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. 1995 Aug;96(2):1145–1151. doi: 10.1172/JCI118102

Sickle erythrocytes, after sickling, regulate the expression of the endothelin-1 gene and protein in human endothelial cells in culture.

M Phelan 1, S P Perrine 1, M Brauer 1, D V Faller 1
PMCID: PMC185305  PMID: 7635951

Abstract

The molecular defect in sickle cell disease resides in the beta globin gene, with consequent defects in erythrocytes only, suggesting that the vascular occlusion and vasomotor instability which characterize this disease are the result of interactions between abnormal sickle erythrocytes and cells of the blood vessel wall. We explored whether sickle erythrocytes may have effects on vascular tone, exclusive of adhesion events. Exposure of human endothelial cells in culture to previously sickled sickle erythrocytes resulted in a four to eight-fold transcriptional induction of the gene encoding the potent vasoconstrictor endothelin-1 (ET-1). Unsickled sickle erythrocytes or normal erythrocytes exposed to "sickling" conditions had no effect on ET-1 gene induction. Contact of the sickled erythrocytes with the endothelium was not required. Elevations in the ET-1 transcript peaked at 3 h after exposure and persisted for up to 24 h. Four to sixfold increases in the amount of ET-1 peptide was released into the medium surrounding the endothelial cells after exposure to sickled sickle erythrocytes. This is the first demonstration of the regulation of gene expression in endothelial cells as a result of interaction with sickle cells, with induction of genes encoding vasoconstrictors. Furthermore, these findings suggest that sickle erythrocytes may have the capacity to affect local vasomotor tone directly.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Boros L., Thomas C., Weiner W. J. Large cerebral vessel disease in sickle cell anaemia. J Neurol Neurosurg Psychiatry. 1976 Dec;39(12):1236–1239. doi: 10.1136/jnnp.39.12.1236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  3. Cooke J. P., Rossitch E., Jr, Andon N. A., Loscalzo J., Dzau V. J. Flow activates an endothelial potassium channel to release an endogenous nitrovasodilator. J Clin Invest. 1991 Nov;88(5):1663–1671. doi: 10.1172/JCI115481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DIGGS L. W., VORDER BRUEGGE C. F. Vascular occlusive mechanisms in sickle cell disease. J Natl Med Assoc. 1954 Jan;46(1):46–49. [PMC free article] [PubMed] [Google Scholar]
  5. Fabry M. E., Rajanayagam V., Fine E., Holland S., Gore J. C., Nagel R. L., Kaul D. K. Modeling sickle cell vasoocclusion in the rat leg: quantification of trapped sickle cells and correlation with 31P metabolic and 1H magnetic resonance imaging changes. Proc Natl Acad Sci U S A. 1989 May;86(10):3808–3812. doi: 10.1073/pnas.86.10.3808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Frangos J. A., Eskin S. G., McIntire L. V., Ives C. L. Flow effects on prostacyclin production by cultured human endothelial cells. Science. 1985 Mar 22;227(4693):1477–1479. doi: 10.1126/science.3883488. [DOI] [PubMed] [Google Scholar]
  7. Hakim T. S. Flow-induced release of EDRF in the pulmonary vasculature: site of release and action. Am J Physiol. 1994 Jul;267(1 Pt 2):H363–H369. doi: 10.1152/ajpheart.1994.267.1.H363. [DOI] [PubMed] [Google Scholar]
  8. Hatch F. E., Crowe L. R., Miles D. E., Young J. P., Portner M. E. Altered vascular reactivity in sickle hemoglobinopathy. A possible protective factor from hypertension. Am J Hypertens. 1989 Jan;2(1):2–8. doi: 10.1093/ajh/2.1.2. [DOI] [PubMed] [Google Scholar]
  9. Hebbel R. P. Beyond hemoglobin polymerization: the red blood cell membrane and sickle disease pathophysiology. Blood. 1991 Jan 15;77(2):214–237. [PubMed] [Google Scholar]
