Abstract
BackgroundPancreatic islets are known to contain low level of antioxidants that renders them vulnerable to oxidative stress. Nrf2 is the master regulator of numerous genes, encoding antioxidant, detoxifying, and cytoprotective molecules. Activation of Nrf2 pathway induces up-regulation of numerous genes encoding antioxidant and phase II detoxifying enzymes and related proteins. However, little is known regarding the role of this pathway in human islet cells. The aim was to investigate the effect of Nrf2 activator (dh404, CDDO-9,11-dihydro-trifluoroethyl amide) on human islet cells.MethodsHuman islets were obtained from cadaveric donors. After dh404 treatment, Nrf2 translocation, mRNA expression, and protein abundance of its key target gene products were examined. The proportion of dh404-treated or non-treated viable islet beta cells was analyzed using flowcytemetry. The cytoprotective effects against oxidative stress and production of inflammatory mediators, and in vivo islet function after transplantation were determined.ResultsNrf2 nuclear translocation was confirmed by con-focal microscope within 2 hours after treatment, which was associated with a dose-dependent increase in mRNA expression of anti-oxidants, including NQO1, HO-1, and GCLC. Enhanced HO-1 expression in dh404 treated islets was confirmed by Western Blot assay. Islet function after transplantation (2000 IEQ/mouse) to diabetic nude mice was not affected with or without dh404 treatment. After induction of oxidative stress with hydrogen peroxide (200 μM) the proportion of dh404-treated viable islet cells was significantly higher in the dh404-treated than untreated islets (74% vs.57%; P<0.05). Dh404 significantly decreased production of cytokines/chemokines including IL-1β, IL-6, IFN-γ and MCP-1.ConclusionTreatment of human pancreatic islets with the potent synthetic Nrf2 activator, dh404, significantly increased expression of the key anti-oxidants enzymes, decreased inflammatory mediators in islets and conferred protection against oxidative stress in beta cells.
Highlights
Nuclear factor erythroid2-related factor1 (Nrf2) nuclear translocation was confirmed by con-focal microscope within 2 hours after treatment, which was associated with a dose-dependent increase in mRNA expression of antioxidants, including NQO1, HO-1, and GCLC
After induction of oxidative stress with hydrogen peroxide (200 μM) the proportion of dh404-treated viable islet cells was significantly higher in the dh404-treated than untreated islets (74% vs.57%; P
Type 1 diabetes mellitus (T1DM) is an autoimmune disease associated with selected genetic HLA alleles, which result in the permanent destruction of β-cells of the pancreatic islets of Langerhans [1]
Summary
Type 1 diabetes mellitus (T1DM) is an autoimmune disease associated with selected genetic HLA alleles, which result in the permanent destruction of β-cells of the pancreatic islets of Langerhans [1]. Pancreatic islets contain very low levels of the antioxidant enzymes [5]. They have an innate vulnerability to oxidative stress and inflammation [6]. The cytoprotective effects of Nrf are mediated by transcriptional up-regulation of genes encoding numerous antioxidant, detoxifying and cytoprotective enzymes and related molecules. Pancreatic islets are known to contain low level of antioxidants that renders them vulnerable to oxidative stress. Nrf is the master regulator of numerous genes, encoding antioxidant, detoxifying, and cytoprotective molecules. Activation of Nrf pathway induces up-regulation of numerous genes encoding antioxidant and phase II detoxifying enzymes and related proteins. The aim was to investigate the effect of Nrf activator (dh404, CDDO-9,11-dihydro-trifluoroethyl amide) on human islet cells
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