Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1996 Oct 15;98(8):1897–1905. doi: 10.1172/JCI118991

Troglitazone inhibits vascular smooth muscle cell growth and intimal hyperplasia.

R E Law 1, W P Meehan 1, X P Xi 1, K Graf 1, D A Wuthrich 1, W Coats 1, D Faxon 1, W A Hsueh 1
PMCID: PMC507630  PMID: 8878442

Abstract

Vascular smooth muscle cell (VSMC) proliferation and migration are responses to arterial injury that are highly important to the processes of restenosis and atherosclerosis. In the arterial balloon injury model in the rat, platelet-derived growth factor (PDGF) and basic fibroblast growth factor (bFGF) are induced in the vessel wall and regulate these VSMC activities. Novel insulin sensitizing agents, thiazolidinediones, have been demonstrated to inhibit insulin and epidermal growth factor-induced growth of VSMCs. We hypothesized that these agents might also inhibit the effect of PDGF and bFGF on cultured VSMCs and intimal hyperplasia in vivo. Troglitazone (1 microM), a member of the thiazolidinedione class, produced a near complete inhibition of both bFGF-induced DNA synthesis as measured by bromodeoxyuridine incorporation (6.5+/-3.9 vs. 17.6+/-4.3% cells labeled, P < 0.05) and c-fos induction. This effect was associated with an inhibition (by 73+/-4%, P < 0.01) by troglitazone of the transactivation of the serum response element, which regulates c-fos expression. Inhibition of c-fos induction by troglitazone appeared to occur via a blockade of the MAP kinase pathway at a point downstream of MAP kinase activation by MAP kinase kinase. At this dose, troglitazone also inhibited PDGF-BB-directed migration of VSMC (by 70+/-6%, P < 0.01). These in vitro effects were operative in vivo. Quantitative image analysis revealed that troglitazone-treated rats had 62% (P < 0.001) less neointima/media area ratio 14 d after balloon injury of the aorta compared with injured rats that received no troglitazone. These results suggest troglitazone is a potent inhibitor of VSMC proliferation and migration and, thus, may be a useful agent to prevent restenosis and possibly atherosclerosis.

Full Text

The Full Text of this article is available as a PDF (402.2 KB).