  10. Hebbel R. P., Boogaerts M. A., Eaton J. W., Steinberg M. H. Erythrocyte adherence to endothelium in sickle-cell anemia. A possible determinant of disease severity. N Engl J Med. 1980 May 1;302(18):992–995. doi: 10.1056/NEJM198005013021803. [DOI] [PubMed] [Google Scholar]
  11. Hebbel R. P., Eaton J. W., Balasingam M., Steinberg M. H. Spontaneous oxygen radical generation by sickle erythrocytes. J Clin Invest. 1982 Dec;70(6):1253–1259. doi: 10.1172/JCI110724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hebbel R. P., Morgan W. T., Eaton J. W., Hedlund B. E. Accelerated autoxidation and heme loss due to instability of sickle hemoglobin. Proc Natl Acad Sci U S A. 1988 Jan;85(1):237–241. doi: 10.1073/pnas.85.1.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hebbel R. P., Yamada O., Moldow C. F., Jacob H. S., White J. G., Eaton J. W. Abnormal adherence of sickle erythrocytes to cultured vascular endothelium: possible mechanism for microvascular occlusion in sickle cell disease. J Clin Invest. 1980 Jan;65(1):154–160. doi: 10.1172/JCI109646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hoover R., Rubin R., Wise G., Warren R. Adhesion of normal and sickle erythrocytes to endothelial monolayer cultures. Blood. 1979 Oct;54(4):872–876. [PubMed] [Google Scholar]
  15. Kaul D. K., Fabry M. E., Nagel R. L. Microvascular sites and characteristics of sickle cell adhesion to vascular endothelium in shear flow conditions: pathophysiological implications. Proc Natl Acad Sci U S A. 1989 May;86(9):3356–3360. doi: 10.1073/pnas.86.9.3356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kennedy A. P., Williams B., Meydrech E. F., Steinberg M. H. Regional and temporal variation in oscillatory blood flow in sickle cell disease. Am J Hematol. 1988 Jun;28(2):92–94. doi: 10.1002/ajh.2830280205. [DOI] [PubMed] [Google Scholar]
  17. Konotey-Ahulu F. I. The sickle cell diseases. Clinical manifestations including the "sickle crisis". Arch Intern Med. 1974 Apr;133(4):611–619. [PubMed] [Google Scholar]
  18. Kourembanas S., Faller D. V. Platelet-derived growth factor production by human umbilical vein endothelial cells is regulated by basic fibroblast growth factor. J Biol Chem. 1989 Mar 15;264(8):4456–4459. [PubMed] [Google Scholar]
  19. Kourembanas S., Hannan R. L., Faller D. V. Oxygen tension regulates the expression of the platelet-derived growth factor-B chain gene in human endothelial cells. J Clin Invest. 1990 Aug;86(2):670–674. doi: 10.1172/JCI114759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kourembanas S., Marsden P. A., McQuillan L. P., Faller D. V. Hypoxia induces endothelin gene expression and secretion in cultured human endothelium. J Clin Invest. 1991 Sep;88(3):1054–1057. doi: 10.1172/JCI115367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kourembanas S., McQuillan L. P., Leung G. K., Faller D. V. Nitric oxide regulates the expression of vasoconstrictors and growth factors by vascular endothelium under both normoxia and hypoxia. J Clin Invest. 1993 Jul;92(1):99–104. doi: 10.1172/JCI116604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kuchan M. J., Frangos J. A. Shear stress regulates endothelin-1 release via protein kinase C and cGMP in cultured endothelial cells. Am J Physiol. 1993 Jan;264(1 Pt 2):H150–H156. doi: 10.1152/ajpheart.1993.264.1.H150. [DOI] [PubMed] [Google Scholar]
  23. Lee M. E., Dhadly M. S., Temizer D. H., Clifford J. A., Yoshizumi M., Quertermous T. Regulation of endothelin-1 gene expression by Fos and Jun. J Biol Chem. 1991 Oct 5;266(28):19034–19039. [PubMed] [Google Scholar]
