high glucose-mediated oxidative stress impairs cell migration高glucose-mediated氧化应激损害细胞迁移.pdfVIP

High Glucose-Mediated Oxidative Stress Impairs Cell Migration 1,2 ´ 2 3 3 Marcelo L. Lamers , Maıra E. S. Almeida , Miguel Vicente-Manzanares , Alan F. Horwitz , Marinilce F. Santos2* 1 Department of Morphological Sciences, Institute of Basic Health Science, Federal University of Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil, 2 Department ˜ ˜ ˜ of Cell and Developmental Biology, Biomedical Sciences Institute, University of Sao Paulo, Sao Paulo, Sao Paulo, Brazil, 3 Department of Cell Biology, University of Virginia, Charlottesville, Virginia, United States of America Abstract Deficient wound healing in diabetic patients is very frequent, but the cellular and molecular causes are poorly defined. In this study, we evaluate the hypothesis that high glucose concentrations inhibit cell migration. Using CHO.K1 cells, NIH-3T3 fibroblasts, mouse embryonic fibroblasts and primary skin fibroblasts from control and diabetic rats cultured in 5 mM D- glucose (low glucose, LG), 25 mM D-glucose (high glucose, HG) or 25 mM L-glucose medium (osmotic control - OC), we analyzed the migration speed, protrusion stability, cell polarity, adhesion maturation and the activity of the small Rho GTPase Rac1. We also analyzed the effects of reactive oxygen species by incubating cells with the antioxidant N-Acetyl- Cysteine (NAC). We observed that HG conditions inhibited cell migration when compared to LG or OC. This inhibition resulted from impaired cell polarity, protrus