Selected References

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

  1. Alessi D. R., Cuenda A., Cohen P., Dudley D. T., Saltiel A. R. PD 098059 is a specific inhibitor of the activation of mitogen-activated protein kinase kinase in vitro and in vivo. J Biol Chem. 1995 Nov 17;270(46):27489–27494. doi: 10.1074/jbc.270.46.27489. [DOI] [PubMed] [Google Scholar]
  2. Banskota N. K., Taub R., Zellner K., King G. L. Insulin, insulin-like growth factor I and platelet-derived growth factor interact additively in the induction of the protooncogene c-myc and cellular proliferation in cultured bovine aortic smooth muscle cells. Mol Endocrinol. 1989 Aug;3(8):1183–1190. doi: 10.1210/mend-3-8-1183. [DOI] [PubMed] [Google Scholar]
  3. Bilato C., Pauly R. R., Melillo G., Monticone R., Gorelick-Feldman D., Gluzband Y. A., Sollott S. J., Ziman B., Lakatta E. G., Crow M. T. Intracellular signaling pathways required for rat vascular smooth muscle cell migration. Interactions between basic fibroblast growth factor and platelet-derived growth factor. J Clin Invest. 1995 Oct;96(4):1905–1915. doi: 10.1172/JCI118236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Buchanan T. A., Meehan W. P., Jeng Y. Y., Yang D., Chan T. M., Nadler J. L., Scott S., Rude R. K., Hsueh W. A. Blood pressure lowering by pioglitazone. Evidence for a direct vascular effect. J Clin Invest. 1995 Jul;96(1):354–360. doi: 10.1172/JCI118041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Casscells W., Lappi D. A., Olwin B. B., Wai C., Siegman M., Speir E. H., Sasse J., Baird A. Elimination of smooth muscle cells in experimental restenosis: targeting of fibroblast growth factor receptors. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7159–7163. doi: 10.1073/pnas.89.15.7159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chang M. W., Barr E., Lu M. M., Barton K., Leiden J. M. Adenovirus-mediated over-expression of the cyclin/cyclin-dependent kinase inhibitor, p21 inhibits vascular smooth muscle cell proliferation and neointima formation in the rat carotid artery model of balloon angioplasty. J Clin Invest. 1995 Nov;96(5):2260–2268. doi: 10.1172/JCI118281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chang M. W., Barr E., Seltzer J., Jiang Y. Q., Nabel G. J., Nabel E. G., Parmacek M. S., Leiden J. M. Cytostatic gene therapy for vascular proliferative disorders with a constitutively active form of the retinoblastoma gene product. Science. 1995 Jan 27;267(5197):518–522. doi: 10.1126/science.7824950. [DOI] [PubMed] [Google Scholar]
  8. Chen R. H., Sarnecki C., Blenis J. Nuclear localization and regulation of erk- and rsk-encoded protein kinases. Mol Cell Biol. 1992 Mar;12(3):915–927. doi: 10.1128/mcb.12.3.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chen S. J., Chen Y. F., Miller D. M., Li H., Oparil S. Mithramycin inhibits myointimal proliferation after balloon injury of the rat carotid artery in vivo. Circulation. 1994 Nov;90(5):2468–2473. doi: 10.1161/01.cir.90.5.2468. [DOI] [PubMed] [Google Scholar]
  10. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  11. Clowes A. W., Reidy M. A., Clowes M. M. Kinetics of cellular proliferation after arterial injury. I. Smooth muscle growth in the absence of endothelium. Lab Invest. 1983 Sep;49(3):327–333. [PubMed] [Google Scholar]
  12. Davis R. J. The mitogen-activated protein kinase signal transduction pathway. J Biol Chem. 1993 Jul 15;268(20):14553–14556. [PubMed] [Google Scholar]
  13. Dubey R. K., Zhang H. Y., Reddy S. R., Boegehold M. A., Kotchen T. A. Pioglitazone attenuates hypertension and inhibits growth of renal arteriolar smooth muscle in rats. Am J Physiol. 1993 Oct;265(4 Pt 2):R726–R732. doi: 10.1152/ajpregu.1993.265.4.R726. [DOI] [PubMed] [Google Scholar]
  14. Dudley D. T., Pang L., Decker S. J., Bridges A. J., Saltiel A. R. A synthetic inhibitor of the mitogen-activated protein kinase cascade. Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7686–7689. doi: 10.1073/pnas.92.17.7686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Eldershaw T. P., Rattigan S., Cawthorne M. A., Buckingham R. E., Colquhoun E. Q., Clark M. G. Treatment with the thiazolidinedione (BRL 49653) decreases insulin resistance in obese Zucker hindlimb. Horm Metab Res. 1995 Apr;27(4):169–172. doi: 10.1055/s-2007-979932. [DOI] [PubMed] [Google Scholar]