  24. Lipowsky H. H., Sheikh N. U., Katz D. M. Intravital microscopy of capillary hemodynamics in sickle cell disease. J Clin Invest. 1987 Jul;80(1):117–127. doi: 10.1172/JCI113036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lubin B., Chiu D., Bastacky J., Roelofsen B., Van Deenen L. L. Abnormalities in membrane phospholipid organization in sickled erythrocytes. J Clin Invest. 1981 Jun;67(6):1643–1649. doi: 10.1172/JCI110200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. MacCumber M. W., Ross C. A., Glaser B. M., Snyder S. H. Endothelin: visualization of mRNAs by in situ hybridization provides evidence for local action. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7285–7289. doi: 10.1073/pnas.86.18.7285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Malek A. M., Gibbons G. H., Dzau V. J., Izumo S. Fluid shear stress differentially modulates expression of genes encoding basic fibroblast growth factor and platelet-derived growth factor B chain in vascular endothelium. J Clin Invest. 1993 Oct;92(4):2013–2021. doi: 10.1172/JCI116796. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Malek A. M., Greene A. L., Izumo S. Regulation of endothelin 1 gene by fluid shear stress is transcriptionally mediated and independent of protein kinase C and cAMP. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):5999–6003. doi: 10.1073/pnas.90.13.5999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Merkel K. H., Ginsberg P. L., Parker J. C., Jr, Post M. J. Cerebrovascular disease in sickle cell anemia: a clinical, pathological and radiological correlation. Stroke. 1978 Jan-Feb;9(1):45–52. doi: 10.1161/01.str.9.1.45. [DOI] [PubMed] [Google Scholar]
  30. Meyer M., Pahl H. L., Baeuerle P. A. Regulation of the transcription factors NF-kappa B and AP-1 by redox changes. Chem Biol Interact. 1994 Jun;91(2-3):91–100. doi: 10.1016/0009-2797(94)90029-9. [DOI] [PubMed] [Google Scholar]
  31. Middelkoop E., Lubin B. H., Bevers E. M., Op den Kamp J. A., Comfurius P., Chiu D. T., Zwaal R. F., van Deenen L. L., Roelofsen B. Studies on sickled erythrocytes provide evidence that the asymmetric distribution of phosphatidylserine in the red cell membrane is maintained by both ATP-dependent translocation and interaction with membrane skeletal proteins. Biochim Biophys Acta. 1988 Jan 22;937(2):281–288. doi: 10.1016/0005-2736(88)90250-7. [DOI] [PubMed] [Google Scholar]
  32. Miller V. M., Burnett J. C., Jr Modulation of NO and endothelin by chronic increases in blood flow in canine femoral arteries. Am J Physiol. 1992 Jul;263(1 Pt 2):H103–H108. doi: 10.1152/ajpheart.1992.263.1.H103. [DOI] [PubMed] [Google Scholar]
  33. Milner P., Bodin P., Loesch A., Burnstock G. Increased shear stress leads to differential release of endothelin and ATP from isolated endothelial cells from 4- and 12-month-old male rabbit aorta. J Vasc Res. 1992 Nov-Dec;29(6):420–425. doi: 10.1159/000158960. [DOI] [PubMed] [Google Scholar]
  34. Milner P., Bodin P., Loesch A., Burnstock G. Rapid release of endothelin and ATP from isolated aortic endothelial cells exposed to increased flow. Biochem Biophys Res Commun. 1990 Jul 31;170(2):649–656. doi: 10.1016/0006-291x(90)92141-l. [DOI] [PubMed] [Google Scholar]
  35. Mitsumata M., Fishel R. S., Nerem R. M., Alexander R. W., Berk B. C. Fluid shear stress stimulates platelet-derived growth factor expression in endothelial cells. Am J Physiol. 1993 Jul;265(1 Pt 2):H3–H8. doi: 10.1152/ajpheart.1993.265.1.H3. [DOI] [PubMed] [Google Scholar]
  36. Moon D. G., Horgan M. J., Andersen T. T., Krystek S. R., Jr, Fenton J. W., 2nd, Malik A. B. Endothelin-like pulmonary vasoconstrictor peptide release by alpha-thrombin. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9529–9533. doi: 10.1073/pnas.86.23.9529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Morita T., Kurihara H., Maemura K., Yoshizumi M., Yazaki Y. Disruption of cytoskeletal structures mediates shear stress-induced endothelin-1 gene expression in cultured porcine aortic endothelial cells. J Clin Invest. 1993 Oct;92(4):1706–1712. doi: 10.1172/JCI116757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Mosseri M., Bartlett-Pandite A. N., Wenc K., Isner J. M., Weinstein R. Inhibition of endothelium-dependent vasorelaxation by sickle erythrocytes. Am Heart J. 1993 Aug;126(2):338–346. doi: 10.1016/0002-8703(93)91049-k. [DOI] [PubMed] [Google Scholar]
  39. Nishida K., Harrison D. G., Navas J. P., Fisher A. A., Dockery S. P., Uematsu M., Nerem R. M., Alexander R. W., Murphy T. J. Molecular cloning and characterization of the constitutive bovine aortic endothelial cell nitric oxide synthase. J Clin Invest. 1992 Nov;90(5):2092–2096. doi: 10.1172/JCI116092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Pohl U., Herlan K., Huang A., Bassenge E. EDRF-mediated shear-induced dilation opposes myogenic vasoconstriction in small rabbit arteries. Am J Physiol. 1991 Dec;261(6 Pt 2):H2016–H2023. doi: 10.1152/ajpheart.1991.261.6.H2016. [DOI] [PubMed] [Google Scholar]
  41. Rodgers G. P., Schechter A. N., Noguchi C. T., Klein H. G., Nienhuis A. W., Bonner R. F. Periodic microcirculatory flow in patients with sickle-cell disease. N Engl J Med. 1984 Dec 13;311(24):1534–1538. doi: 10.1056/NEJM198412133112403. [DOI] [PubMed] [Google Scholar]
  42. Schacter L. P. Generation of superoxide anion and hydrogen peroxide by erythrocytes from individuals with sickle trait or normal haemoglobin. Eur J Clin Invest. 1986 Jun;16(3):204–210. doi: 10.1111/j.1365-2362.1986.tb01330.x. [DOI] [PubMed] [Google Scholar]
  43. Schenk H., Klein M., Erdbrügger W., Dröge W., Schulze-Osthoff K. Distinct effects of thioredoxin and antioxidants on the activation of transcription factors NF-kappa B and AP-1. Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1672–1676. doi: 10.1073/pnas.91.5.1672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Stockman J. A., Nigro M. A., Mishkin M. M., Oski F. A. Occlusion of large cerebral vessels in sickle-cell anemia. N Engl J Med. 1972 Oct 26;287(17):846–849. doi: 10.1056/NEJM197210262871703. [DOI] [PubMed] [Google Scholar]
  45. Tajika T., Ono Y., Ono K., Miura M., Kyo Y. The effect of probucol on the proliferation of cultured human umbilical vascular endothelial cells. In Vitro Cell Dev Biol Anim. 1993 May;29A(5):347–349. doi: 10.1007/BF02633978. [DOI] [PubMed] [Google Scholar]
  46. Tesfamariam B., Cohen R. A. Free radicals mediate endothelial cell dysfunction caused by elevated glucose. Am J Physiol. 1992 Aug;263(2 Pt 2):H321–H326. doi: 10.1152/ajpheart.1992.263.2.H321. [DOI] [PubMed] [Google Scholar]
  47. Vane J. R., Anggård E. E., Botting R. M. Regulatory functions of the vascular endothelium. N Engl J Med. 1990 Jul 5;323(1):27–36. doi: 10.1056/NEJM199007053230106. [DOI] [PubMed] [Google Scholar]
  48. Wautier J. L., Pintigny D., Maclouf J., Wautier M. P., Corvazier E., Caen J. Release of prostacyclin after erythrocyte adhesion to cultured vascular endothelium. J Lab Clin Med. 1986 Mar;107(3):210–215. [PubMed] [Google Scholar]
  49. Weinstein R., Zhou M. A., Bartlett-Pandite A., Wenc K. Sickle erythrocytes inhibit human endothelial cell DNA synthesis. Blood. 1990 Nov 15;76(10):2146–2152. [PubMed] [Google Scholar]
  50. Yanagisawa M., Kurihara H., Kimura S., Tomobe Y., Kobayashi M., Mitsui Y., Yazaki Y., Goto K., Masaki T. A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature. 1988 Mar 31;332(6163):411–415. doi: 10.1038/332411a0. [DOI] [PubMed] [Google Scholar]

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