Study On The Protective Effect Of Alpinia Officinarum On Indomethacin-induced Gastric Injury And Its Gastrointestinal Distribution

Objective:To explore the protective effect of Alpinia officinarum on Indomethacin-induced gastric injury and its mechanism,and to explore whether this protective effect is related to the distribution of drugs in the stomach and other digestive organs,so as to provide data support for clinical medication.Methods:By establishing indomethacin-induced gastric injury rat model and gastric mucosal epithelial cell injury model,we explored the protective effect of Alpinia officinarum on indomethacin-induced gastric injury in vivo and in vitro,and studied its mechanism by Western blotting,flow cytometry and other molecular pharmacological methods;A UFLC-MS/MS method was established to investigate the effect of Alpinia officinarum on the distribution of indomethacin and its metabolites in rat stomach and other digestive organs.Results:1.Protective effect of Alpinia officinarum on Indomethacin-induced gastric injury in ratsIn the efficacy study test,compared with the model group(Indomethacin,20mg/kg),the administration group could significantly reduce the rat ulcer index,and the middle dose group of Alpinia officinarum alcohol extract(180 mg/kg)significantly reduced(P<0.001)the rat ulcer index.In Western blotting test,compared with the model group,both the administration group and the positive group could up-regulate the expression level of COX-1 protein,and there was significant difference in the middle dose group of Alpinia officinarum alcohol extract(P<0.05);compared with the model group,both the administration group and the positive group could up-regulate the expression level of COX-2 protein,and there was a very significant difference between the middle dose group and the Alpinia officinarum group(P<0.0001).2.Protective effect of Alpinia officinarum on Indomethacin-induced gastric mucosal epithelial cell injuryIn the efficacy study test,compared with the model group,Alpinia officinarum ethanol extract(0.5,2.5μg/m L)and galangin(0.005,0.05μmol/L)accelerated the proliferation of Indomethacin-induced gastric mucosal epithelial cells in varying degrees,with significant difference(P<0.05).In the mechanism study test,0.05μmol/L galangin and 2.5μg/m L Alpinia officinarum ethanol extract could increase the activity of H~+-K~+-ATPase,increase the expression level of pro-caspase-3 protein and reduce the expression level of cleaved-caspase-3 protein in gastric mucosal epithelial cells,and there were significant differences compared with the model group(P<0.05).3.Effects of Alpinia officinarum on the distribution of Indomethacin and its metabolites in rat stomach and other digestive organsA UFLC-MS/MS method was established for the determination of indomethacin and its metabolites in rat stomach and other digestive organs(stomach,small intestine,large intestine).The retention times of Indomethacin,O-desmethyl indomethacin,N-deschlorodenzoyl indomethacin,Indomethacin Acyl-β-D-glucuronide and Indobufen were 2.51 min,1.88 min,1.53 min,1.76 min and 1.84 min respectively.The specificity,accuracy,precision,stability,matrix effect and extraction rate of the method met the requirements.In the model group,the maximum drug concentration(C-max)of Indomethacin in the stomach was 5065 ng/L,in the small intestine was 2005 ng/L,and in the large intestine was 1065.5 ng/L.In the administration group,the C-max of Indomethacin in the stomach was 8307.50 ng/L,in the small intestine was 1882.5 ng/L,and in the large intestine was 1023 ng/L.After administration,the concentration of ODI in stomach and large intestine decreased significantly,and the concentration of IDAβG in the small intestine decreased from 15825 ng/L to 4856 ng/L.Conclusions:Alpinia officinarum has a protective effect on Indomethacin-induced gastric injury.Its mechanism is related to up-regulating the expression level of COX protein,inhibiting the activation of caspase-3 protein,inhibiting cell apoptosis and gastric acid secretion.In addition,the protective effect of Alpinia officinarum on Indomethacin-induced ulcerative gastric injury was not achieved by reducing the accumulation of Indomethacin in the gastrointestinal tract.It is inferred that the protective effect of Alpinia officinarum is related to the regulation of the activities of transporters and metabolic enzymes in vivo,which still needs to be further studied.