Study On The Mechanism Of Norartocarpetin Against Acute Gastritis Based On Network Pharmacology Strategy

Acute gastritis(AG)is an acute gastric mucosal inflammation caused by various pathogenic factors(ethanol,infection,eating stimulating food,severe trauma,massive bleeding,surgery,shock,extreme mental tension,etc.).According to statistics,gastric hemorrhage caused by acute erosive hemorrhagic gastritis accounts for 10%～30% of all cases of upper gastrointestinal bleeding.With the rapid development of economy and society,the incidence rate of AG has increased significantly,which has seriously affected people’s quality of life.The western medicine treatment of AG has certain side effects and limitations.Modern pharmacological studies have proved that traditional Chinese medicine has anti-inflammatory effect with less toxic and side effects.Therefore,finding safer and more effective anti Ag drugs from natural products has become the trend of pharmaceutical research.In the early stage,the research group isolated the flavonoid chemical monomer norartocarpetin(NTP)from the root of Cinnamomum bungeanum,a plant belonging to the genus potomic in mulberry family.Previous studies have shown that NTP has anti-inflammatory activity.In view of this,this topic first evaluates the potential of NTP in the treatment of Ag based on the network pharmacological method and predicts the relevant key targets,then determines the safety range of NTP through oral acute toxicity test,and finally verifies its anti Ag efficacy by animal pharmacological model,and discusses the relevant mechanism.Objective: To evaluate the effect of NTP on aspirin hydrochloric acid induced acute gastritis in mice and explore its related mechanism,so as to lay an experimental basis for the further development and utilization of NTP.Methods:(1)Network pharmacology was used to evaluate the potential of NTP against acute gastritis and screen the target.(1)Enter NTP into Pub Chem website and search its chemical structure.(2)Search NTP compound targets through Pharmmapper website and Swissadme website,merge the search results and delete duplicate compound targets.(3)Search the disease targets of acute gastritis through Disgenet website,Drugbank website,Genecards website and OMIM website,merge the search results and delete the duplicate Ag disease targets.(4)Using Venny website to find the intersection target of compound target and disease target,input the intersection target into string website for protein interaction,and screen the key target from the intersection target with cyto scape software.(2)Acute toxicity evaluation of NTP:(1)thirty SD mice were randomly divided into blank group,solvent group and drug group,with 12 mice in each group,6 males and 6 females respectively.(2)The acute toxicity grading method was used.With the initial dose of 2000 mg / kg,NTP(dissolved in 5%CMC Na)was administered orally to mice in the treatment group,and the gavage volume was 40 ml / kg.(3)After administration,observe continuously for 14 days.After isoflurane anesthesia,take blood through the orbit,centrifuge,prepare serum,and store it in a 4 ℃ refrigerator for standby.The mice were killed after cervical spondylectomy,and the heart,liver,spleen,lung and kidney were taken,weighed and the organ index was calculated;The liver and kidney tissues were fixed with 4%paraformaldehyde solution for 48 hours and then stained with he.(4)The levels of serum alanine aminotransferase(ALT),glutamate aminotransferase(AST),alkaline phosphatase(ALP),serum total protein(TP),serum albumin(ALB)and blood urea nitrogen(BUN)were measured to evaluate the acute hepatotoxicity and nephrotoxicity of NTP.(3)Efficacy verification and mechanism study of NTP against acute gastritis:(1)60 male SD mice were randomly divided into control group,model group,positive drug group,NTP low dose group,NTP medium dose group and NTP high dose group,with 10 mice in each group;Except the blank group,other mice were induced by 2% aspirin-1.84% hydrochloric acid(V/V,1:1).(2)One hour after modeling,mice in low,medium and high dose groups were given different doses of NTP(25,50 and 100 mg / kg)by gavage;Mice in the positive drug group were given Sanjiu Weitai(5196 mg / kg);Mice in blank group and model group were gavaged with the same volume of 0.5% CMC Na solution;The drug was administered 4 times with an interval of 4 hours.(3)5 hours after the last administration,isoflurane anesthesia,blood was taken through the orbit,centrifuged,and serum was prepared,which was stored in a 4 ℃ refrigerator for standby.;The mice were killed after cervical spondylectomy,and the gastric tissue was taken out and fixed with 4%paraformaldehyde solution.(4)Determination of interleukin-1 in mouse serum by ELISA β(IL-1 β)、 The level of nitric oxide(no)in gastric tissue was detected by microplate method;Pathological sections of stomach were stained with he;Cyclooxygenase-2(COX-2)and nuclear factor kappa-B(NF)in gastric tissue were measured by Western blot-κ B)Protein expression of.Results:(1)Screening of potential key targets of NTP in the treatment of acute gastritis:(1)255 compound targets of NTP were searched through pharmmapper and swissadmi websites.(2)1337 acute gastritis disease targets were searched through websites such as disgenet,drugbank,genecards and OMIM;(3)The compound target and disease target were put into Venny website,and a total of 104 intersection targets were screened.The intersection targets were put into string website for protein interaction.39 potential key targets were screened from the intersection targets with cyto scape software,including prostaglandin endoperoxidase synthase 2(PTGS2),which is also known as cyclooxygenase-2(COX-2).(2)Acute toxicity evaluation of NTP: compared with the control group,the mice in NTP administration group had no death and obvious poisoning during the observation period,and there was no significant difference in weight growth trend and organ index;There was no significant change in serum ALT,AST,ALP,TP and bun(P > 0.05).The alb level of some mice in male vehicle group,male drug group and female vehicle group decreased,but the difference was not statistically significant(P > 0.05).(3)Efficacy verification and mechanism of NTP against acute gastritis: all dose groups of NTP can significantly reduce IL-1 in mice β And no levels(P < 0.01 or P < 0.05)in a dose-dependent manner;The pathological section of gastric tissue showed that the gastric mucosal epithelium in the model group was missing and necrotic,accompanied by bleeding,edema and inflammatory cell infiltration.In the administration group,the inflammatory infiltration of gastric tissue was reduced and the gastric injury was improved;The gastric tissue of mice in the administration group significantly down regulated COX-2 and NF-κ B protein expression.Conclusion:(1)The results of network pharmacology suggest the potential of NTP as an anti Ag drug,and predict its potential target;(2)The acute toxicity test showed that the median lethal dose(LD50)of NTP was 4000-5000 mg / kg,which was a low toxicity drug;(3)The effect of aspirin hydrochloride on the induction of Conag in mice was verified-κ B expression,reduce serum IL-1β、 It is related to the content of NO in tissues.This study laid the experimental data for the further study of anti acute gastritis drugs based on NTP.