Published online before print June 25, 2007, doi: 10.1161/HYPERTENSIONAHA.107.091264
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
(Hypertension. 2007;50:377.)
© 2007 American Heart Association, Inc.
Original Articles |
HMG-CoA Reductase Inhibitors Decrease Angiotensin II–Induced Vascular Fibrosis
Role of RhoA/ROCK and MAPK Pathways
From the Vascular and Renal Research Laboratory, Cellular Biology in Renal Diseases Laboratory, Fundación Jiménez Diaz, Universidad Autónoma Madrid, Spain.
Correspondence to Marta Ruiz-Ortega, Cellular Biology in Renal Diseases Laboratory, Fundación Jiménez Díaz, Avda, Reyes Católicos, 2, 28040 Madrid, Spain. E-mail mruizo{at}fjd.es
 |
Abstract |
|---|
 Top Abstract IntroductionMethodsResultsDiscussionReferences |
|---|
3-hydroxy-3-methylglutaryl (HMG)-coenzyme A (CoA) reductase inhibitors (statins) present beneficial effects in cardiovascular diseases. Angiotensin II (Ang II) contributes to cardiovascular damage through the production of profibrotic factors, such as connective tissue growth factor (CTGF). Our aim was to investigate whether HMG-CoA reductase inhibitors could modulate Ang II responses, evaluating CTGF expression and the mechanisms underlying this process. In cultured vascular smooth muscle cells (VSMCs) atorvastatin and simvastatin inhibited Ang II–induced CTGF production. The inhibitory effect of statins on CTGF upregulation was reversed by mevalonate and geranylgeranylpyrophosphate, suggesting that RhoA inhibition could be involved in this process. In VSMCs, statins inhibited Ang II–induced Rho membrane localization and activation. In these cells Ang II regulated CTGF via RhoA/Rho kinase activation, as shown by inhibition of Rho with C3 exoenzyme, RhoA dominant-negative overexpression, and Rho kinase inhibition. Furthermore, activation of p38MAPK and JNK, and redox process were also involved in Ang II–mediated CTGF upregulation, and were downregulated by statins. In rats infused with Ang II (100 ng/kg per minute) for 2 weeks, treatment with atorvastatin (5 mg/kg per day) diminished aortic CTGF and Rho activation without blood pressure modification. Rho kinase inhibition decreased CTGF upregulation in rat aorta, mimicking statin effect. CTGF is a vascular fibrosis mediator. Statins diminished extracellular matrix (ECM) overexpression caused by Ang II in vivo and in vitro. In summary, HMG-CoA reductase inhibitors inhibit several intracellular signaling systems activated by Ang II (RhoA/Rho kinase and MAPK pathways and redox process) involved in the regulation of CTGF. Our results may explain, at least in part, some beneficial effects of statins in cardiovascular diseases.
Key Words: angiotensin • RhoA • Rho kinase • extracellular matrix • HMG CoA reductase • vascular smooth muscle cells
|
Introduction |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Several clinical trials have demonstrated that 3-hydroxy-3-methylglutaryl (HMG)-coenzyme A (CoA) reductase inhibitors (statins) exert beneficial effects in patients at high cardiovascular risk.1,2 Statins possess pleiotropic effects including improvement of endothelial function, reduction of blood thrombogenicity, oxidative stress, and cell proliferation, as well as antiinflammatory and immunomodulatory properties.1 These nonlipid related actions could be explained by the inhibition of several intracellular pathways, including kinases and small G proteins and exerting antioxidant properties.1 Statins through the inhibition of isoprenoid intermediates of the cholesterol biosynthetic pathway, such as farnesylpyrophosphate (FPP) and geranylgeranylpyrophosphate (GGPP), regulate posttranslational modifications of several proteins, including small GTPases-binding proteins.1 In vascular smooth muscle cells (VSMCs), several groups have shown that statins regulate cellular location and activation of Ras and Rho.1,2
Angiotensin II (Ang II) participates in the pathogenesis of cardiovascular diseases. In hypertension, Ang II via AT1 receptors induces vascular structural changes, including cell hypertrophy, accumulation of extracellular matrix (ECM) proteins, and induction of profibrotic growth factors, such as connective tissue growth factor (CTGF).3 This growth factor also mediates cell proliferation/apoptosis, migration, cellular adhesion, wound healing, and angiogenesis.4 CTGF overexpression has been found in human atherosclerotic and myocardial lesions as well as in the aorta of Ang II–infused rats, associated with ECM accumulation.5–7 In VSMCs, CTGF acts as a downstream mediator of Ang II–induced fibrosis.7 The AT1 are G-coupled receptors that activate several intracellular signaling systems, such as protein kinases (MAPK and Rho kinase [ROCK]), small G proteins, including Ras, Rac1 and RhoA, transcription factors (AP-1, Smads), and redox process, which regulate several Ang II–mediated responses.3,8
In this study, we have investigated whether statins could directly modulate Ang II fibrotic responses in vascular tissue, studying the regulation of CTGF and the molecular mechanisms underlying this process, as the role of the activation of Rho/ROCK, MAPK, and redox process.
