RESUMEN
In cultured human umbilical vein endothelial cells (HUVECs) high glucose (HG) stimulation will lead to significant cell death. Bardoxolone-methyl (BARD) is a NF-E2 p45-related factor 2 (Nrf2) agonist. In this study we show that BARD, at only nM concentrations, activated Nrf2 signaling in HUVECs. BARD induced Keap1-Nrf2 disassociation, Nrf2 protein stabilization and nuclear translocation, increasing expression of antioxidant response element (ARE) genes. BARD pretreatment in HUVECs inhibited HG-induced reactive oxygen species production, oxidative injury and cell apoptosis. Nrf2 shRNA or knockout (using a CRISPR/Cas9 construct) reversed BARD-induced cytoprotection in HG-stimulated HUVECs. Conversely, forced activation of Nrf2 cascade by Keap1 shRNA mimicked BARD's activity and protected HUVECs from HG. Importantly, BARD failed to offer further cytoprotection against HG in the Keap1-silened HUVECs. Taken together, Keap1-Nrf2 cascade activation by BARD protects HUVECs from HG-induced oxidative injury.
Asunto(s)
Angiopatías Diabéticas/prevención & control , Hiperglucemia/complicaciones , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Factor 2 Relacionado con NF-E2/agonistas , Ácido Oleanólico/análogos & derivados , Estrés Oxidativo/efectos de los fármacos , Apoptosis/efectos de los fármacos , Glucemia/metabolismo , Angiopatías Diabéticas/etiología , Angiopatías Diabéticas/metabolismo , Evaluación Preclínica de Medicamentos , Células Endoteliales de la Vena Umbilical Humana , Humanos , Factor 2 Relacionado con NF-E2/metabolismo , Ácido Oleanólico/farmacología , Ácido Oleanólico/uso terapéutico , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
microRNAs are a class of short non-coding RNAs containing about 19-22 nucleotides that regulate target gene expression through post-transcriptional repression or mRNA degradation, and involved in a variety of biological processes, such as cellular differentiation, proliferation, apoptosis and metabolism. microRNA-182 (miR-182), belonging to miR-183/96/182 cluster that consists of miR-182, -183, and -96, highly expresses in many cells and tissues, including osteoblasts, lymphocytes, adipocytes, retina, inner ear, etc. The recent studies of miR-182 highlighted its multiple important roles in differentiation, development, and functional maintenance in the cells and tissues. The dysregulation of miR-182 is associated with occurrence and development of many diseases, such as retinopathy, autoimmune diseases, cancers, obesity and diabetes. This review summarizes recent research progresses on the roles and mechanisms of miR-182 in cellular function and diseases.