Protective Effect And Mechanism Of Bilobalide Against Gastric Ulcer In Vivo And In Vitro

Objective:The aim of the study was to reveal the potential efficacy as well as the underlying mechanism of BI(bilobalide)on ethanol/acetic acid-induced lesion in gastric mucosa in vivo/vitro.Methods:Chapter One:Prepared GES-1 cells were treated with BI(10μM,20μM,40μM)for 24h and 400μM H2O2 was added for 1h except the control group.MTT was added to each well to test vitality.Interleukin-6(IL-6),IL-1β,tumor necrosis factor-a(TNF-a)were tested by ELISA in cell supernatant.Western blot technology was adopted to test the expression of P-JNK、JNK、P-Erk、Erk、P-P38、P38、P-IKKα IKKα、P-IKKβ、IKKβ、P-IκBα、IκBα、P-NF-κBp65、NF-κBp65 in cells.Chapter Two:Ethanol(0.2 ml/kg)was applied to induce gastric ulcer in mice model.A total of 50 mice were divided into 5 groups as the control group,ethanol group,OME group(omeprazole pretreated),BI(3mg/kg,6mg/kg)group(bilobalide pretreated).Histopathologic changes in stomach tissues were observed in each group.Superoxide Dismutase(SOD),Maleic Dialdehyde(MDA),TNF-a,IL-6,IL-1β were tested in serum and SOD,MDA,Myeloperoxidase(MPO)in stomach.P-JNK,JNK,P-Erk,Erk,P-P38,P38,P-IKKa,IKKa,P-IKKβ,IKKβ,P-IκBα,IκBα,P-NF-κBp65,NF-κBp65 expression were tested in stomach by Western blot.Chapter Three:Five groups of mice were distributed as the control group,acetic acid group,OME group,BI(3mg/kg,6mg/kg)group with 10 mice in each.Histopathologic changes in stomach tissues were observed in each group.ELISA were adopted to test the content of SOD,MDA,TNF-α,IL-6,IL-1β in serum and SOD,MDA,MPO in stomach.The expression of P-JNK,JNK,P-Erk,Erk,P-P38,P38,P-IκBα,IκBα,P-NF-κBp65,NF-κBp65,LC3-I,LC3-II,LAMP1,mTOR,p-p70S6K,p70S6K,p-4EBP1,4EBP1 were tested in stomach by Western blot.Results:Chapter One:MTT results showed that BI could improve the damage in gastric epithelial cells and decrease the concentration of TNF-a,IL-6,IL-1β with the inhibition of P-JNK,P-Erk,P-P38,P-IKKa,P-IKKp,P-IκBα and P-NF-κBp65 expression.Chapter Two:BI could alleviation the damage in gastric mucosa induced by ethanol and decrease the concentration of MDA,TNF-α,IL-6,IL-1β,increase the concentration of SOD in serum.The concentration of MDA,MPO and P-JNK,P-Erk,P-P38,P-IKKα,P-IKKβ,P-IκBα,P-NF-κBp65 were decreased in gastric mucosa while SOD were increased if pretreated by BI.Chapter Three:BI could alleviation the damage in gastric mucosa induced by acetic acid proved by H-E staining.The concentration of MDA,TNF-a,L-6,IL-1β in serum and MDA,MPO in gastric mucosa were decreased as well as SOD were increased both in serum and in mucosa with the.pretreatment of BI.BI could inhibit P-JNK,P-Erk,P-P38,P-IκBα,P-NF-κBp65,LC3-I,LC3-II,LAMP1,mTOR,p-p70S6K and p-4EBP1 expression in gastric mucosa.Conclusions:BI exerted a gastro-protective effect in vivo and in vitro through the MAPK/NF-κB pathway.In ethanol/acetic acid induced gastric ulcer,BI exerted a gastro-protective effect which was presumed to be associated with MAPK/NF-κB pathway with the inhibition of oxidative stress.Autophagy was tested in acetic acid induced gastric ulcer,and was proved to be involved in the BI protective process.