|
Methods |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
The materials and methods used in this study are described in the online supplement (available online at http://hyper.ahajournals.org).
|
Results |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
HMG-CoA Reductase Inhibitors Decrease CTGF Production Elicited by Ang II in Cultured VSMCs
We have previously demonstrated that in VSMCs Ang II upregulates CTGF.7 VSMCs were pretreated for 1 hour with 2 statins: atorvastatin and simvastatin (10–6 mol/L to 10–9 mol/L). Both statins diminished CTGF production elicited by Ang II after 24 hours of incubation (Figure 1 and Figure S1).
|
P<0.05 vs Ang II+atorvastatin.
The Effect of Statins Is Mediated Through the Inhibition of Isoprenoids
We tested the effect of mevalonate, the direct HMG-CoA reductase metabolite, to check whether statins effect on CTGF regulation was attributable to direct inhibition of this enzyme. In the presence of mevalonate the inhibitory effect was reversed. We further investigated the implication of isoprenoid compounds on CTGF regulation. GGPP, but not FPP, markedly reversed the induction of CTGF by Ang II (Figure 1 and Figure S1), suggesting that geranylgeranylated proteins and therefore Rho signaling participate in CTGF regulation, whereas farnesylated proteins are not implicated.
HMG-CoA Reductase Inhibitors Modulate Ang II–Induced Rho Activation in Cultured VSMCs
We next evaluated whether statins could inhibit the activation of the small G protein Rho caused by Ang II. In the quiescent state, Rho binds to GDP and resides in the cytosol. On activation, GDP-Rho is converted to GTP-Rho and translocated to the membrane. In VSMCs by indirect immunofluorescent staining, we observed a diffuse cytoplasmic RhoA staining in unstimulated cells. Treatment with Ang II for 15 minutes changed staining pattern from diffuse cytosolic to membrane localized, indicating activation of RhoA. This change in RhoA distribution was blocked by atorvastatin or simvastatin (pretreated both 2 or 24 hours), and reverted by mevalonate (Figure S2). Stimulation with Ang II for 20 minutes increased GTP-bound Rho levels, showed by pull-down assays, which were diminished by pretreatment with atorvastatin (Figure 2A). We have further evaluated the effect of statins on ROCK activation, determining phosphorylation of Myosin-binding subunit of Myosin phosphatase (MYPT1), which is a specific substrate of ROCK.9 Atorvastatin significantly diminished Ang II–induced phosphorylation of MYPT1 (Figure 2B). These data demonstrate that statins inhibit Rho activation caused by Ang II.
|
Ang II Regulates CTGF Production via RhoA/ROCK Pathways
Ang II via AT1 receptors increases CTGF through the activation of several intracellular signals, including production of reactive oxygen species (ROS), activation of kinases and transcription factors, showing differences between cell types.10–12 In VSMCs, Ang II regulates CTGF via redox process and Smad pathway.7,13 However, the involvement of different kinases, such as ROCK and MAPK, has not been evaluated.
Treatment of VSMCs with 5 µg/mL C3 exoenzyme for 48 hours, time necessary to inhibit Rho GTPase activity and completely ADP-ribosylates RhoA,14 significantly attenuated CTGF production stimulated by Ang II for 24 hour (Figure S3). Transient transfection of VSMCs with a plasmid encoding constitutively active form of RhoA increased CTGF production. Overexpression of the dominant negative RhoA blocked CTGF production caused by Ang II, although it did not modify CTGF levels on unstimulated cells. As expected, in cells transfected with empty vector, Ang II increased CTGF production (Figure S3). These findings reveal that RhoA is involved in Ang II–induced CTGF upregulation. Pretreatment of VSMCs with the selective inhibitor of the serine/threonine ROCK I and II, Y-27632 (10–5 mol/L a 10–8 mol/L), dose-dependently suppressed Ang II–induced CTGF protein production. Fasudil, another ROCK inhibitor, also abrogated CTGF overexpression caused by Ang II. ROCK inhibition diminished Ang II–induced CTGF mRNA overexpression (Figures S3 and S4). These results indicates that Ang II through RhoA/ROCK pathway increases CTGF production.