  16. Ferns G. A., Raines E. W., Sprugel K. H., Motani A. S., Reidy M. A., Ross R. Inhibition of neointimal smooth muscle accumulation after angioplasty by an antibody to PDGF. Science. 1991 Sep 6;253(5024):1129–1132. doi: 10.1126/science.1653454. [DOI] [PubMed] [Google Scholar]
  17. Fujiwara T., Yoshioka S., Yoshioka T., Ushiyama I., Horikoshi H. Characterization of new oral antidiabetic agent CS-045. Studies in KK and ob/ob mice and Zucker fatty rats. Diabetes. 1988 Nov;37(11):1549–1558. doi: 10.2337/diab.37.11.1549. [DOI] [PubMed] [Google Scholar]
  18. Geissler J. F., Traxler P., Regenass U., Murray B. J., Roesel J. L., Meyer T., McGlynn E., Storni A., Lydon N. B. Thiazolidine-diones. Biochemical and biological activity of a novel class of tyrosine protein kinase inhibitors. J Biol Chem. 1990 Dec 25;265(36):22255–22261. [PubMed] [Google Scholar]
  19. Gille H., Kortenjann M., Strahl T., Shaw P. E. Phosphorylation-dependent formation of a quaternary complex at the c-fos SRE. Mol Cell Biol. 1996 Mar;16(3):1094–1102. doi: 10.1128/mcb.16.3.1094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gille H., Kortenjann M., Thomae O., Moomaw C., Slaughter C., Cobb M. H., Shaw P. E. ERK phosphorylation potentiates Elk-1-mediated ternary complex formation and transactivation. EMBO J. 1995 Mar 1;14(5):951–962. doi: 10.1002/j.1460-2075.1995.tb07076.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gille H., Sharrocks A. D., Shaw P. E. Phosphorylation of transcription factor p62TCF by MAP kinase stimulates ternary complex formation at c-fos promoter. Nature. 1992 Jul 30;358(6385):414–417. doi: 10.1038/358414a0. [DOI] [PubMed] [Google Scholar]
  22. Gonzalez F. A., Seth A., Raden D. L., Bowman D. S., Fay F. S., Davis R. J. Serum-induced translocation of mitogen-activated protein kinase to the cell surface ruffling membrane and the nucleus. J Cell Biol. 1993 Sep;122(5):1089–1101. doi: 10.1083/jcb.122.5.1089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Grotendorst G. R., Seppä H. E., Kleinman H. K., Martin G. R. Attachment of smooth muscle cells to collagen and their migration toward platelet-derived growth factor. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3669–3672. doi: 10.1073/pnas.78.6.3669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Harpold M. M., Evans R. M., Salditt-Georgieff M., Darnell J. E. Production of mRNA in Chinese hamster cells: relationship of the rate of synthesis to the cytoplasmic concentration of nine specific mRNA sequences. Cell. 1979 Aug;17(4):1025–1035. doi: 10.1016/0092-8674(79)90341-6. [DOI] [PubMed] [Google Scholar]
  25. Iwamoto Y., Kuzuya T., Matsuda A., Awata T., Kumakura S., Inooka G., Shiraishi I. Effect of new oral antidiabetic agent CS-045 on glucose tolerance and insulin secretion in patients with NIDDM. Diabetes Care. 1991 Nov;14(11):1083–1086. doi: 10.2337/diacare.14.11.1083. [DOI] [PubMed] [Google Scholar]
  26. Iwanishi M., Kobayashi M. Effect of pioglitazone on insulin receptors of skeletal muscles from high-fat-fed rats. Metabolism. 1993 Aug;42(8):1017–1021. doi: 10.1016/0026-0495(93)90016-h. [DOI] [PubMed] [Google Scholar]
  27. Janknecht R., Cahill M. A., Nordheim A. Signal integration at the c-fos promoter. Carcinogenesis. 1995 Mar;16(3):443–450. doi: 10.1093/carcin/16.3.443. [DOI] [PubMed] [Google Scholar]
  28. Jarrett R. J., Shipley M. J. Type 2 (non-insulin-dependent) diabetes mellitus and cardiovascular disease--putative association via common antecedents; further evidence from the Whitehall Study. Diabetologia. 1988 Oct;31(10):737–740. doi: 10.1007/BF00274775. [DOI] [PubMed] [Google Scholar]
