title: Fluid flow decreases preproendothelin mRNA levels and suppresses endothelin-1 peptide release in cultured human endothelial cells.
abstract: Endothelin-1, a 21-amino acid peptide secreted by endothelial cells, has constrictor and mitogenic activity for vascular smooth muscle cells, and its mitogenic activity is synergistic with that of platelet-derived growth factor. Endothelial cell-derived endothelin-1 might therefore contribute to intimal hyperplasia in reendothelialized segments of vascular grafts or of endarterectomy and angioplasty sites. Because intimal hyperplasia occurs most often at sites with disordered flow patterns and lower fluid shear stress, we tested the effects of static culture versus high laminar shear stress (25 dyne/cm2) on endothelin-1 precursor (preproendothelin) gene mRNA transcript levels and endothelin-1 peptide release in cultured human endothelial cells. Primary cultures of human umbilical vein endothelial cells were subjected to controlled levels of shear stress in parallel plate flow chambers for 24 hours. To detect preproendothelin mRNA we applied a linked reverse transcriptase-polymerase chain reaction (RT/PCR) to RNA extracted from cultures. Southern blots of RT/PCR reaction products were hybridized with radioactive phosphorous (32P) labeled probes for the amplified preproendothelin complementary deoxyribonucleic acid (cDNA). Detection by RT/PCR of mRNA for glyceraldehyde 3-phosphate dehydrogenase was used to measure a constitutively expressed control signal. Endothelin-1 release into culture medium was measured by radioimmunoassay. Application of 25 dyne/cm2 of shear stress for 24 hours sharply reduced endothelial cell levels of precursor preproendothelin mRNA.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Surgery, Uniformed Services University of the Health Sciences, Bethesda, Md.
authors.forename: J B
authors.initials: JB
authors.lastname: Sharefkin
authors.forename: S L
authors.initials: SL
authors.lastname: Diamond
authors.forename: S G
authors.initials: SG
authors.lastname: Eskin
authors.forename: L V
authors.initials: LV
authors.lastname: McIntire
authors.forename: C W
authors.initials: CW
authors.lastname: Dieffenbach
mesh_terms: D015139:Blotting, Southern; D002478:Cells, Cultured; D019332:Endothelin-1; D016232:Endothelins / Q000235:genetics* / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D011498:Protein Precursors / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012212:Rheology; D014158:Transcription, Genetic
chemical_list: D019332:Endothelin-1; D016232:Endothelins; D015345:Oligonucleotide Probes; D011498:Protein Precursors; D012333:RNA, Messenger