Skip to main content
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1995 Feb;95(2):651–657. doi: 10.1172/JCI117710

The angiotensin AT2-receptor mediates inhibition of cell proliferation in coronary endothelial cells.

M Stoll 1, U M Steckelings 1, M Paul 1, S P Bottari 1, R Metzger 1, T Unger 1
PMCID: PMC295531  PMID: 7860748

Abstract

Angiotensin II (ANG II) is known to be a potent growth promoting factor for vascular smooth muscle cells and fibroblasts but little is known about its influence on growth in endothelial cells. We studied the effects of ANG II on endothelial growth and the role of the angiotensin receptor subtypes involved. Proliferation of rat coronary endothelial cells (CEC) and rat vascular smooth muscle cells (VSMC) was determined by [3H]thymidine incorporation, the MTT-test and by directly counting cells in a coulter counter. Angiotensin AT1- and AT2-receptors were demonstrated by binding studies and by the presence of their respective mRNA through reverse transcription polymerase chain reaction (RT-PCR). In contrast to VSMC, which in culture only express the AT1-receptor, CEC express both, AT1- and AT2-receptors simultaneously up to the third passage. Whereas ANG II stimulated growth of quiescent VSMC, an effect abolished by pretreatment with the AT1-receptor antagonist, losartan, ANG II did not induce proliferation in quiescent CEC. However, after pretreatment of quiescent endothelial cells (< passage 4) with the AT2-receptor antagonist, PD 123177, ANG II induced proliferation. This effect was reversed by additional pretreatment with losartan. ANG II significantly inhibited the proliferation of bFGF-stimulated CEC in a dose-dependent manner by maximally 50%. This effect was prevented by PD 123177 while losartan was ineffective. The AT2-receptor agonist, CGP 42112, mimicked the antiproliferative actions of ANG II, confirming the specificity of the effect. Our results show that the growth modulating actions of ANG II depend on the type of angiotensin receptor present on a given cell. In coronary endothelial cells, the antiproliferative actions of the AT2-receptor offset the growth promoting effects mediated by the AT1-receptor.

