting-circulation drug,among which deficiency-nourishing drugs had the highest frequency,including qi-invigorating herbs,blood tonic and Yang tonic.3.The main channel-tropism of medicine werr spleen meridian,stomach meridian,followed by lung channel,heart meridian,liver meridian.The four natures of medicine were mainly warm drugs and mild medicines while warm drugs were the most,which followed by mild medicines,mild cold drugs,mild warm drugs,and no major cold drugs.The five flavors were mainly sweet and hot.4.The CCM for the treatment of CAG with insufficiency of the spleen were codonopsis pilosula,poria cocos,rhizoma atractylodis macrocephalae and liquorice.5.The CCM contained acetoxyatractylodes,8β-ethoxy-atractylodes-II,spinachsterol,ligustrindole,stigmasterol,7-methoxy-2-methyl isoflavone,ivy saponin,licorice alcohol and other active ingredients.The core targets of CAG with insufficiency of the spleen were STAT3,MAPk1,STAT1,FOS,IL6,HSP90AA1 and NR3C1.6.Through GO analysis,it is concluded that CCM affected molecular functions like protease binding,phosphatase binding,protein phosphatase binding in membrane raft,membrane microdomain and membrane region to participate in the cell response to lipopolysaccharide,oxidative stress response,reactive oxygen metabolism process and other biological processes to treat CAG with insufficiency of the spleen.IL-17 signaling pathway,TNF signaling pathway and Toll-like receptor signaling may be the main signal transduction pathways for the treatment of CAG with insufficiency of the spleen that were predicted by KEGG thoroughfare analysis.7.The cytotoxicity test showed that Decoction of Four Noble Drugs could significantly inhibit the proliferation of GES-1 cells and produce significant toxicity when the concentration reached 2mg/m L(P<0.01).The toxic effects of MNNG on GES-1 cells were significantly inhibited by Decoction of Four Noble Drugs lyze-dried powder of 0.1mg/m L,0.5mg/m L and 1mg/m L respectively(P<0.01).The results of staining at Hochest33258 showed that the nucleus of gastritis group was hyperpyretic and fragmented.Different concentrations of Decoction of Four Noble Drugs freeze-dried powder could significantly inhibit the nuclear damage in gastritis group.Compared with Control group,the expressions of FOS,NR3C1,MAPk1 and IL-6 in gastritis cell model were significantly increased(P<0.01).However,different concentrations of Decoction of Four Noble Drugs could significantly inhibit FOS,NR3C1,MAPk1,and IL-6(P<0.01).8.In western blot assay,MNNG significantly increased Bad and Cleaved caspase-3expression(P<0.01)and inhibited Bcl-2 expression(P<0.01)after intervention in GES-1cells compared with Control group.Decoction of Four Noble Drugs at different concentrations significantly inhibited the expression of Bad and Cleaved Caspase-3(P<0.01)and promoted the expression of Bcl-2(P<0.01).Compared with the Control group,MNNG could significantly promote the phosphorylation levels of STAT1,STAT3 and ERK1/2 after intervention in GES-1 cells(P<0.01).Decoction of Four Noble Drugs at different concentrations could significantly inhibit the phosphorylation levels of STAT1,STAT3 and ERK1/2(P<0.01).Conclusion: In the prescriptions for the treatment of CAG with insufficiency of the spleen involved in the research,the most common drugs were deficiency-nourishing drugs while the main effects were strengthening the spleen and replenishing qi,reducing the inverse and removing ruffians,removing blood stasis and relieving pain.The core Chinese medicines were Codonopsis codonopsis,poria codonopsis,atractylodes and glycyrrhiza with the key active ingredients of acetoxyatractylodes ketone,8β-ethoxy-atractylodes lactone-II,spinachsterol,ligustrindole,stigmasterol,7-methoxy-2-methyl isoflavone,ivy saponin,glycyrrhiza alcohol and other substances.It mainly acted on key targets such as STAT1,FOS,and IL6.It may regulate Bad,Cleaved caspase-3 and related protein expression,and phosphorylation levels of STAT1,STAT3,and ERK1/2 through IL-17 signaling pathway,TNF signaling pathway,to inhibit GES-1 cell proliferation,induce GES-1 apoptosis,and achieving the effect of the treatment of CAG with insufficiency of the spleen.