HMG-CoA Reductase Inhibition Diminishes Ang II–Induced MAPK Activation in Cultured VSMCs
The involvement of MAPKs cascade on CTGF regulation was evaluated using specific inhibitors of p38-MAPK (SB203580), extracellular signal-regulated kinase (ERK) 1/2 (PD98059), and Jun N-terminal kinase (JNK) (SP600125) cascades. Only p38 and JNK inhibitors diminished Ang II–induced CTGF production (Figure S3). In fibroblasts, inhibition of ERK1/2 and JNK, but not p38MAPK, decreased Ang II–stimulated CTGF expression,15 showing a different response depending on the cell type. These data show that in VSMCs Ang II upregulates CTGF via activation of RhoA/ROCK, p38MAPK, and JNK (Figure 3S), redox process,7 and Smad pathway.13
We next evaluated whether statins could inhibit Ang II–induced activation of these pathways (Figure 3 and Figure S5). In VSMCs, Ang II triggered phosphorylation of all 3 MAPKs, with a maximal response between 10 to 15 minutes, as described.16 Preincubation with atorvastatin inhibited Ang II–induced activation of p38MAPK, JNK, and ERK1/2, suggesting that inhibition of p38 and JNK could be involved in CTGF downregulation by statins. As we have previously found, incubation with Ang II increases Smad phosphorylation at 15 minutes, but atorvastatin had no effect (not shown), suggesting that statins did not regulate Ang II–induced Smad activation. Statins exert antioxidant properties.1 The preincubation with dyphenyleneiodonium (DPI, an inhibitor of flavoprotein-containing enzymes such as NADH/NADPH oxidase) or 4,5-dihydroxy-1,3-benzenedisulfonic (Tiron, O2– scavenger), significantly reduced Ang II–induced ERK and JNK activation, showing that JNK is a common pathway for statins and antioxidants in CTGF regulation. These antioxidants also diminished Ang II–induced phosphorylation of MYPT1 (Figure S5), suggesting that Rho/ROCK pathway is modulated by redox mechanisms.
|
HMG-CoA Reductase Inhibitors Diminish Vascular Overexpression of CTGF and ECM Proteins and Rho Activation in Ang II–Infused Rats
Ang II infusion in rats caused vascular fibrosis characterized by induction of profibrotic factors and ECM accumulation.7 In Ang II–infused rats for 2 weeks we have found aortic CTGF mRNA upregulation and protein production. By immunohistochemistry, CTGF staining was mainly located in VSMCs (Figure 4), confirming and extending our previous findings.7 Atorvastatin treatment at 5 mg/Kg/d diminished CTGF mRNA and protein expression in the aorta of Ang II–infused rats (Figure 4). We have evaluated the effect of atorvastatin in Ang II–induced fibrosis. Atorvastatin diminished type I collagen deposition and type IV procollagen gene overexpression in Ang II–infused rats (Figure 4 and Figure S6). Ang II may modulate the expression of the matrix metalloproteinases (MMPs), the main ECM degradation enzymes. Ang II–induced MMP-9 gene overexpression which was decreased by atorvastatin (Figure S6).
|
The aorta of Ang II–infused rats presented elevated Rho levels, as assessed by pull down assay using the Rho binding domain of Rhotekin, the Rho effector protein which interacts only with activated GTP-bound form of Rho. This Rho activation was suppressed by atorvastatin (Figure 4C). These data demonstrate that atorvastatin inhibits Rho activation caused by Ang II in vivo.
Treatment with atorvastatin did not diminish Ang II–induced blood pressure elevation (Ang II:149±3 mm Hg, Ang II+Atorvastatin: 150±2, P=ns versus Ang II; control: 119±1 mm Hg, n=10 to 15 animals per group), showing that atorvastatin effect on Ang II–induced vascular responses was independent of blood pressure regulation.
