Published online before print April 24, 2008, doi: 10.1161/ATVBAHA.108.165688
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© 2008 American Heart Association, Inc.
Integrative Physiology/Experimental Medicine |
Alpha2-Antiplasmin Is a Critical Regulator of Angiotensin II–Mediated Vascular Remodeling
From the Department of Physiology (Y.Z.H., K.O., C.O., O.M.), Kinki University School of Medicine, Osaka, Japan; and the Department of Food Science and Nutrition (S.U.), Kinki University School of Agriculture, Nara, Japan; and the Department of Microbiology and Infectious Diseases, University of Calgary, Calgary, Alberta T2N 4N1, Canada.
Osamu Matsuo and Yongzhong Hou, Department of Physiology, Kinki University School of Medicine, Osaka, Japan; and the Department of Food, Science and Nutrition (S.U.), Kinki University School of Agriculture, Nara, Japan. E-mail matsuo-o{at}med.kindai.ac.jp and hou_yz@yahoo.com
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Abstract |
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Objective— Alpha2-antiplasmin (
2-AP) is the major circulating inhibitor of plasmin, which plays a determining role in the regulation of intravascular fibrinolysis. We investigated the role of 2-AP on vascular remodeling in response to angiotensin II (Ang II).
Methods and Results— 2-AP–deficient mice were performed. Ang II and N
-nitro- L-arginine methyl ester (L-NAME)–induced perivascular fibrosis was significantly decreased in 2-AP–/– mice compared with wild-type mice. In situ gelatinolytic activity analysis shows that perivascular gelatinolytic activity was increased in 2-AP–/– mice, which was responsible for decreased perivascular fibrosis in response to Ang II and L-NAME. Ang II–induced arterial wall thickening, vascular cell proliferation, apoptosis, c-Myc, and collagen I expression were significantly decreased in 2-AP–/– mice compared with wild-type mice. Further analysis shows that increased p53 and p21 expression were responsible for inhibition of Ang II–induced vascular remodeling in 2-AP–/– mice.
Conclusion— The results show that 2-AP is a critical regulator for vascular remodeling by inhibiting p53/p21 pathway, suggesting that 2-AP is proposed to be a potential therapeutic target for vascular remodeling.
2-AP is the major circulating inhibitor of plasmin. Here we reported that 2-AP–/– suppressed Ang II and L-NAME–induced perivascular fibrosis by decreased collagen I expression. 2-AP–/– decreased Ang II–induced vascular remodeling by inhibiting p53/p21 pathway, suggesting that 2-AP is proposed to be a potential therapeutic target for vascular remodeling.
Key Words: Ang II • L-NAME • alpha2-antiplasmin • vascular remodeling • perivascular fibrosis
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Introduction |
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Angiotensin II (Ang II) is a vasoactive peptide with a wide variety of cardiovascular effects. Inhibition of Ang II by angiotensin-converting enzyme (ACE) inhibitors or angiotensin receptor antagonists prevents heart failure progression and reduces mortality in patients. As a critical hormone, Ang II plays a crucial role in affecting the function of virtually all organs, including heart, kidney, vasculature, and brain.1 Chronic Ang II stimulation promotes cardiac hypertrophy, vascular remodeling, and fibrosis.2,3
Alpha2-antiplasmin (2-AP) is the major circulating inhibitor of plasmin, which plays a determining role in the regulation of intravascular fibrinolysis. Alpha2-AP can be synthesized in a number of tissues, where it could function as a distal regulator of plasmin-mediated extracellular proteolysis.4,5 The 2-AP of human and murine is serpins (serine protease inhibitor) with molecular weight of 65 to 70 kDa.6 It forms a conventional serpin/enzyme complex with plasmin, but its activity is modified by the presence of N- and C-terminal extensions that are unique among the serpin family. The rate of association of 2-AP with plasmin is extremely fast. This is comparable to the rate of association of antithrombin and thrombin in the presence of unfractionated heparin.5 The fibrinolytic system plays a pivotal role in embryogenesis, ovulation, intima formation, proliferation, migration, tumorigenesis, and metastasis.7 The plasmin/2-AP complex in plasma is increased in acute stroke, myocardial infarction, unstable angina, and arterial fibrillation.8,9 The mutant 2-AP (P1 Arg-Ala), which has no plasmin inhibitory function, is added to human plasma or whole blood clots, leading to enhance urokinase (UK)-induced clot lysis.10 Similarly, a monoclonal antibody of 2-AP is added to plasma, leading to enhance the effectiveness of tPA-induced clot lysis.11 Lack of 2-AP inhibits neointima formation after vascular injure,12 and our previous investigation shows that lack of 2-AP attenuates dermal fibrosis.13 However, 2-AP–induced vascular remodeling in response to Ang II is still unknown. We reported here that Ang II–mediated vascular remodeling was decreased in 2-AP–deficient mice. These findings suggest that 2-AP is a key regulator in Ang II–infused mice.
