Protective Effect And Mechanism Of The Fruit Of Rosa Odorata Sweet Var.gigantea(Coll.et Hemsl.) Rehd.et Wils On Stress-induced Gastric Mucosa Injury

Objective: To explore protective effect of Rosa odorata sweet var.gigantea(Coll.et Hemsl.)Rehd.et Wils on stressful gastric mucosa injury as well as mechanisms of action based on establishing stress-induced gastric mucosa injury model in vitro and in vivo.Methods: Fruits of Rosa odorata sweet var.gigantea(Coll.et Hemsl.)Rehd.Et were analyzed by ultra high performance liquid chromatography(UPLC)-tandem quadrupole(QTOF)-time of flight mass spectrometry(MS)Wils Extract,(FOE)main ingredient.Human gastric epithelial cells(GES-1)were cultured in vitro and treated with different concentrations of FOE(25,50,100 mg/m L)for 24 h,then stimulated with hydrogen peroxide(100 μM)for24 h.CCK8 was used to detect cell viability and observe the effect of FOE on human gastric epithelial cells induced by hydrogen peroxide.The contents of TNF-α,IL-6,IL-1β,MDA and SOD in supernatant were detected by ELISA kit.The protein levels of NF-κB p65,IKKα/β,Keap1,Nrf2 and HO-1 were determined by Western Blot.The migration rate of Nrf2 towards the nucleus and the change of mitochondrial membrane potential were detected by high content screening.In vivo experiment,42 rats were randomly divided into 6 groups: blank control group,model control group,omeprazole(20 mg/kg)group,FOE low-dose(62.5mg/kg)group,FOE medium-dose(125 mg/kg)group,and FOE high-dose(250 mg/kg)group.Ulcer index was evaluated and HE staining was performed on gastric tissue after successful modeling.The contents of inflammatory mediators and oxidative stress factors in serum and gastric tissues were determined.Determination of pepsin activity in gastric juice was carried out.The expression levels of NF-κB p65,IKKα/β,Keap1,Nrf2,HO-1,Bcl2,Bax,Pro-caspase3,Cleaved caspase3,PCNA in gastric tissues were determined by Western blot.The content variation of p-EGFR,p-Src,Cyto-c and cleaved-PARP1 protein were detected by immunohistochemistry,so as to explore conceivable mechanism of action against gastric mucosa injury.Results: A total of 26 compounds were detected from FOE,and 22 compounds were identified by molecular ion peak,secondary mass spectrometry fragment information,UV absorption,retention time and related literature.CCK8 in vitro manifested that FOE could ameliorate hydrogen peroxide-induced injury of GES-1.FOE can significantly reduce the contents of TNF-α,IL-6,IL-1β and MDA in supernatant,whereas increase the content of SOD.FOE decreased the expression of NF-κB p65,IKKα/β and Keap1 proteins,and promoted the expression of Nrf2 and HO-1 proteins.FOE significantly promoted Nrf2 entry towards the nucleus of GES-1 cells and inhibited mitochondrial membrane potential decline.In vivo,FOE could improve the pathological injury of gastric tissue induced by water immersion and restraint stress compared with the model group.FOE significantly decreased the contents of MDA,i NOS,TNF-α,IL-6,IL-1β,GAS and ET in serum and gastric tissues,and increased the contents of SOD,CAT,GSH-Px,IL-10,PGE2,TGF-α and SS.FOE could lower pepsin activity in gastric juice.FOE suppressed NF-κB p65,IKKα/β,Keap1,Bax,Cyto-c,pro-caspase3,Cleaved-caspase3,cleaved-PARP1 protein expression in gastric tissues,and promoted the level of Nrf2,HO-1,Bcl2,p-EGFR,p-Src and PCNA proteins.Conclusion: FOE has a protective effect on hydrogen peroxide-induced injury of GES-1,the mechanism of which may be inhibiting inflammatory response and oxidative stress.FOE may play a protective role against stress-induced gastric mucosal damage in rats by inhibiting inflammation,oxidative stress and EGF-induced mitochondrial apoptotic pathway apoptosis pathways.