Oxidative Stress-Responsive Transcription Factor ATF3 Potentially Mediates Diabetic Angiopathy

Abstract

Previous results of our cDNA microarray analysis to look for genes whose expression level correlates well with in vitro tubulogenesis by NP31 endothelial cells revealed the transcription factor ATF3 known to be responsive to stress such as reactive oxygen species (ROS). Anti-ATF3 small interfering RNA gave an inhibitory influence on tube formation by NP31 cells expressing an activated form of the vascular endothelial growth factor receptor 1 (VEGFR-1) kinase. When expression of ATF3 was regulated under the control of tetracycline system in NP31 cells, they acquired the tubulogenic ability upon ATF3 induction. While ATF3 failed to induce expressions of VEGF and VEGFR, it regulated those of CDK2, CDK4, p8, plasminogen activator inhibitor 1, integrin α1, subunit and matrix metalloprotease MMP13. In H₂O₂-stimulated NP31 cells as well as endothelial cells of glomerulus and aorta of Otsuka-Long-Evans-Tokushima-Fatty diabetic model rats, concomitantly enhanced expressions of ATF3, PAI-1, and p8 were observed. Given the proposed hypothesis of the close linkage between diabetic angiopathy and ROS, those data suggest that ROS-associated diabetic complication may involve ATF3-mediated pathological angiogenesis.

We thank O. N. Witte at the University of California—Los Angeles for the Tet constructs, Otsuka Pharmaceutical Co. for the OLETF animals, and S. Kobayashi for technical help. We also thank S. Fujii at Hokkaido University for providing PAI-1 promoter constructs and S. Morikawa for technical help in confocal microscopy.

This work is supported by a grant-in-aid for scientific research on metal sensors (no. 12147210) from the Japanese government.