Full text

PDF
651

Images in this article

Selected References

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

  1. Aceto J. F., Baker K. M. [Sar1]angiotensin II receptor-mediated stimulation of protein synthesis in chick heart cells. Am J Physiol. 1990 Mar;258(3 Pt 2):H806–H813. doi: 10.1152/ajpheart.1990.258.3.H806. [DOI] [PubMed] [Google Scholar]
  2. Baird A., Durkin T. Inhibition of endothelial cell proliferation by type beta-transforming growth factor: interactions with acidic and basic fibroblast growth factors. Biochem Biophys Res Commun. 1986 Jul 16;138(1):476–482. doi: 10.1016/0006-291x(86)90305-0. [DOI] [PubMed] [Google Scholar]
  3. Berk B. C., Vekshtein V., Gordon H. M., Tsuda T. Angiotensin II-stimulated protein synthesis in cultured vascular smooth muscle cells. Hypertension. 1989 Apr;13(4):305–314. doi: 10.1161/01.hyp.13.4.305. [DOI] [PubMed] [Google Scholar]
  4. Bottari S. P., King I. N., Reichlin S., Dahlstroem I., Lydon N., de Gasparo M. The angiotensin AT2 receptor stimulates protein tyrosine phosphatase activity and mediates inhibition of particulate guanylate cyclase. Biochem Biophys Res Commun. 1992 Feb 28;183(1):206–211. doi: 10.1016/0006-291x(92)91629-5. [DOI] [PubMed] [Google Scholar]
  5. Bottari S. P., Taylor V., King I. N., Bogdal Y., Whitebread S., de Gasparo M. Angiotensin II AT2 receptors do not interact with guanine nucleotide binding proteins. Eur J Pharmacol. 1991 Jun 19;207(2):157–163. doi: 10.1016/0922-4106(91)90091-u. [DOI] [PubMed] [Google Scholar]
  6. Brechler V., Jones P. W., Levens N. R., de Gasparo M., Bottari S. P. Agonistic and antagonistic properties of angiotensin analogs at the AT2 receptor in PC12W cells. Regul Pept. 1993 Mar 19;44(2):207–213. doi: 10.1016/0167-0115(93)90244-3. [DOI] [PubMed] [Google Scholar]
  7. Brechler V., Levens N. R., De Gasparo M., Bottari S. P. Angiotensin AT2 receptor mediated inhibition of particulate guanylate cyclase: a link with protein tyrosine phosphatase stimulation? Receptors Channels. 1994;2(2):79–87. [PubMed] [Google Scholar]
  8. Brechler V., Reichlin S., De Gasparo M., Bottari S. P. Angiotensin II stimulates protein tyrosine phosphatase activity through a G-protein independent mechanism. Receptors Channels. 1994;2(2):89–98. [PubMed] [Google Scholar]
  9. Buisson B., Bottari S. P., de Gasparo M., Gallo-Payet N., Payet M. D. The angiotensin AT2 receptor modulates T-type calcium current in non-differentiated NG108-15 cells. FEBS Lett. 1992 Sep 7;309(2):161–164. doi: 10.1016/0014-5793(92)81086-2. [DOI] [PubMed] [Google Scholar]
  10. Campbell-Boswell M., Robertson A. L., Jr Effects of angiotensin II and vasopressin on human smooth muscle cells in vitro. Exp Mol Pathol. 1981 Oct;35(2):265–276. doi: 10.1016/0014-4800(81)90066-6. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Criscione L., Thomann H., Whitebread S., de Gasparo M., Bühlmayer P., Herold P., Ostermayer F., Kamber B. Binding characteristics and vascular effects of various angiotensin II antagonists. J Cardiovasc Pharmacol. 1990;16 (Suppl 4):S56–S59. doi: 10.1097/00005344-199016004-00012. [DOI] [PubMed] [Google Scholar]
  13. Fernandez L. A., Twickler J., Mead A. Neovascularization produced by angiotensin II. J Lab Clin Med. 1985 Feb;105(2):141–145. [PubMed] [Google Scholar]
  14. Geisterfer A. A., Peach M. J., Owens G. K. Angiotensin II induces hypertrophy, not hyperplasia, of cultured rat aortic smooth muscle cells. Circ Res. 1988 Apr;62(4):749–756. doi: 10.1161/01.res.62.4.749. [DOI] [PubMed] [Google Scholar]
  15. Glaser B. M., D'Amore P. A., Michels R. G., Patz A., Fenselau A. Demonstration of vasoproliferative activity from mammalian retina. J Cell Biol. 1980 Feb;84(2):298–304. doi: 10.1083/jcb.84.2.298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kambayashi Y., Bardhan S., Takahashi K., Tsuzuki S., Inui H., Hamakubo T., Inagami T. Molecular cloning of a novel angiotensin II receptor isoform involved in phosphotyrosine phosphatase inhibition. J Biol Chem. 1993 Nov 25;268(33):24543–24546. [PubMed] [Google Scholar]
  17. Katz A. M. Angiotensin II: hemodynamic regulator or growth factor? J Mol Cell Cardiol. 1990 Jul;22(7):739–747. doi: 10.1016/0022-2828(90)90086-h. [DOI] [PubMed] [Google Scholar]
  18. Le Noble F. A., Schreurs N. H., van Straaten H. W., Slaaf D. W., Smits J. F., Rogg H., Struijker-Boudier H. A. Evidence for a novel angiotensin II receptor involved in angiogenesis in chick embryo chorioallantoic membrane. Am J Physiol. 1993 Feb;264(2 Pt 2):R460–R465. doi: 10.1152/ajpregu.1993.264.2.R460. [DOI] [PubMed] [Google Scholar]
