Endothelial gap junctions are down-regulated by arsenic trioxide.
We investigated the effect of As(2)O(3), an anti-cancer drug, on endothelial gap junctions. Human aortic endothelial cells (HAEC) were treated with As(2)O(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(2)O(3) (200 mug/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(2)O(3). This decrease of Cx43 gap junctions was prevented by the addition of protease inhibitors. At a dose of 100 ng/ml of As(2)O(3), eNOS was reduced at 48 h, but NO was markedly reduced by 1 h. In animals treated with As(2)O(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(2)O(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(2)O(3).
Chou
Yusan
Y
Department of Internal Medicine, Mackay Memorial Hospital, Taipei, Taiwan.
Tsai
Cheng-Ho
CH
Ueng
Kwo-Chang
KC
Tian
Tin-Yi
TY
Chen
Shu-Chen
SC
Yeh
Hung-I
HI
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D000077237:Arsenic Trioxide; D001152:Arsenicals / Q000494:pharmacology*; D015153:Blotting, Western; D002460:Cell Line; D018031:Connexin 43 / Q000235:genetics / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D017629:Gap Junctions / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007976:Leupeptins / Q000494:pharmacology; D008297:Male; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism; D010087:Oxides / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014841:von Willebrand Factor / Q000235:genetics / Q000378:metabolism
D000970:Antineoplastic Agents; D001152:Arsenicals; D018031:Connexin 43; D015853:Cysteine Proteinase Inhibitors; D007976:Leupeptins; D010087:Oxides; D012333:RNA, Messenger; D014841:von Willebrand Factor; C063261:acetylleucyl-leucyl-norleucinal; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; C032854:leupeptin; D000077237:Arsenic Trioxide