ROCK Inhibition Diminishes CTGF Expression in the Aorta of Ang II–Infused Rats
Treatment of Ang II–infused rats with Y-27632 diminished aortic CTGF mRNA and protein overexpression (Figure 5), showing the involvement of Rho/Rho-kinase pathway in CTGF upregulation.
|
HMG-CoA Reductase and ROCK Inhibitors Decrease Ang II–Induced ECM Upregulation in Cultured VSMCs
CTGF is a downstream mediator of Ang II–induced fibrosis,7 we have evaluated whether statins could also regulate ECM proteins (Figure S4). In VSMCs, Ang II increased the gene expression of different ECM-related proteins, including fibronectin and type IV collagen, which were significantly diminished by atorvastatin, in parallel with CTGF regulation. Some of these genes were also downregulated by ROCK inhibition. Fibronectin production caused by Ang II was also diminished by atorvastatin. Transforming growth factor (TGF)-ß mediates many of the profibrotic actions of Ang II, including CTGF upregulation.7 We have observed that in VSMCs atorvastatin and Y-27632 inhibited TGF-ß mRNA expression and release. Ang II also induced plasminogen activator inhibitor (PAI)-1 upregulation, which was diminished by atorvastatin and Y-27632, indicating a potential antithrombotic role of atorvastatin.
|
Discussion |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
In the present work we have demonstrated that the HMG-CoA reductase inhibition decreased Ang II–induced CTGF production in aorta and cultured VSMCs, showing a direct cellular effect of HMG-CoA reductase inhibitors on Ang II vascular responses. We have also found that the molecular mechanisms involved in this beneficial effect is through the inhibition of Rho activation and modulation of MAPK pathway.
Several studies, most of them done in experimental models of cardiac damage, suggested that statins could decrease hypertrophy and fibrosis.1 In a model of double transgenic rats for human renin and angiotensinogen, cerivastatin ameliorated cardiac hypertrophy and interstitial fibrosis.17 In ischemia-reperfusion injury, fluvastatin diminished cardiac myocyte hypertrophy and interstitial fibrosis.18 Simvastatin also decreased Ang II–induced cardiac myocyte hypertrophy both in vivo and in vitro.19 In a rat model of systemic infusion of Ang II we have observed that atorvastatin diminished overexpression of CTGF and ECM proteins in the aorta, showing that statins could regulate Ang II–induced vascular fibrosis. Atorvastatin did not modify Ang II–induced blood pressure elevation, indicating that its beneficial effect is independent of blood pressure regulation. Moreover, in cultured VSMCs we have observed that HMG-CoA reductase inhibition downregulated CTGF and ECM overexpression caused by Ang II, showing a direct cellular effect. The relationship between statins and CTGF has also been described in other cell types. In renal cells, statins inhibited TGF-ß–induced CTGF expression.20 In renal cells and human VSMCs, HMG-CoA reductase inhibitors diminished several ECM proteins (thrombospondin-1, collagens, biglycan, and fibronectin).20–22 TGF-ß is one of the main regulators of ECM.23 In vivo coinjection of TGF-ß and CTGF promotes a persistent skin fibrosis.24 Our data show that statins inhibit TGF-ß gene expression and release caused by Ang II, indicating that the antifibrotic effect of statins could be attributable to downregulation of both profibrotic factors, CTGF and TGF-ß. Concerning matrix turnover, statins decrease MMP and other enzymes involved in ECM degradation.18,25 We have found that Ang II–induced aortic gene MMP-9 overexpression was decreased by atorvastatin. These data show that statins modulate Ang II–mediated pathological responses, diminishing vascular fibrosis, and affording a potential explanation about the beneficial actions of these drugs in hypertension and other cardiovascular diseases.