  29. Jawien A., Bowen-Pope D. F., Lindner V., Schwartz S. M., Clowes A. W. Platelet-derived growth factor promotes smooth muscle migration and intimal thickening in a rat model of balloon angioplasty. J Clin Invest. 1992 Feb;89(2):507–511. doi: 10.1172/JCI115613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kakuta T., Currier J. W., Haudenschild C. C., Ryan T. J., Faxon D. P. Differences in compensatory vessel enlargement, not intimal formation, account for restenosis after angioplasty in the hypercholesterolemic rabbit model. Circulation. 1994 Jun;89(6):2809–2815. doi: 10.1161/01.cir.89.6.2809. [DOI] [PubMed] [Google Scholar]
  31. Kobayashi M., Iwanishi M., Egawa K., Shigeta Y. Pioglitazone increases insulin sensitivity by activating insulin receptor kinase. Diabetes. 1992 Apr;41(4):476–483. doi: 10.2337/diab.41.4.476. [DOI] [PubMed] [Google Scholar]
  32. Kortenjann M., Thomae O., Shaw P. E. Inhibition of v-raf-dependent c-fos expression and transformation by a kinase-defective mutant of the mitogen-activated protein kinase Erk2. Mol Cell Biol. 1994 Jul;14(7):4815–4824. doi: 10.1128/mcb.14.7.4815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Lee M. K., Miles P. D., Khoursheed M., Gao K. M., Moossa A. R., Olefsky J. M. Metabolic effects of troglitazone on fructose-induced insulin resistance in the rat. Diabetes. 1994 Dec;43(12):1435–1439. doi: 10.2337/diab.43.12.1435. [DOI] [PubMed] [Google Scholar]
  34. Lenormand P., Sardet C., Pagès G., L'Allemain G., Brunet A., Pouysségur J. Growth factors induce nuclear translocation of MAP kinases (p42mapk and p44mapk) but not of their activator MAP kinase kinase (p45mapkk) in fibroblasts. J Cell Biol. 1993 Sep;122(5):1079–1088. doi: 10.1083/jcb.122.5.1079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Libby P., Warner S. J., Salomon R. N., Birinyi L. K. Production of platelet-derived growth factor-like mitogen by smooth-muscle cells from human atheroma. N Engl J Med. 1988 Jun 9;318(23):1493–1498. doi: 10.1056/NEJM198806093182303. [DOI] [PubMed] [Google Scholar]
  36. Lindner V., Lappi D. A., Baird A., Majack R. A., Reidy M. A. Role of basic fibroblast growth factor in vascular lesion formation. Circ Res. 1991 Jan;68(1):106–113. doi: 10.1161/01.res.68.1.106. [DOI] [PubMed] [Google Scholar]
  37. Lindner V., Reidy M. A. Proliferation of smooth muscle cells after vascular injury is inhibited by an antibody against basic fibroblast growth factor. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3739–3743. doi: 10.1073/pnas.88.9.3739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Majesky M. W., Reidy M. A., Bowen-Pope D. F., Hart C. E., Wilcox J. N., Schwartz S. M. PDGF ligand and receptor gene expression during repair of arterial injury. J Cell Biol. 1990 Nov;111(5 Pt 1):2149–2158. doi: 10.1083/jcb.111.5.2149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Miano J. M., Vlasic N., Tota R. R., Stemerman M. B. Localization of Fos and Jun proteins in rat aortic smooth muscle cells after vascular injury. Am J Pathol. 1993 Mar;142(3):715–724. [PMC free article] [PubMed] [Google Scholar]
  40. Miano J. M., Vlasic N., Tota R. R., Stemerman M. B. Smooth muscle cell immediate-early gene and growth factor activation follows vascular injury. A putative in vivo mechanism for autocrine growth. Arterioscler Thromb. 1993 Feb;13(2):211–219. doi: 10.1161/01.atv.13.2.211. [DOI] [PubMed] [Google Scholar]
  41. Morishita R., Gibbons G. H., Ellison K. E., Nakajima M., von der Leyen H., Zhang L., Kaneda Y., Ogihara T., Dzau V. J. Intimal hyperplasia after vascular injury is inhibited by antisense cdk 2 kinase oligonucleotides. J Clin Invest. 1994 Apr;93(4):1458–1464. doi: 10.1172/JCI117123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Nolan J. J., Ludvik B., Beerdsen P., Joyce M., Olefsky J. Improvement in glucose tolerance and insulin resistance in obese subjects treated with troglitazone. N Engl J Med. 1994 Nov 3;331(18):1188–1193. doi: 10.1056/NEJM199411033311803. [DOI] [PubMed] [Google Scholar]
  43. Parker T. G., Chow K. L., Schwartz R. J., Schneider M. D. Positive and negative control of the skeletal alpha-actin promoter in cardiac muscle. A proximal serum response element is sufficient for induction by basic fibroblast growth factor (FGF) but not for inhibition by acidic FGF. J Biol Chem. 1992 Feb 15;267(5):3343–3350. [PubMed] [Google Scholar]
