Endothelial gap junctions are down-regulated by arsenic trioxide

Abstract

We investigated the effect of As 2O 3, an anti-cancer drug, on endothelial gap junctions. Human aortic endothelial cells (HAEC) were treated with As 2O 3 at 1, 10, 100, and 1000 ng/ml and the cells were examined to evaluate the expression of connexin43 (Cx43) and to assess gap-junction communication. Endothelial nitric oxide synthase (eNOS) and nitric oxide (NO) were measured to assess for endothelial dysfunction. Male Sprague–Dawley rats were given intravenous As 2O 3 (200 μg/kg/day) or saline for 4 weeks, after which aortic endothelial gap junctions, eNOS, and circulating NO levels were evaluated. We found that HAEC Cx43 transcripts and gap junctions were reduced and gap-junction communication was attenuated by As 2O 3. This decrease of Cx43 gap junctions was prevented by the addition of protease inhibitors. At a dose of 100 ng/ml of As 2O 3, eNOS was reduced at 48 h, but NO was markedly reduced by 1 h. In animals treated with As 2O 3, endothelial gap junctions comprising Cx37, Cx40, or Cx43 were all reduced in amount, while eNOS and circulating NO levels remained unchanged. In both in vitro and in vivo rat experiments, endothelial gap junctions were consistently reduced by As 2O 3, unlike the response of eNOS and NO, which were decreased in cells but not in the rat aortic endothelium. The reduction in Cx43 involved both down-regulation at the transcriptional level and increased degradation. These findings indicate that gap-junction communication in the vascular endothelium is inhibited by treatment with As 2O 3.