RhoA/ROCK pathway participates in pathophysiological functions of Ang II26 such as contraction,14 cardiovascular hypertrophy,27 atherogenic gene expression,28 and migration.9 Our experiments in cultured VSMCs have demonstrated that Ang II regulated CTGF via RhoA/ROCK activation, as shown by inhibition of RhoA with C3 exoenzyme, RhoA dominant-negative overexpression, and ROCK inhibition with Y-27632 and fasudil. In rats infused with Ang II, ROCK inhibition also decreased aortic CTGF gene and protein upregulation. Several groups have presented some evidences supporting our results. Rho proteins regulate CTGF induced by TGF-ß in fibroblasts.10,20 ROCK regulates Ang II–induced gene expression of atrial natriuretic peptide, monocytic chemoattractant protein-1, and PAI-1.28–30 In hypertensive rats, the ROCK system in blood vessels is activated and contributes to the pathogenesis of hypertensive vascular disease.31,32 In rats treated with L-NAME ROCK inhibition decreased vascular inflammation, cardiac fibrosis, and glomerulosclerosis, whereas AT1 blockade diminished ROCK activation.33 In this animal model statins ameliorate vascular lesions without lipids level modification, showing therefore cholesterol-independent effects.34 In a model of atherosclerosis, gene transfer of a dominant-negative ROCK vector decreased neointimal formation.35
In cultured VSMCs, we have demonstrated that the inhibitory effect on CTGF regulation caused by the HMG-CoA reductase inhibitors atorvastatin and simvastatin was reversed by mevalonate, the precursor not only of cholesterol but also of many nonsteroidal isoprenoid compounds. The addition of the isoprenoid GGPP, but not FPP, reversed statin-induced CTGF downregulation, suggesting that geranylgeranylated proteins participate in CTGF regulation. One of the geranylgeranylated proteins is RhoA. In cultured VSMCs, Ang II rapidly increases GTP-bound Rho levels, changes RhoA localization from the cytosol to the membrane, and phosphorylates the ROCK substrate MYPT1, indicating RhoA/ROCK activation. Those processes were inhibited by pretreatment with statins, clearly demonstrating that these drugs inhibit Ang II–mediated RhoA/ROCK activation. We have also found that Ang II–infused rats presented activated Rho levels in the aorta, as observed in different tissues in experimental models of hypertension,31,33,36 which was diminished by atorvastatin. In a model of cardiac injury statins also diminished Rho membrane translocation in the heart.37 These data suggest that in vivo the beneficial effect of statins in Ang II–induced vascular fibrosis could be attributable to Rho inhibition.
Statins present pleiotropic effects and can inhibit several intracellular signals activated by Ang II, including, transcription factors, such as JAK-STAT,38 NF-
B, and AP-1,39 ROS production,40 and as we have shown here several kinases. In VSMCs, we have observed that Ang II upregulates CTGF via activation of p38MAPK, JNK, and ROCK. Those kinases were inhibited by Atorvastatin, showing a potential mechanisms involved in CTGF downregulation. Ohtsu et al9 have shown that Rho/ROCK activation by Ang II specifically mediates JNK activation leading to migration of VSMCs. In accord with these data we have found that ROCK and JNK mediates Ang II–induced CTGF upregulation, and those processes were inhibited by atorvastatin. Antioxidants significantly reduced JNK and ROCK activation and CTGF upregulation caused by Ang II, showing share targets with statins. Our data suggest that statins inhibit some Ang II signaling systems, including production of ROS and activation of ROCK and JNK, that participates in CTGF production (Figure S7).
Perspectives
We have found that statins modulate some Ang II vascular responses through the inhibition of intracellular signaling systems, including RhoA/ROCK and MAPK pathways. These findings support the hypothesis that some beneficial effects of HMG-CoA reductase inhibitors described in normolipidemic hypertensive patients could be attributable to direct cellular effects, beyond cholesterol lowering effects. In addition, activation of RhoA/ROCK pathway participates in vascular damage in several experimental models,31,33,36 as we have described in Ang II–infused rats. These data support the idea that treatments that inhibit this pathway, including statins, could be a promising approach as therapeutic strategies in cardiovascular diseases.41
|
Acknowledgments |
|---|
Authors thank Mar Gonzalez Garcia-Parreño for her technical help with the confocal microscopy.
Sources of Funding
This work has been supported by grants from Fondo de Investigación Sanitaria (FIS) (PI0205513, PI020822, CP04/00060), Ministerio de Educación y Clencia, SAF 2005-03378 and SAF 2004/0610, Fundación Mapfre Medicina, and Instituto de Salud Carlos III, RECAVA (RD06/0014/0035) and ISCIII-RETIC RD06/0004. M.R., E.S.-L., and J.R.-V. are fellows of FIS.
Disclosures
None.
Received April 11, 2007; first decision April 29, 2007; accepted May 31, 2007.
|
References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
- Liao JK. Effects of statins on 3-hydroxy-3-methylglutaryl coenzyme a reductase inhibition beyond low-density lipoprotein cholesterol. Am J Cardiol. 2005; 96: 24F–33F.[Medline] [Order article via Infotrieve]
- Guijarro C, Blanco-Colio LM, Ortego M, Alonso C, Ortiz A, Plaza JJ, Diaz C, Hernandez G, Egido J. 3-Hydroxy-3-methylglutaryl coenzyme a reductase and isoprenylation inhibitors induce apoptosis of vascular smooth muscle cells in culture. Circ Res. 1998; 83: 490–500.