  19. Lett H. President's Address: Medicine in Transition. Br Med J. 1946 Aug 3;2(4465):149–150. doi: 10.1136/bmj.2.4465.149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lyall F., Dornan E. S., McQueen J., Boswell F., Kelly M. Angiotensin II increases proto-oncogene expression and phosphoinositide turnover in vascular smooth muscle cells via the angiotensin II AT1 receptor. J Hypertens. 1992 Dec;10(12):1463–1469. doi: 10.1097/00004872-199210120-00005. [DOI] [PubMed] [Google Scholar]
  21. Lyall F., Morton J. J., Lever A. F., Cragoe E. J. Angiotensin II activates Na+-H+ exchange and stimulates growth in cultured vascular smooth muscle cells. J Hypertens Suppl. 1988 Dec;6(4):S438–S441. doi: 10.1097/00004872-198812040-00138. [DOI] [PubMed] [Google Scholar]
  22. Montesano R., Pepper M. S., Belin D., Vassalli J. D., Orci L. Induction of angiogenesis in vitro by vanadate, an inhibitor of phosphotyrosine phosphatases. J Cell Physiol. 1988 Mar;134(3):460–466. doi: 10.1002/jcp.1041340318. [DOI] [PubMed] [Google Scholar]
  23. Mukoyama M., Nakajima M., Horiuchi M., Sasamura H., Pratt R. E., Dzau V. J. Expression cloning of type 2 angiotensin II receptor reveals a unique class of seven-transmembrane receptors. J Biol Chem. 1993 Nov 25;268(33):24539–24542. [PubMed] [Google Scholar]
  24. Naftilan A. J., Pratt R. E., Eldridge C. S., Lin H. L., Dzau V. J. Angiotensin II induces c-fos expression in smooth muscle via transcriptional control. Hypertension. 1989 Jun;13(6 Pt 2):706–711. doi: 10.1161/01.hyp.13.6.706. [DOI] [PubMed] [Google Scholar]
  25. Owens G. K., Rabinovitch P. S., Schwartz S. M. Smooth muscle cell hypertrophy versus hyperplasia in hypertension. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7759–7763. doi: 10.1073/pnas.78.12.7759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Paquet J. L., Baudouin-Legros M., Brunelle G., Meyer P. Angiotensin II-induced proliferation of aortic myocytes in spontaneously hypertensive rats. J Hypertens. 1990 Jun;8(6):565–572. doi: 10.1097/00004872-199006000-00010. [DOI] [PubMed] [Google Scholar]
  27. Paul M., Wagner J., Dzau V. J. Gene expression of the renin-angiotensin system in human tissues. Quantitative analysis by the polymerase chain reaction. J Clin Invest. 1993 May;91(5):2058–2064. doi: 10.1172/JCI116428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sandberg K., Ji H., Millan M. A., Catt K. J. Amphibian myocardial angiotensin II receptors are distinct from mammalian AT1 and AT2 receptor subtypes. FEBS Lett. 1991 Jun 24;284(2):281–284. doi: 10.1016/0014-5793(91)80704-7. [DOI] [PubMed] [Google Scholar]
  29. Stallone J. N., Nishimura H., Khosla M. C. Angiotensin II vascular receptors in fowl aorta: binding specificity and modulation by divalent cations and guanine nucleotides. J Pharmacol Exp Ther. 1989 Dec;251(3):1076–1082. [PubMed] [Google Scholar]
  30. Stouffer G. A., Owens G. K. Angiotensin II-induced mitogenesis of spontaneously hypertensive rat-derived cultured smooth muscle cells is dependent on autocrine production of transforming growth factor-beta. Circ Res. 1992 Apr;70(4):820–828. doi: 10.1161/01.res.70.4.820. [DOI] [PubMed] [Google Scholar]
  31. Tada H., Shiho O., Kuroshima K., Koyama M., Tsukamoto K. An improved colorimetric assay for interleukin 2. J Immunol Methods. 1986 Nov 6;93(2):157–165. doi: 10.1016/0022-1759(86)90183-3. [DOI] [PubMed] [Google Scholar]
  32. Timmermans P. B., Wong P. C., Chiu A. T., Herblin W. F. Nonpeptide angiotensin II receptor antagonists. Trends Pharmacol Sci. 1991 Feb;12(2):55–62. doi: 10.1016/0165-6147(91)90498-h. [DOI] [PubMed] [Google Scholar]
  33. Tsutsumi K., Saavedra J. M. Characterization and development of angiotensin II receptor subtypes (AT1 and AT2) in rat brain. Am J Physiol. 1991 Jul;261(1 Pt 2):R209–R216. doi: 10.1152/ajpregu.1991.261.1.R209. [DOI] [PubMed] [Google Scholar]
  34. Tsutsumi K., Viswanathan M., Strömberg C., Saavedra J. M. Type-1 and type-2 angiotensin II receptors in fetal rat brain. Eur J Pharmacol. 1991 May 30;198(1):89–92. doi: 10.1016/0014-2999(91)90566-9. [DOI] [PubMed] [Google Scholar]
  35. Unger T., Mattfeldt T., Lamberty V., Bock P., Mall G., Linz W., Schölkens B. A., Gohlke P. Effect of early onset angiotensin converting enzyme inhibition on myocardial capillaries. Hypertension. 1992 Oct;20(4):478–482. doi: 10.1161/01.hyp.20.4.478. [DOI] [PubMed] [Google Scholar]
  36. de Gasparo M., Whitebread S., Mele M., Motani A. S., Whitcombe P. J., Ramjoué H. P., Kamber B. Biochemical characterization of two angiotensin II receptor subtypes in the rat. J Cardiovasc Pharmacol. 1990;16 (Suppl 4):S31–S35. doi: 10.1097/00005344-199016004-00008. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES