Adlay Seed Protein Dependent On IKK/NF-κB Pathway To Control Inflammation And Improve Insulin Resistance In Type 2 Diabetes Mellitus

Adlayseed(Coix lacryma-jobi L.)is a mature seed of adlay,a gramineous coix genus herbaceous plant that is annual and widely planted in warm regions of Asia,Africa and the Mediterranean coast.Adlay seed addition to the seeds,roots,stems and leaves can be used in our country and has a long history of consumption.Adlay seed is rich in protein resources.In recent years its hypoglycemic effect has caused widespread concern in the community.In this experiment,adlay seed as raw material,alcohol-soluble salting-out method for the preparation of adlay seed protein,water-soluble alcohol precipitation for polysaccharides.Adlay seed proteinin type 2 diabetic mice IKK/NF-κB signaling pathway regulation role,dependent on the signaling pathway regulation of inflammatory cytokines levels and related gene expression were studied,adlay seed protein dependent on IKK/NF-κB signaling pathway regulation of inflammatory cytokines levels and related gene expression in type 2 diabetic mice were studied,and the mechanism of adlay seed protein on the insulin resistance was explored.In this study,the content of adlay seed protein was detected by Kjeldahl method,and the content of polysaccharide was determined by phenol-sulfuric acid method.The extracted protein and polysaccharide content were 67.52% and 68.6% respectively.The model of type 2 diabetes was established by high-fat diet combined with intraperitoneal injection of STZ.After modeling,different doses of adlay seed protein,polysaccharides and their mixture were administered to mice.After gavage for 4 weeks,body weight,fasting blood glucose,organ index,blood lipid,pancreatic tissue morphology,insulin level,insulin sensitivity index and insulin resistance index were measured.Enzyme linked immunosorbent assay(ELISA)was adopted to detect serum levels of tumor necrosis factor-α(TNF-α),interleukin-1(IL-1α,IL-1β),interleukin-6(IL-6)and cyclooxygenase-2(COX-2)content.The method of semi-quantitative PCR(RT-PCR)was used to detect nuclear factor-kappa B inhibitor protein kinase complex(IKKβ),nuclear factor kappa-β(NF-κB)p65,toll-like receptor 4(TLR4),phosphatase and tensin homologue(PTEN),phosphatidylinositol 3-kinase(PI3K),glucose transporter protein 4(GLUT4)and FOXO1 gene expression levels.Animal experiments show that adlay seed protein and polysaccharide gavage can effectively reduce the blood glucose index,visceral index and blood lipid levels in mice,repair pancreatic β-cell and maintain body weight;the results of ELISA showed that adlay seed protein could reduce the levels of many inflammatory cytokines.RT-PCR showed that the expression of IKKβ,NF-κB p65 and TLR4 genes were significantly decreased,and proved adlay seed protein can interfere with IKK/NF-κB signaling pathway,so as to achieve the effect of controlling the secretion of inflammatory cytokines;at the same time,it can inhibit the expression of PTEN,PI3 K,GLUT4 and FOXO1,improve the insulin resistance in type 2 diabetic mice.