[Abstract/Free Full Text] - Ruiz-Ortega M, Ruperez M, Esteban V, Egido J. Molecular mechanisms of angiotensin II-induced vascular injury. Curr Hypertens Rep. 2003; 5: 73–79.[Medline] [Order article via Infotrieve]
- Perbal B. CCN proteins: multifunctional signalling regulators. Lancet. 2004; 363: 62–64.[CrossRef][Medline] [Order article via Infotrieve]
- Oemar BS, Werner A, Garnier JM, Do DD, Godoy N, Nauck M, Marz W, Rupp J, Pech M, Luscher TF. Human connective tissue growth factor is expressed in advanced atherosclerotic lesions. Circulation. 1997; 95: 831–839.
[Abstract/Free Full Text] - Chen MM, Lam A, Abraham JA, Schreiner GF, Joly AH. CTGF expression is induced by TGF- beta in cardiac fibroblasts and cardiac myocytes: a potential role in heart fibrosis. J Mol Cell Cardiol. 2000; 32: 1805–1819.[CrossRef][Medline] [Order article via Infotrieve]
- Ruperez M, Lorenzo O, Blanco-Colio LM, Esteban V, Egido J, Ruiz-Ortega M. Connective tissue growth factor is a mediator of angiotensin II-induced fibrosis. Circulation. 2003; 108: 1499–1505.
[Abstract/Free Full Text] - Ruiz-Ortega M, Ortiz A. Angiotensin II and reactive oxygen species. Antioxid Redox Signal. 2005; 7: 1258–1260.[CrossRef][Medline] [Order article via Infotrieve]
- Ohtsu H, Mifune M, Frank GD, Saito S, Inagami T, Kim-Mitsuyama S, Takuwa Y, Sasaki T, Rothstein JD, Suzuki H, Nakashima H, Woolfolk EA, Motley ED, Eguchi S. Signal-crosstalk between Rho/ROCK and c-Jun NH2-terminal kinase mediates migration of vascular smooth muscle cells stimulated by angiotensin II. Arterioscler Thromb Vasc Biol. 2005; 25: 1831–1836.
[Abstract/Free Full Text] - Iwanciw D, Rehm M, Porst M, Goppelt-Struebe M. Induction of connective tissue growth factor by angiotensin II: integration of signaling pathways. Arterioscler Thromb Vasc Biol. 2003; 23: 1782–1787.
[Abstract/Free Full Text] - He Z, Way KJ, Arikawa E, Chou E, Opland DM, Clermont A, Isshiki K, Ma RC, Scott JA, Schoen FJ, Feener EP, King GL. Differential regulation of angiotensin II-induced expression of connective tissue growth factor by protein kinase C isoforms in the myocardium. J Biol Chem. 2005; 280: 15719–15726.
[Abstract/Free Full Text] - Ruperez M, Sanchez-Lopez E, Blanco-Colio LM, Esteban V, Rodriguez-Vita J, Plaza JJ, Egido J, Ruiz-Ortega M. The Rho-kinase pathway regulates angiotensin II-induced renal damage. Kidney Int Suppl. 2005; S39–S45.
- Rodriguez-Vita J, Sanchez-Lopez E, Esteban V, Ruperez M, Egido J, Ruiz-Ortega M. Angiotensin II activates the Smad pathway in vascular smooth muscle cells by a transforming growth factor-beta-independent mechanism. Circulation. 2005; 111: 2509–2517.
[Abstract/Free Full Text] - Aoki H, Izumo S, Sadoshima J. Angiotensin II activates RhoA in cardiac myocytes: a critical role of RhoA in angiotensin II-induced premyofibril formation. Circ Res. 1998; 82: 666–676.
