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. 1995 May;146(5):1065–1072.

Up-regulation of endothelin-1 mRNA and peptide expression in rat cardiac allografts with rejection and arteriosclerosis.

B Watschinger 1, M H Sayegh 1, W W Hancock 1, M E Russell 1
PMCID: PMC1869298  PMID: 7747801

Abstract

Acute and chronic rejection are frequent and significant complications of cardiac transplantation, and graft arteriosclerosis is the leading cause of death beyond the first year after transplant. Levels of endothelin-1 (ET-1) are elevated in plasma of patients with cardiac allografts and those with symptomatic vascular atherosclerosis, but little is known about the role of ET-1 in these processes. This study examined intragraft ET-1 expression in rat cardiac models of acute rejection and chronic rejection associated with graft arteriosclerosis. Corrected ET-1 gene transcript levels were measured with a [32P]dCTP reverse transcription polymerase chain reaction assay normalized with glyceraldehyde-3-phosphate dehydrogenase, and the gene product was evaluated by immunohistology with a monospecific anti-ET-1 antibody at different time points after transplant. ET-1 mRNA levels were significantly increased in acutely rejected (Wistar-Furth rat cardiac allografts transplanted into Lewis rat recipients) and chronically rejected (Lewis allografts transplanted into F344 recipients) vascularized cardiac allografts as compared with isograft controls. In acutely rejected allografts, peak expression occurred on day 5 after transplant. In chronically rejected allografts, the increase in ET-1 mRNA was sustained on days 7, 28, and 75. In both acutely and chronically rejected allografts, ET-1 mRNA upregulation was not seen in host spleens or paired host hearts. Immunohistological analysis confirmed that the bulk of ET-1 peptide expression was localized to mononuclear cells that diffusely infiltrated the graft interstitium (acute rejection and early chronic rejection) and accumulated within the neointima of chronically rejecting hearts with arteriosclerosis. These observations, taken together with in vitro data showing that ET-1 production is stimulated by certain cytokines, indicate that the allogeneic stimulus within rejecting vascularized cardiac allografts, presumably cytokine mediated, leads to significant intragraft up-regulation of ET-1 mRNA and peptide expression. The local up-regulation of this vasoactive and mitogenic peptide within acutely and chronically rejected cardiac allografts suggests that ET-1 may be involved in the development of graft arteriosclerosis.

