Troglitazone and rosiglitazone inhibit the low density lipoprotein-induced vascular smooth muscle cell growth

Summary:

Troglitazone (TRO) and rosiglitazone (RSG) belong to the thiazolidinedione class (insulin-sensitizing agents) and exert many of their metabolic effects as peroxisome proliferator-activated receptor γ (PPAR γ) ligands. In the present study we examined the effects of TRO and RSG on LDL-induced VSMC growth. Pretreatment of VSMC with 1 μM TRO or 0.1 μM RSG completely blocked the LDL-induced cell proliferation as measured by [³H]thymidine incorporation into DNA and by determination of the cell number. We then examined with Western blotting whether these growth suppressing effects are mediated through the mitogen-activated protein kinase (MAPK) pathway, a common signaling pathway activated by growth factors. TRO and RSG had no effect on the LDL-induced stimulation of the MAP kinases ERK1/2, p38 and SAP/JNK. We conclude that thiazolidinediones are potent inhibitors of LDL-induced VSMC growth acting downstream of the cytoplasmic activation of MAPK.

Abbreviations: bFGF basic fibroblast growth factor; DMSO dimethyl sulfoxide; EGF epidermal growth factor; ERK1/2 extracellular signal-response kinases (also termed p44^mapk/p42^mapk); SAPK/JNK stress-activated protein kinase/c-Jun N-terminal kinases; LDL low density lipoprotein; MAPK mitogen-activated protein kinase; MEK ERK kinase; PDVF polyvinylidene difluoride; PPARγ peroxisome proliferator-activated receptor γ; RSG rosiglitazone; TRO troglitazone; VSMC vascular smooth muscle cell(s); RXR retinoid X receptor.

Troglitazone - rosiglitazone - oral antidiabetic drugs - low density lipoprotein - MAP kinase

References

• 1 Asano M, Nakajima T, Iwasawa K, Morita T, Nakamura F, Imuta H, Chisaki K, Yamada N, Omata M, Okuda Y. Troglitazone and pioglitazone attenuate agonist-dependent Ca²⁺ mobilization and cell proliferation in vascular smooth muscle cells.  Br J Pharmacol. 128 673-683 1999; 
• 2 Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.  Anal Biochem. 72 248-254 1976; 
• 3 Chamley J H, Campbell G R, Ross R. The smooth muscle cell in culture.  Physiol Rev. 39 1-61 1979; 
• 4 Davis R J. The mitogen-activated protein kinase signal transduction pathway.  J Biol Chem. 268 14553-14556 1993; 
• 5 Dérijard B, Hibi M, Wu I H, Barrett T, Su B, Deng T, Karin M, Davis R J. JNK1: a protein kinase stimulated by UV light and Ha-Ras that binds and phosphorylates the c-Jun activation domain.  Cell. 76 1025-1037 1994; 
• 6 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. 265 R726-R732 1993; 
• 7 Fanger G R, Gerwins P, Widmann C, Jarpe M B, Johnson G L. MEKKs, GCKs, MLKs, PAKs, TAKs and tpls: upstream regulators of the c-Jun amino-terminal kinases?.  Curr Opin Genet Dev. 7 67-74 1997; 
• 8 Force T, Bonventre J V. Growth factors and mitogen-activated protein kinases.  Hypertension. 31 152-161 1998; 
• 9 Gille H, Kortenjann M, Strahl T, Shaw P E. Phosphorylation-dependent formation of a quaternary complex at the c-fos SRE.  Mol Cell Biol. 16 1094-1102 1996; 
• 10 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. 14 951-962 1995; 
• 11 Goetze S, Xi X P, Kawano H, Gotlibowski T, Fleck E, Hsueh W A, Law R E. PPARγ-ligands inhibit migration mediated by multiple chemoattractants in vascular smooth muscle cells.  J Cardiovasc Pharmacol. 33 798-806 1999; 
• 12 Han J, Lee J D, Bibbs L, Ulevitch R J. A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells.  Science. 265 808-811 1994; 
• 13 Iijima K, Yoshizumi M, Ako J, Eto M, Kim S, Hashimoto M, Sugimoto N, Liang Y Q, Sudoh N, Toba K, Ouchi Y. Expression of peroxisome proliferator-activated receptor gamma (PPARgamma) in rat aortic smooth muscle cells.  Biochemical Biophysical Research Community. 247 353-356 1998; 
• 14 Jing Q, Xin S M, Cheng Z J, Zhang W B, Zhang R, Qin Y W, Pei G. Activation of p38 mitogen-activated protein kinase by oxidized LDL in vascular smooth muscle cells: mediation via pertussis toxin-sensitive G proteins and association with oxidized LDL-induced cytotoxity.  Circ Res. 84 831-839 1999; 
• 15 Knock G A, Mishra S K, Aaronson P I. Differential effects of insulin-sensitizers troglitazone and rosiglitazone on ion currents in rat vascular myocytes.  Eur J Pharmacol. 368 103-109 1999; 
• 16 Ko Y, Totzke G, Seewald S, Schmitz U, Schiermeyer B, Meyer zu Brickwedde M, Vetter H, Sachinidis A. Native low-density lipoprotein (LDL) induces the expression of the early-growth response gene-1 in human umbilical arterial endothelial cells.  Eur J Cell Biol. 68 306-312 1995; 
• 17 Law R E, Goetze S, Xi X P, Jackson S, Kawano Y, Demer L, Fishbein M C, Meehan W P, Hsueh W A. Expression and function of PPARgamma in rat and human vascular smooth muscle cells.  Circulation. 101 1311-1318 2000; 
• 18 Law R E, Meehan W P, Xi X P, Graf K, Wuthrich D A, Coats W, Faxon D, Hsueh W A. Troglitazone inhibits vascular smooth muscle cell growth and intimal hyperplasia.  J Clin Invest. 98 1897-1905 1996; 
• 19 Lehmann J M, Moore L B, Smith-Oliver T A, Wilkison W O, Willson T M, Kliewer S A. An antidiabetic thiazolidinedione is a high affinity ligand for peroxisome proliferator-activated receptor gamma (PPARgamma).  J Biol Chem. 270 12953-12956 1995; 
• 20 Libby P, Miao P, Ordovas J M, Schaefer E J. Lipoproteins increase growth and mitogen stimulated arterial smooth muscle cell.  Journal of Cell Physiology. 124 1-8 1985; 
• 21 Marshall C J. Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation.  Cell. 80 179-185 1995; 
• 22 Marx N, Schoenbeck U, Lazar M A, Libby P, Plutzky J. Peroxisome proliferator-activated receptor gamma activators inhibit gene expression and migration in human vascular smooth muscle cells.  Circ Res. 83 1097-1103 1998; 
• 23 Metzler B, Hu Y, Dietrich H, Xu Q. Increased expression and acticvation of stress-activated protein kinases/c-Jun NH (2)-terminal protein kinases in atherosclerotic lesions coincide with p53.  Am J Pathol. 156 1875-1876 2000; 
• 24 Minamikawa J, Tanaka S, Yamauchi M, Inoue D, Koshiyama H. Potent inhibitory effect of troglitazone on carotid arterial wall thickness in type 2 diabetes.  J Clin Endocrinol Metab. 83 1818-1820 1998; 
• 25 Nakajima T, Iwasawa K, Oonuma H, Imuta H, Hazama H, Asano M, Morita T, Nakamura F, Suzuki J, Suzuki S, Kawakami Y, Omata M, Okuda Y. Troglitazone inhibits voltage-dependent calcium currents in guinea pig cardiac myocytes.  Circulation. 99 2942-2950 1999; 
• 26 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 E J M. 331 1188-1193 1994; 
• 27 Ogihara T, Rakugi H, Ikegami H, Mikami H, Masuo K. Enhancement of insulin sensitivity by troglitazone lowers blood pressure in diabetic hypertensives.  Am J Hypertens. 8 316-320 1995; 
• 28 Redgrave T G, Roberts D C, West C E. Separation of plasma lipoproteins by density-gradient ultracentrifugation.  Anal Biochem. 65 42-49 1975; 
• 29 Ricote M, Li A C, Willson T M, Kelly C J, Glass C K. The peroxisome proliferator-activated receptor-gamma is a negative regulator of macrophage activation.  Nature. 391 79-82 1998; 
• 30 Ross R. The pathogenesis of atherosclerosis: a perspective for the 1990s.  Nature. 362 801-809 1993; 
• 31 Sachinidis A, Flesch M, Ko Y, Schrör K, Böhm M, Düsing R, Vetter H. Thromboxane A2 and vascular smooth muscle cell proliferation.  Hypertension. 26 771-780 1995; 
• 32 Sachinidis A, Ko Y, Wieczorec A, Weisser B, Locher R, Vetter W, Vetter H. Lipoproteins induce expression of the early growth response gene-1 in vascular smooth muscle cells from rat.  Biochem Biophys Res Commun. 192 794-799 1993; 
• 33 Sachinidis A, Liu M, Weber A A, Seul C, Harth V, Seewald S, Ko Y, Vetter H. Cholesterol enhances platelet-derived growth factor-BB-induced [Ca2+]i and DNA synthesis in rat aortic smooth muscle cells.  Hypertension. 29 326-333 1997a; 
• 34 Sachinidis A, Mengden T, Locher R, Brunner C, Vetter W. Novel cellular activities for low-density lipoprotein in vascular smooth muscle cells.  Hypertension (Dallas). 15 704-711 1990; 
• 35 Sachinidis A, Seewald S, Epping P, Seul C, Ko Y, Vetter H. The growth-promoting effect of low-density lipoprotein may be mediated by a pertussis toxin-sensitive mitogen-activated protein kinase pathway.  Mol Pharmacol. 52 389-397 1997b; 
• 36 Schoonjans K, Martin G, Staels B, Auwerx J. Peroxisome proliferator-activated receptors, orphans with ligands and functions.  Curr Opin Lipidol. 8 159-166 1997; 
• 37 Schwartz S, Raskin P, Fonseca V, Graveline J F. Effect of troglitazone in insulin-treated patients with type II diabetes mellitus. Troglitazone and Exogenous Insulin Study Group.  N E J M. 338 861-866 1998; 
• 38 Schwarz S M, Campbell G R, Campbell J H. Replication of smooth muscle cells in vascular disease.  Circ Res. 58 427-444 1986; 
• 39 Seewald C, Nickeng G, Ko Y, Vetter H, Sachinidis A. Low density lipoprotein enhances the thrombin-induced growth of vascular smooth muscle cells.  Cardiovasc Res. 36 92-100 1997; 
• 40 Shinohara E, Kihara S, Ouchi N, Funahashi T, Nakamura T, Yamashita S, Kameda-Takemura K, Matsuzawa Y. Troglitazone suppresses intimal formation following balloon injury in insulin-resistant Zucker fatty rats.  Atherosclerosis. 136 275-279 1998; 
• 41 Spiegelman B M. PPAR-gamma: adipogenic regulator and thiazolidinedione receptor.  Diabetes. 47 507-514 1998; 
• 42 Willson T M, Cobb J E, Cowan D J, Wiethe R W, Correa I D, Prakash S R, Beck K B, Moore L B, Kliewer S A, Lehman J M. The structure-activity relationship between peroxisome proliferator-activated receptor γ agonism and the antihyperglycemic activity of thiazolidinediones.  Journal of Medical Chemistry. 39 665-668 1996; 
• 43 Yang C M, Chiu C T, Wang C C, Chien C S, Hsiao L D, Lin C C, Tu M T, Pan S L. Activation of mitogen-activated protein kinase by oxidized low-density lipoprotein in canine cultured vascular smooth muscle cells.  Cell Signal. 12 205-214 2000; 
• 44 Yasunari K, Kohno M, Kano H, Yokokawa K, Minami M, Yoshikawa J. Mechanisms of action of troglitazone in the prevention of high glucose-induced migration and proliferation of cultured coronary smooth muscle cell.  Circ Res. 81 953-962 1997;