[Abstract/Free Full Text] - Liu B, Yu J, Taylor L, Zhou X, Polgar P. Microarray and phosphokinase screenings leading to studies on ERK and JNK regulation of connective tissue growth factor expression by angiotensin II 1a and bradykinin B2 receptors in Rat1 fibroblasts. J Cell Biochem. 2006; 97: 1104–1120.[CrossRef][Medline] [Order article via Infotrieve]
- Kim S, Iwao H. Stress and vascular responses: mitogen-activated protein kinases and activator protein-1 as promising therapeutic targets of vascular remodeling. J Pharmacol Sci. 2003; 91: 177–181.[CrossRef][Medline] [Order article via Infotrieve]
- Dechend R, Fiebeler A, Park JK, Muller DN, Theuer J, Mervaala E, Bieringer M, Gulba D, Dietz R, Luft FC, Haller H. Amelioration of angiotensin II-induced cardiac injury by a 3-hydroxy-3-methylglutaryl coenzyme a reductase inhibitor. Circulation. 2001; 104: 576–581.
[Abstract/Free Full Text] - Hayashidani S, Tsutsui H, Shiomi T, Suematsu N, Kinugawa S, Ide T, Wen J, Takeshita A. Fluvastatin, a 3-hydroxy-3-methylglutaryl coenzyme a reductase inhibitor, attenuates left ventricular remodeling and failure after experimental myocardial infarction. Circulation. 2002; 105: 868–873.
[Abstract/Free Full Text] - Takemoto M, Node K, Nakagami H, Liao Y, Grimm M, Takemoto Y, Kitakaze M, Liao JK. Statins as antioxidant therapy for preventing cardiac myocyte hypertrophy. J Clin Invest. 2001; 108: 1429–1437.[CrossRef][Medline] [Order article via Infotrieve]
- Eberlein M, Heusinger-Ribeiro J, Goppelt-Struebe M. Rho-dependent inhibition of the induction of connective tissue growth factor (CTGF) by HMG CoA reductase inhibitors (statins). Br J Pharmacol. 2001; 133: 1172–1180.[CrossRef][Medline] [Order article via Infotrieve]
- Nogaki F, Muso E, Yashiro M, Kasuno K, Kamata T, Ono T, Sasayama S. Direct inhibitory effects of simvastatin on matrix accumulation in cultured murine mesangial cells. Kidney Int Suppl. 1999; 71: S198–S201.[Medline] [Order article via Infotrieve]
- Riessen R, Axel DI, Fenchel M, Herzog UU, Rossmann H, Karsch KR. Effect of HMG-CoA reductase inhibitors on extracellular matrix expression in human vascular smooth muscle cells. Basic Res Cardiol. 1999; 94: 322–332.[CrossRef][Medline] [Order article via Infotrieve]
- Ruiz-Ortega M, Rodriguez-Vita J, Sanchez-Lopez E, Carvajal G, Egido J. TGF-beta signaling in vascular fibrosis. Cardiovasc Res. 2007; 74: 196–206.
[Abstract/Free Full Text] - Mori T, Kawara S, Shinozaki M, Hayashi N, Kakinuma T, Igarashi A, Takigawa M, Nakanishi T, Takehara K. Role and interaction of connective tissue growth factor with transforming growth factor-beta in persistent fibrosis: A mouse fibrosis model. J Cell Physiol. 1999; 181: 153–159.[CrossRef][Medline] [Order article via Infotrieve]
- Turner NA, O’Regan DJ, Ball SG, Porter KE. Simvastatin inhibits MMP-9 secretion from human saphenous vein smooth muscle cells by inhibiting the RhoA/ROCK pathway and reducing MMP-9 mRNA levels. FASEB J. 2005; 19: 804–806.
[Abstract/Free Full Text] - Ohtsu H, Suzuki H, Nakashima H, Dhobale S, Frank GD, Motley ED, Eguchi S. Angiotensin II signal transduction through small GTP-binding proteins: mechanism and significance in vascular smooth muscle cells. Hypertension. 2006; 48: 534–540.
[Free Full Text] - Yamakawa T, Tanaka S, Numaguchi K, Yamakawa Y, Motley ED, Ichihara S, Inagami T. Involvement of Rho-kinase in angiotensin II-induced hypertrophy of rat vascular smooth muscle cells. Hypertension. 2000; 35: 313–318.
[Abstract/Free Full Text] - Takeda K, Ichiki T, Tokunou T, Iino N, Fujii S, Kitabatake A, Shimokawa H, Takeshita A. Critical role of Rho-kinase and MEK/ERK pathways for angiotensin II-induced plasminogen activator inhibitor type-1 gene expression. Arterioscler Thromb Vasc Biol. 2001; 21: 868–873.
