Hypoxia-reoxygenation inhibits gap junctional communication in cultured human umbilical vein endothelial cells.
We studied the change in gap junctional intercellular communication (GJIC) on human umbilical vein endothelial cells (HUVEC) under hypoxia-reoxygenation (H-R) conditions by the fluorescence redistribution after photobleaching (FRAP) method. Confluent HUVEC monolayers were exposed to hypoxia (pO2<0.1%) for 12 hours, and then were returned to normal atmospheric conditions for reoxygenation. Contrast microscopic observation showed no significant changes in the morphology of the HUVEC at any times after H-R. Reoxygenation following hypoxia caused time-dependent decrease in GJIC, that is, GJIC reduction was induced after 2 hours and reached maximum at 4-6 hours which recovered to normal levels after 18 hours. Oxidant sensitive fluorescence dye assay revealed that the generation of intracellular free radicals increased during the first 2 hours after reoxygenation. Hydroxyl radical scavengers (MCI-186, DMSO) and an iron chelator (deferoxamine) abolished the reduction of GJIC due to H-R. However, SOD, catalase and probucol were essentially inactive on this reduction. These data suggest that ischemia-reperfusion injury may be caused by a functional defect of GJIC induced by reactive oxygen radicals.
Nishida
M
M
Department of Cellular Physiological Chemistry, Graduate School, Tokyo Medical and Dental University, Japan.
Futami
S
S
Morita
I
I
Maekawa
K
K
Murota
S I
SI
D000983:Antipyrine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002374:Catalase / Q000378:metabolism; D002450:Cell Communication; D015687:Cell Hypoxia; D002478:Cells, Cultured; D003676:Deferoxamine / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D000077553:Edaravone; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D017629:Gap Junctions / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D019044:Image Cytometry / Q000379:methods; D007834:Lasers; D010100:Oxygen / Q000502:physiology*; D011341:Probucol / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D013997:Time Factors; D014471:Umbilical Veins / Q000166:cytology / Q000502:physiology
D017382:Reactive Oxygen Species; D002374:Catalase; D013482:Superoxide Dismutase; D003676:Deferoxamine; D011341:Probucol; D000077553:Edaravone; D010100:Oxygen; D000983:Antipyrine; D004121:Dimethyl Sulfoxide