Influence of sustained mechanical stress on Egr-1 mRNA expression in cultured human endothelial cells
| Autor: | M, Stula, H D, Orzechowski, S, Gschwend, R, Vetter, R, von Harsdorf, R, Dietz, M, Paul |
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| Rok vydání: | 2000 |
| Předmět: |
Sulfonamides
Umbilical Veins Gadolinium Blotting Northern Isoquinolines Actins Cell Line Immediate-Early Proteins DNA-Binding Proteins Microscopy Fluorescence 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine Humans Endothelium Vascular RNA Messenger Stress Mechanical Enzyme Inhibitors Colchicine Egtazic Acid Protein Kinase Inhibitors Proto-Oncogene Proteins c-fos Cells Cultured Chelating Agents Early Growth Response Protein 1 Signal Transduction Transcription Factors |
| Zdroj: | Molecular and cellular biochemistry. 210(1-2) |
| ISSN: | 0300-8177 |
| Popis: | Restenosis after initially successful balloon angioplasty of coronary artery stenosis remains a major problem in clinical cardiology. Previous studies have identified pathogenetic factors which trigger cell proliferation and vascular remodeling ultimately leading to restenosis. Since there is evidence that endothelial cells adjacent to the angioplasty wound area synthesize factors which may initiate this process, we investigated the effects of mechanical stimulation on endothelial gene expression in vitro and focussed on the influence of sustained mechanical stress on expression of immediate early genes which have previously been shown to be induced in the vascular wall in vivo. Primary cultured human umbilical vein endothelial cells (HUVEC) and the human endothelial cell line EA.hy 926 were plated on collagen-coated silicone membranes and subjected to constant longitudinal stress of approximately 20% for 10 min to 6 h. Total RNA was isolated and the expression of the immediate early genes c-Fos and Egr-1 was studied by Northern blot analysis. We found a rapid upregulation c-Fos and Egr-1 mRNA which started at 10 min and reached its maxima at 30 min. HUVEC lost most of their stretch response after the third passage whereas immediate early gene expression was constantly in EA.hy 926 cells. Using specific inhibitors we investigated the contribution of several signal transduction pathways to stretch-activated Egr-1 mRNA expression. We found significant suppression of stretch-induced Egr-1 mRNA expression by protein kinase C (PKC) inhibition (p0.05) and by calcium depletion (EA.hy 926, p0.05; HUVEC, p = 0.063). No effect on stretch-activated Egr-1 mRNA expression was detected by inhibition of protein kinase A, blockade of stretch-activated cation channels or inhibition of microtubule synthesis. We conclude that sustained mechanical strain induces Egr-1 mRNA expression by PKC- and calcium-dependent mechanisms. |
| Databáze: | OpenAIRE |
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