Screening Of Natural Antioxidant Compounds To Reduce The Toxicty Of Gluten And Their Molecular Mechanisms

The incidence of celiac disease has risen to 1-8% in industrialized countries and caucasians.In our country,the iceberg of celiac disease is also floating.In the Shandong population,which is dominated by wheat,the incidence of celiac disease reached 0.76%.Currently,strict adherence to a gluten-free diet for the rest of life is the best way to treat celiac disease.However,because China's gluten-free food production chain is still not perfect and people's awareness of gluten-free diet is lackness,and wheat flour and gluten protein are often used as ingredients in food processing,so it is difficult to completely avoid eating gluten in daily life for celiac patients.And the long-term gluten-free diet will affect the social activities of patients,which will put a burden on their psychology and economy.In this study,caco-2 cells were used to construct the oxidative stress model of celiac disease,and modern molecular biology technology was applied to screen natural antioxidants that can reduce the toxicty of gluten and study their molecular mechanisms.The main methods,results and conclusions are as follows:1.Establishment of oxidative stress model of celiac diseaseCaco-2 cells were treated with gluten-derived peptide p31-43 at different concentrations and for different periods of time,and the intracellular ROS level,SOD enzyme activity,MDA and GSH contents were used as the detection indexes to successfully construct the oxidative stress model of CD based on caco-2 cells.2.Screening of natural antioxidants to reduce the toxicty of glutenChoose different concentrations and different kinds of natural antioxidants pretreatment the cell model which has been successfully established.And then screening the natural antioxidants which can reduce the toxicty ofgluten proteinfrom oxidative damage,celiac disease markers and inflammatory factor expression and intestinal epithelial barrier dysfunction induced by p31-43.Finally,quercetin,epigallocatechin gallate,genistein,?-carotene,myricetinkaempferol and resveratrol were successfully screened from 7 natural antioxidants.3.Study on molecular mechanisms of antioxidants reducing the toxicty of glutenThe effects of antioxidants on the expression of key genes in Nrf2,mTOR and SIRT signaling pathways were studied by Q-PCR technology,so as to conduct a preliminary study on its molecular mechanism.The results showed that the three natural antioxidant had different action mechanisms,among which the treatment of resveratrol can promote the expression of sirt1 and sirt3 genes,indicating that the mechanism of resveratrol reducing the toxictyof gluten protein is related to the activation of SIRT signaling pathway.