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Selected References

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  1. Adams D. H., Tilney N. L., Collins J. J., Jr, Karnovsky M. J. Experimental graft arteriosclerosis. I. The Lewis-to-F-344 allograft model. Transplantation. 1992 May;53(5):1115–1119. [PubMed] [Google Scholar]
  2. Adams D. H., Wyner L. R., Karnovsky M. J. Experimental graft arteriosclerosis. II. Immunocytochemical analysis of lesion development. Transplantation. 1993 Oct;56(4):794–799. doi: 10.1097/00007890-199310000-00004. [DOI] [PubMed] [Google Scholar]
  3. Berkenboom G., Unger P., Dequenne P., Marchant A., Goldman M., Antoine M., LeClerc J. L. Effects of serotonin on coronary arteries of cardiac transplant recipients. Am J Cardiol. 1993 Aug 1;72(3):331–335. doi: 10.1016/0002-9149(93)90681-2. [DOI] [PubMed] [Google Scholar]
  4. Bobik A., Grooms A., Millar J. A., Mitchell A., Grinpukel S. Growth factor activity of endothelin on vascular smooth muscle. Am J Physiol. 1990 Mar;258(3 Pt 1):C408–C415. doi: 10.1152/ajpcell.1990.258.3.C408. [DOI] [PubMed] [Google Scholar]
  5. Coughlan A. F., Hau H., Dunlop L. C., Berndt M. C., Hancock W. W. P-selectin and platelet-activating factor mediate initial endotoxin-induced neutropenia. J Exp Med. 1994 Jan 1;179(1):329–334. doi: 10.1084/jem.179.1.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dorent R., Cacoub P., Carayon A., Nataf P., Golmard J. L., Vaissier E., Godeau P., Cabrol C., Gandjbakhch I. Endothelin levels after orthotopic heart transplantation. Transplant Proc. 1994 Feb;26(1):250–250. [PubMed] [Google Scholar]
  7. Douglas S. A., Louden C., Vickery-Clark L. M., Storer B. L., Hart T., Feuerstein G. Z., Elliott J. D., Ohlstein E. H. A role for endogenous endothelin-1 in neointimal formation after rat carotid artery balloon angioplasty. Protective effects of the novel nonpeptide endothelin receptor antagonist SB 209670. Circ Res. 1994 Jul;75(1):190–197. doi: 10.1161/01.res.75.1.190. [DOI] [PubMed] [Google Scholar]
  8. Ehrenreich H., Burd P. R., Rottem M., Hültner L., Hylton J. B., Garfield M., Coligan J. E., Metcalfe D. D., Fauci A. S. Endothelins belong to the assortment of mast cell-derived and mast cell-bound cytokines. New Biol. 1992 Feb;4(2):147–156. [PubMed] [Google Scholar]
  9. Haas G. J., Wooding-Scott M., Binkley P. F., Myerowitz P. D., Kelley R., Cody R. J. Effects of successful cardiac transplantation on plasma endothelin. Am J Cardiol. 1993 Jan 15;71(2):237–240. doi: 10.1016/0002-9149(93)90744-w. [DOI] [PubMed] [Google Scholar]
  10. Hahn A. W., Resink T. J., Scott-Burden T., Powell J., Dohi Y., Bühler F. R. Stimulation of endothelin mRNA and secretion in rat vascular smooth muscle cells: a novel autocrine function. Cell Regul. 1990 Aug;1(9):649–659. doi: 10.1091/mbc.1.9.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Halloran P. F., Broski A. P., Batiuk T. D., Madrenas J. The molecular immunology of acute rejection: an overview. Transpl Immunol. 1993;1(1):3–27. doi: 10.1016/0966-3274(93)90055-d. [DOI] [PubMed] [Google Scholar]
  12. Hancock W. W., Adams D. H., Wyner L. R., Sayegh M. H., Karnovsky M. J. CD4+ mononuclear cells induce cytokine expression, vascular smooth muscle cell proliferation, and arterial occlusion after endothelial injury. Am J Pathol. 1994 Nov;145(5):1008–1014. [PMC free article] [PubMed] [Google Scholar]
  13. Hancock W. W., Khoury S. J., Carpenter C. B., Sayegh M. H. Differential effects of oral versus intrathymic administration of polymorphic major histocompatibility complex class II peptides on mononuclear and endothelial cell activation and cytokine expression during a delayed-type hypersensitivity response. Am J Pathol. 1994 Jun;144(6):1149–1158. [PMC free article] [PubMed] [Google Scholar]
  14. Hancock W. W., Sayegh M. H., Sablinski T., Kut J. P., Kupiec-Weglinski J. W., Milford E. L. Blocking of mononuclear cell accumulation, cytokine production, and endothelial activation within rat cardiac allografts by CD4 monoclonal antibody therapy. Transplantation. 1992 Jun;53(6):1276–1280. doi: 10.1097/00007890-199206000-00022. [DOI] [PubMed] [Google Scholar]
  15. Hirata Y., Takagi Y., Fukuda Y., Marumo F. Endothelin is a potent mitogen for rat vascular smooth muscle cells. Atherosclerosis. 1989 Aug;78(2-3):225–228. doi: 10.1016/0021-9150(89)90227-x. [DOI] [PubMed] [Google Scholar]
  16. Kanno K., Hirata Y., Numano F., Emori T., Ohta K., Shichiri M., Marumo F. Endothelin-1 and vasculitis. JAMA. 1990 Dec 12;264(22):2868–2868. [PubMed] [Google Scholar]
  17. Komuro I., Kurihara H., Sugiyama T., Yoshizumi M., Takaku F., Yazaki Y. Endothelin stimulates c-fos and c-myc expression and proliferation of vascular smooth muscle cells. FEBS Lett. 1988 Oct 10;238(2):249–252. doi: 10.1016/0014-5793(88)80489-7. [DOI] [PubMed] [Google Scholar]
  18. Lee M. E., Temizer D. H., Clifford J. A., Quertermous T. Cloning of the GATA-binding protein that regulates endothelin-1 gene expression in endothelial cells. J Biol Chem. 1991 Aug 25;266(24):16188–16192. [PubMed] [Google Scholar]