  44. Pauly R. R., Bilato C., Sollott S. J., Monticone R., Kelly P. T., Lakatta E. G., Crow M. T. Role of calcium/calmodulin-dependent protein kinase II in the regulation of vascular smooth muscle cell migration. Circulation. 1995 Feb 15;91(4):1107–1115. doi: 10.1161/01.cir.91.4.1107. [DOI] [PubMed] [Google Scholar]
  45. Pershadsingh H. A., Szollosi J., Benson S., Hyun W. C., Feuerstein B. G., Kurtz T. W. Effects of ciglitazone on blood pressure and intracellular calcium metabolism. Hypertension. 1993 Jun;21(6 Pt 2):1020–1023. doi: 10.1161/01.hyp.21.6.1020. [DOI] [PubMed] [Google Scholar]
  46. Peters K. G., Marie J., Wilson E., Ives H. E., Escobedo J., Del Rosario M., Mirda D., Williams L. T. Point mutation of an FGF receptor abolishes phosphatidylinositol turnover and Ca2+ flux but not mitogenesis. Nature. 1992 Aug 20;358(6388):678–681. doi: 10.1038/358678a0. [DOI] [PubMed] [Google Scholar]
  47. Ross R. The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature. 1993 Apr 29;362(6423):801–809. doi: 10.1038/362801a0. [DOI] [PubMed] [Google Scholar]
  48. Sandouk T., Reda D., Hofmann C. The antidiabetic agent pioglitazone increases expression of glucose transporters in 3T3-F442A cells by increasing messenger ribonucleic acid transcript stability. Endocrinology. 1993 Jul;133(1):352–359. doi: 10.1210/endo.133.1.8319581. [DOI] [PubMed] [Google Scholar]
  49. Savage P. J. Cardiovascular complications of diabetes mellitus: what we know and what we need to know about their prevention. Ann Intern Med. 1996 Jan 1;124(1 Pt 2):123–126. doi: 10.7326/0003-4819-124-1_part_2-199601011-00008. [DOI] [PubMed] [Google Scholar]
  50. Schwartz S. M., deBlois D., O'Brien E. R. The intima. Soil for atherosclerosis and restenosis. Circ Res. 1995 Sep;77(3):445–465. doi: 10.1161/01.res.77.3.445. [DOI] [PubMed] [Google Scholar]
  51. Seger R., Krebs E. G. The MAPK signaling cascade. FASEB J. 1995 Jun;9(9):726–735. [PubMed] [Google Scholar]
  52. Simons M., Edelman E. R., Rosenberg R. D. Antisense proliferating cell nuclear antigen oligonucleotides inhibit intimal hyperplasia in a rat carotid artery injury model. J Clin Invest. 1994 Jun;93(6):2351–2356. doi: 10.1172/JCI117240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Sleigh M. J. A nonchromatographic assay for expression of the chloramphenicol acetyltransferase gene in eucaryotic cells. Anal Biochem. 1986 Jul;156(1):251–256. doi: 10.1016/0003-2697(86)90180-6. [DOI] [PubMed] [Google Scholar]
  54. Sollott S. J., Cheng L., Pauly R. R., Jenkins G. M., Monticone R. E., Kuzuya M., Froehlich J. P., Crow M. T., Lakatta E. G., Rowinsky E. K. Taxol inhibits neointimal smooth muscle cell accumulation after angioplasty in the rat. J Clin Invest. 1995 Apr;95(4):1869–1876. doi: 10.1172/JCI117867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Stout R. W. Insulin and atheroma. 20-yr perspective. Diabetes Care. 1990 Jun;13(6):631–654. doi: 10.2337/diacare.13.6.631. [DOI] [PubMed] [Google Scholar]
  56. Treisman R. Journey to the surface of the cell: Fos regulation and the SRE. EMBO J. 1995 Oct 16;14(20):4905–4913. doi: 10.1002/j.1460-2075.1995.tb00173.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Twentyman P. R., Luscombe M. A study of some variables in a tetrazolium dye (MTT) based assay for cell growth and chemosensitivity. Br J Cancer. 1987 Sep;56(3):279–285. doi: 10.1038/bjc.1987.190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Zempo N., Koyama N., Kenagy R. D., Lea H. J., Clowes A. W. Regulation of vascular smooth muscle cell migration and proliferation in vitro and in injured rat arteries by a synthetic matrix metalloproteinase inhibitor. Arterioscler Thromb Vasc Biol. 1996 Jan;16(1):28–33. doi: 10.1161/01.atv.16.1.28. [DOI] [PubMed] [Google Scholar]
  59. van de Loosdrecht A. A., Beelen R. H., Ossenkoppele G. J., Broekhoven M. G., Langenhuijsen M. M. A tetrazolium-based colorimetric MTT assay to quantitate human monocyte mediated cytotoxicity against leukemic cells from cell lines and patients with acute myeloid leukemia. J Immunol Methods. 1994 Sep 14;174(1-2):311–320. doi: 10.1016/0022-1759(94)90034-5. [DOI] [PubMed] [Google Scholar]

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

RESOURCES