[Abstract/Free Full Text] - Funakoshi Y, Ichiki T, Shimokawa H, Egashira K, Takeda K, Kaibuchi K, Takeya M, Yoshimura T, Takeshita A. Rho-kinase mediates angiotensin II-induced monocyte chemoattractant protein-1 expression in rat vascular smooth muscle cells. Hypertension. 2001; 38: 100–104.
[Abstract/Free Full Text] - Wang YX, Da CV, Martin-McNulty B, Vincelette J, Li W, Choy DF, Halks-Miller M, Mahmoudi M, Schroeder M, Johns A, Light DR, Dole WP. Inhibition of Rho-kinase by fasudil attenuated angiotensin II-induced cardiac hypertrophy in apolipoprotein E deficient mice. Eur J Pharmacol. 2005; 512: 215–222.[CrossRef][Medline] [Order article via Infotrieve]
- Mukai Y, Shimokawa H, Matoba T, Kandabashi T, Satoh S, Hiroki J, Kaibuchi K, Takeshita A. Involvement of Rho-kinase in hypertensive vascular disease: a novel therapeutic target in hypertension. FASEB J. 2001; 15: 1062–1064.
[Free Full Text] - Uehata M, Ishizaki T, Satoh H, Ono T, Kawahara T, Morishita T, Tamakawa H, Yamagami K, Inui J, Maekawa M, Narumiya S. Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature. 1997; 389: 990–994.[CrossRef][Medline] [Order article via Infotrieve]
- Kataoka C, Egashira K, Inoue S, Takemoto M, Ni W, Koyanagi M, Kitamoto S, Usui M, Kaibuchi K, Shimokawa H, Takeshita A. Important role of Rho-kinase in the pathogenesis of cardiovascular inflammation and remodeling induced by long-term blockade of nitric oxide synthesis in rats. Hypertension. 2002; 39: 245–250.
[Abstract/Free Full Text] - Ni W, Egashira K, Kataoka C, Kitamoto S, Koyanagi M, Inoue S, Takeshita A. Antiinflammatory and antiarteriosclerotic actions of HMG-CoA reductase inhibitors in a rat model of chronic inhibition of nitric oxide synthesis. Circ Res. 2001; 89: 415–421.
[Abstract/Free Full Text] - Eto Y, Shimokawa H, Hiroki J, Morishige K, Kandabashi T, Matsumoto Y, Amano M, Hoshijima M, Kaibuchi K, Takeshita A. Gene transfer of dominant negative Rho kinase suppresses neointimal formation after balloon injury in pigs. Am J Physiol Heart Circ Physiol. 2000; 278: H1744–H1750.
[Abstract/Free Full Text] - Seko T, Ito M, Kureishi Y, Okamoto R, Moriki N, Onishi K, Isaka N, Hartshorne DJ, Nakano T. Activation of RhoA and inhibition of myosin phosphatase as important components in hypertension in vascular smooth muscle. Circ Res. 2003; 92: 411–418.
[Abstract/Free Full Text] - Laufs U, Kilter H, Konkol C, Wassmann S, Bohm M, Nickenig G. Impact of HMG CoA reductase inhibition on small GTPases in the heart. Cardiovasc Res. 2002; 53: 911–920.
[Abstract/Free Full Text] - Banes-Berceli AK, Shaw S, Ma G, Brands M, Eaton DC, Stern DM, Fulton D, Caldwell RW, Marrero MB. Effect of simvastatin on high glu. Am J Physiol Renal Physiol. 2006; 291: F116–F121.
[Abstract/Free Full Text] - Ortego M, Bustos C, Hernandez-Presa MA, Tunon J, Diaz C, Hernandez G, Egido J. Atorvastatin reduces NF-kappaB activation and chemokine expression in vascular smooth muscle cells and mononuclear cells. Atherosclerosis. 1999; 147: 253–261.[CrossRef][Medline] [Order article via Infotrieve]
- Wassmann S, Laufs U, Muller K, Konkol C, Ahlbory K, Baumer AT, Linz W, Bohm M, Nickenig G. Cellular antioxidant effects of atorvastatin in vitro and in vivo. Arterioscler Thromb Vasc Biol. 2002; 22: 300–305.
[Abstract/Free Full Text] - Shirai H, Autieri M, Eguchi S. Small GTP-binding proteins and mitogen-activated protein kinases as promising therapeutic targets of vascular remodeling. Curr Opin Nephrol Hypertens. 2007; 16: 111–115.[Medline] [Order article via Infotrieve]
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||