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Logo of jcinvestThe Journal of Clinical Investigation
J Clin Invest. 1995 August; 96(2): 1145–1151.
PMCID: PMC185305

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


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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  • DIGGS LW, VORDER BRUEGGE CF. Vascular occlusive mechanisms in sickle cell disease. J Natl Med Assoc. 1954 Jan;46(1):46–49. [PMC free article] [PubMed]
  • Konotey-Ahulu FI. The sickle cell diseases. Clinical manifestations including the "sickle crisis". Arch Intern Med. 1974 Apr;133(4):611–619. [PubMed]
  • Boros L, Thomas C, Weiner WJ. Large cerebral vessel disease in sickle cell anaemia. J Neurol Neurosurg Psychiatry. 1976 Dec;39(12):1236–1239. [PMC free article] [PubMed]
  • Merkel KH, Ginsberg PL, Parker JC, Jr, Post MJ. Cerebrovascular disease in sickle cell anemia: a clinical, pathological and radiological correlation. Stroke. 1978 Jan-Feb;9(1):45–52. [PubMed]
  • Stockman JA, Nigro MA, Mishkin MM, Oski FA. Occlusion of large cerebral vessels in sickle-cell anemia. N Engl J Med. 1972 Oct 26;287(17):846–849. [PubMed]
  • 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]
  • Hebbel RP, Yamada O, Moldow CF, Jacob HS, White JG, Eaton JW. 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. [PMC free article] [PubMed]
  • Hebbel RP, Boogaerts MA, Eaton JW, Steinberg MH. 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. [PubMed]
  • Wautier JL, Pintigny D, Maclouf J, Wautier MP, 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]
  • Weinstein R, Zhou MA, Bartlett-Pandite A, Wenc K. Sickle erythrocytes inhibit human endothelial cell DNA synthesis. Blood. 1990 Nov 15;76(10):2146–2152. [PubMed]
  • Vane JR, Anggård EE, Botting RM. Regulatory functions of the vascular endothelium. N Engl J Med. 1990 Jul 5;323(1):27–36. [PubMed]
  • 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. [PubMed]
  • MacCumber MW, Ross CA, Glaser BM, Snyder SH. 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. [PubMed]
  • Moon DG, Horgan MJ, Andersen TT, Krystek SR, Jr, Fenton JW, 2nd, Malik AB. Endothelin-like pulmonary vasoconstrictor peptide release by alpha-thrombin. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9529–9533. [PubMed]
  • 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. [PubMed]
  • Kourembanas S, McQuillan LP, Leung GK, Faller DV. 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. [PMC free article] [PubMed]
  • Kourembanas S, Marsden PA, McQuillan LP, Faller DV. Hypoxia induces endothelin gene expression and secretion in cultured human endothelium. J Clin Invest. 1991 Sep;88(3):1054–1057. [PMC free article] [PubMed]
  • Kourembanas S, Faller DV. 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]
  • Kourembanas S, Hannan RL, Faller DV. 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. [PMC free article] [PubMed]
  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed]
  • Hebbel RP, Eaton JW, Balasingam M, Steinberg MH. Spontaneous oxygen radical generation by sickle erythrocytes. J Clin Invest. 1982 Dec;70(6):1253–1259. [PMC free article] [PubMed]
  • Schacter LP. 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. [PubMed]
  • Hebbel RP, Morgan WT, Eaton JW, Hedlund BE. Accelerated autoxidation and heme loss due to instability of sickle hemoglobin. Proc Natl Acad Sci U S A. 1988 Jan;85(1):237–241. [PubMed]
  • 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. [PubMed]
  • Tesfamariam B, Cohen RA. Free radicals mediate endothelial cell dysfunction caused by elevated glucose. Am J Physiol. 1992 Aug;263(2 Pt 2):H321–H326. [PubMed]
  • Lee ME, Dhadly MS, Temizer DH, Clifford JA, 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]
  • Meyer M, Pahl HL, Baeuerle PA. Regulation of the transcription factors NF-kappa B and AP-1 by redox changes. Chem Biol Interact. 1994 Jun;91(2-3):91–100. [PubMed]
  • 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. [PubMed]
  • Kennedy AP, Williams B, Meydrech EF, Steinberg MH. Regional and temporal variation in oscillatory blood flow in sickle cell disease. Am J Hematol. 1988 Jun;28(2):92–94. [PubMed]
  • Rodgers GP, Schechter AN, Noguchi CT, Klein HG, Nienhuis AW, Bonner RF. Periodic microcirculatory flow in patients with sickle-cell disease. N Engl J Med. 1984 Dec 13;311(24):1534–1538. [PubMed]
  • Lipowsky HH, Sheikh NU, Katz DM. Intravital microscopy of capillary hemodynamics in sickle cell disease. J Clin Invest. 1987 Jul;80(1):117–127. [PMC free article] [PubMed]
  • Hatch FE, Crowe LR, Miles DE, Young JP, Portner ME. Altered vascular reactivity in sickle hemoglobinopathy. A possible protective factor from hypertension. Am J Hypertens. 1989 Jan;2(1):2–8. [PubMed]
  • Mosseri M, Bartlett-Pandite AN, Wenc K, Isner JM, Weinstein R. Inhibition of endothelium-dependent vasorelaxation by sickle erythrocytes. Am Heart J. 1993 Aug;126(2):338–346. [PubMed]
  • Hebbel RP. Beyond hemoglobin polymerization: the red blood cell membrane and sickle disease pathophysiology. Blood. 1991 Jan 15;77(2):214–237. [PubMed]
  • 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. [PubMed]
  • Kuchan MJ, Frangos JA. 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. [PubMed]
  • Malek AM, Greene AL, 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. [PubMed]
  • Mitsumata M, Fishel RS, Nerem RM, Alexander RW, Berk BC. Fluid shear stress stimulates platelet-derived growth factor expression in endothelial cells. Am J Physiol. 1993 Jul;265(1 Pt 2):H3–H8. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Malek AM, Gibbons GH, Dzau VJ, 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. [PMC free article] [PubMed]
  • Hakim TS. 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. [PubMed]
  • Miller VM, Burnett JC., 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. [PubMed]
  • Cooke JP, Rossitch E, Jr, Andon NA, Loscalzo J, Dzau VJ. Flow activates an endothelial potassium channel to release an endogenous nitrovasodilator. J Clin Invest. 1991 Nov;88(5):1663–1671. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Nishida K, Harrison DG, Navas JP, Fisher AA, Dockery SP, Uematsu M, Nerem RM, Alexander RW, Murphy TJ. Molecular cloning and characterization of the constitutive bovine aortic endothelial cell nitric oxide synthase. J Clin Invest. 1992 Nov;90(5):2092–2096. [PMC free article] [PubMed]
  • Frangos JA, Eskin SG, McIntire LV, Ives CL. Flow effects on prostacyclin production by cultured human endothelial cells. Science. 1985 Mar 22;227(4693):1477–1479. [PubMed]
  • Kaul DK, Fabry ME, Nagel RL. 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. [PubMed]
  • Fabry ME, Rajanayagam V, Fine E, Holland S, Gore JC, Nagel RL, Kaul DK. 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. [PubMed]
  • Lubin B, Chiu D, Bastacky J, Roelofsen B, Van Deenen LL. Abnormalities in membrane phospholipid organization in sickled erythrocytes. J Clin Invest. 1981 Jun;67(6):1643–1649. [PMC free article] [PubMed]
  • Middelkoop E, Lubin BH, Bevers EM, Op den Kamp JA, Comfurius P, Chiu DT, Zwaal RF, van Deenen LL, 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. [PubMed]

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