Effects of thrombin on the integrity of monolayers of cultured human endothelial cells.
51Cr-prelabelled endothelial cells (EC) in confluent monolayers were incubated in RPMI 1640 + foetal calf serum 20% (v/v) to which purified thrombin was added. Thrombin (greater than or equal to 0.1 NIH U/ml) significantly accelerated 51Cr-release and caused extensive but reversible cell "contraction". Thrombin-exposed EC reacted to a new dose of thrombin with no appreciable shape change, but 51Cr-efflux was again accelerated. EC exposed to thrombin pretreated with N-bromosuccinimide (modifying the macromolecular site) or phenylmethylsulfonyl fluoride (blocking the serine site) retained normal morphology and did not leak excess amounts of 51Cr. Antithrombin III also inhibited the effect of thrombin. Pretreatment of EC with either indomethacin, aspirin, sulfinpyrazone, pronase or neuraminidase did not influence the effect of subsequent thrombin exposure.
D000818:Animals; D000990:Antithrombin III / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D002860:Chromium Radioisotopes / Q000378:metabolism; D003470:Culture Media; D004727:Endothelium / Q000166:cytology / Q000187:drug effects; D006629:Hirudins / Q000494:pharmacology; D006801:Humans; D009439:Neuraminidase / Q000494:pharmacology; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D011402:Pronase / Q000494:pharmacology; D014176:Protein Biosynthesis; D013917:Thrombin / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D014158:Transcription, Genetic; D014471:Umbilical Veins / Q000166:cytology* / Q000187:drug effects
D002860:Chromium Radioisotopes; D003470:Culture Media; D006629:Hirudins; D010664:Phenylmethylsulfonyl Fluoride; D000990:Antithrombin III; D009439:Neuraminidase; D013917:Thrombin; D011402:Pronase