  19. Lerman A., Edwards B. S., Hallett J. W., Heublein D. M., Sandberg S. M., Burnett J. C., Jr Circulating and tissue endothelin immunoreactivity in advanced atherosclerosis. N Engl J Med. 1991 Oct 3;325(14):997–1001. doi: 10.1056/NEJM199110033251404. [DOI] [PubMed] [Google Scholar]
  20. Lerman A., Kubo S. H., Tschumperlin L. K., Burnett J. C., Jr Plasma endothelin concentrations in humans with end-stage heart failure and after heart transplantation. J Am Coll Cardiol. 1992 Oct;20(4):849–853. doi: 10.1016/0735-1097(92)90183-n. [DOI] [PubMed] [Google Scholar]
  21. Lüscher T. F. Endothelial control of vascular tone and growth. Clin Exp Hypertens A. 1990;12(5):897–902. doi: 10.3109/10641969009073507. [DOI] [PubMed] [Google Scholar]
  22. Marsden P. A., Brenner B. M. Transcriptional regulation of the endothelin-1 gene by TNF-alpha. Am J Physiol. 1992 Apr;262(4 Pt 1):C854–C861. doi: 10.1152/ajpcell.1992.262.4.C854. [DOI] [PubMed] [Google Scholar]
  23. Martin-Nizard F., Houssaini H. S., Lestavel-Delattre S., Duriez P., Fruchart J. C. Modified low density lipoproteins activate human macrophages to secrete immunoreactive endothelin. FEBS Lett. 1991 Nov 18;293(1-2):127–130. doi: 10.1016/0014-5793(91)81167-7. [DOI] [PubMed] [Google Scholar]
  24. Ohta K., Hirata Y., Imai T., Kanno K., Emori T., Shichiri M., Marumo F. Cytokine-induced release of endothelin-1 from porcine renal epithelial cell line. Biochem Biophys Res Commun. 1990 Jun 15;169(2):578–584. doi: 10.1016/0006-291x(90)90370-3. [DOI] [PubMed] [Google Scholar]
  25. Perico N., Remuzzi G. Role of endothelin in glomerular injury. Kidney Int Suppl. 1993 Jan;39:S76–S80. [PubMed] [Google Scholar]
  26. Russell M. E., Adams D. H., Wyner L. R., Yamashita Y., Halnon N. J., Karnovsky M. J. Early and persistent induction of monocyte chemoattractant protein 1 in rat cardiac allografts. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6086–6090. doi: 10.1073/pnas.90.13.6086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Russell M. E., Utans U., Wallace A. F., Liang P., Arceci R. J., Karnovsky M. J., Wyner L. R., Yamashita Y., Tarn C. Identification and upregulation of galactose/N-acetylgalactosamine macrophage lectin in rat cardiac allografts with arteriosclerosis. J Clin Invest. 1994 Aug;94(2):722–730. doi: 10.1172/JCI117391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Russell M. E., Wallace A. F., Hancock W. W., Sayegh M. H., Adams D. H., Sibinga N. E., Wyner L. R., Karnovsky M. J. Upregulation of cytokines associated with macrophage activation in the Lewis-to-F344 rat transplantation model of chronic cardiac rejection. Transplantation. 1995 Feb 27;59(4):572–578. [PubMed] [Google Scholar]
  29. Sakurai T., Yanagisawa M., Inoue A., Ryan U. S., Kimura S., Mitsui Y., Goto K., Masaki T. cDNA cloning, sequence analysis and tissue distribution of rat preproendothelin-1 mRNA. Biochem Biophys Res Commun. 1991 Feb 28;175(1):44–47. doi: 10.1016/s0006-291x(05)81197-0. [DOI] [PubMed] [Google Scholar]
  30. Sayegh M. H., Sablinski T., Tanaka K., Kut J. P., Kwok C. A., Tilney N. L., Kupiec-Weglinski J. W., Milford E. L. Effects of BWH-4 anti-CD4 monoclonal antibody on rat vascularized cardiac allografts before and after engraftment. Transplantation. 1991 Feb;51(2):296–299. doi: 10.1097/00007890-199102000-00003. [DOI] [PubMed] [Google Scholar]
  31. Simonson M. S., Dunn M. J. Endothelin peptides: a possible role in glomerular inflammation. Lab Invest. 1991 Jan;64(1):1–4. [PubMed] [Google Scholar]
  32. Takagi Y., Fukase M., Takata S., Yoshimi H., Tokunaga O., Fujita T. Autocrine effect of endothelin on DNA synthesis in human vascular endothelial cells. Biochem Biophys Res Commun. 1990 Apr 30;168(2):537–543. doi: 10.1016/0006-291x(90)92354-3. [DOI] [PubMed] [Google Scholar]
  33. Wagner O. F., Christ G., Wojta J., Vierhapper H., Parzer S., Nowotny P. J., Schneider B., Waldhäusl W., Binder B. R. Polar secretion of endothelin-1 by cultured endothelial cells. J Biol Chem. 1992 Aug 15;267(23):16066–16068. [PubMed] [Google Scholar]
  34. Watschinger B., Vychytil A., Attar M., Wagner D., Schuller M., Hartter E., Ulrich W. Pattern of endothelin immunostaining during rejection episodes after kidney transplantation. Clin Nephrol. 1994 Feb;41(2):86–93. [PubMed] [Google Scholar]
  35. Watschinger B., Vychytil A., Schuller M., Hartter E., Traindl O., Pohanka E., Ulrich W., Kovarik J. The pathophysiologic role of endothelin in acute vascular rejection after renal transplantation. Transplantation. 1991 Oct;52(4):743–746. doi: 10.1097/00007890-199110000-00035. [DOI] [PubMed] [Google Scholar]
  36. Winkles J. A., Alberts G. F., Brogi E., Libby P. Endothelin-1 and endothelin receptor mRNA expression in normal and atherosclerotic human arteries. Biochem Biophys Res Commun. 1993 Mar 31;191(3):1081–1088. doi: 10.1006/bbrc.1993.1327. [DOI] [PubMed] [Google Scholar]
  37. Yamakado M., Hirata Y., Matsuoka H., Sugimoto T. Pathophysiological role of endothelin in renal transplantation. J Cardiovasc Pharmacol. 1991;17 (Suppl 7):S477–S479. doi: 10.1097/00005344-199100177-00137. [DOI] [PubMed] [Google Scholar]
  38. 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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