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Materials and Methods |
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Animals
2-AP–deficient mice were generated by injection of 2-AP–deficient 129/Sv ES cells into C57BL/6 blastocysts to generate chimeras, and heterozygous males were bred with C57BL/6 females as described previously.14 All experiments were performed using male 2-AP–/– mice with littermate 2-AP+/+ controls. At least 6 pairs were prepared for every experiment. Animal care and work protocols were approved and carried out following the regulations by the Committee of Animal Research and Bioethics of Kinki University. Unless otherwise stated, we used 8- to 10-week-old C57BL/6 mice in the experiments. All experiments were performed in accordance with institutional and national guidelines.
Drug Treatment and Blood Pressure Analysis
Ang II (Sigma-Aldrich) was dissolved in 0.9% saline. Mice were treated with either Ang II (1.4 mg/kg/d) or vehicle (0.9% NaCl) through subcutaneous osmotic minipumps (Durect) for 2 weeks. Mice were administered L-NAME (Sigma-Aldrich), an NOS inhibitor, in the drinking water (120 mg/kg/d) for 8 weeks as described previously,15 whereas control animals received unmodified drinking water. All animals were fed a regular chow diet. Systolic and diastolic blood pressure and heart rate were measured in conscious mice by the tail-cuff system using BP98A (Softron Co) according to manufacturer’s protocol.
Reverse Transcription PCR Analysis
The mRNA of mice 2-AP was analyzed by reverse transcription PCR. The thoracic arterial tissues for RNA isolation (20 to 30 µg) were homogenized, and total RNA was isolated by using RNeasy Mini kit (Qiagen) according to the manufacturer’s specifications. Total RNA (0.5 µg) from each sample was reverse transcribed. Reverse transcription PCR was performed by using Excript TR reagent kit (TaKaRa). Amplified PCR products were resolved in 2% agarose gels, stained with ethidium bromide, and photographed under UV light. For PCR amplification, the following primers were used: GAPDH forward, 5'-TGCATCCTGCACCACCAACT; GAPDH reverse, 5'-AACACGGAAGG CCATGCCAG-3'; 2-AP primer was described previously.16
Histological Analysis
Thoracic arterial or heart tissues were paraffin embedded and cut into 5-µm sections. Sections were stained with hematoxylin and eosin (HE). Picrosirius red staining was used to detect perivascular fibrosis. Cell apoptosis was detected by using terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) analysis kit (Wako).
Immunohistochemistry
Sections (5 µm) cut from paraffin-embedded arterial sections were stained with antibodies against p21, p53, c-Myc, Bcl2 (Santa Cruz). We stained all sections with biotinylated secondary antibodies and detected by using Vectastain Elite ABC kits (Vector Laboratories).
PCNA and BrdU Staining
Proliferating cell nuclear antigen (PCNA; Cell signaling) staining was performed on 5-µm paraffin-embedded arterial sections. BrdU (100 mg/kg in saline, sigma) was injected intraperitoneally at 1, 8, 16, 24 hours before removal of arterial tissues as described previously,12 and then thoracic arterial tissues were fixed in 10% neutral buffered formalin. Immunohistochemical staining was performed by using BrdU antibody (Santa Cruz). The labeling index was calculated as ratio of the number of positive nuclei to the number of total nuclei stained.
In Situ Gelatinolytic Activity Assay
In situ gelatinolytic activity was assessed as described previously.17 After 2 weeks of Ang II infusion or eight weeks of L-NAME treatment, animals were euthanized, and thoracic arterial tissues were frozen. Arterial sections, 10 µm thick, were incubated for 2 hours at 37°C in a solution of gelatin fluorescein conjugate (40 µg/mL) (Molecular Probes). This procedure was performed carefully without any washing. In situ gelatinolytic activity was assesed by fluoromicroscopic detection.
Western Blot
Thoracic arterial tissues were minced and homogenated in lysis buffer (50 mmol/L HEPES, pH 7.0, 150 mmol/L NaCl, 10% Glycerol, 1% Triton X-100, 1.5 mmol/L MgCl2, 1 mmol/L EGTA, 100 mmol/L NaF, 10 mmol/L NaPPi, 1 mmol/L Na3VO4, 1 mmol/L PMSF, 10 µg/mL aprotinin and 10 µg/mL leupeptin). The samples were subjected to 10% to 20% gradient SDS-PAGE, transferred to a nitrocellulose membrane, then probed by Western blot analysis with the indicated antibodies (Santa Cruz) and developed by using ECL Kit (Amersham Biosciences).
Statistical Analysis
The values for each parameter within a group are expressed as the mean±SEM. Statistical comparison was carried out with 1-way analysis of variance (ANOVA) and Dunnett test. Significance was defined as P<0.05.
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Results |
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Involvement of
2-AP in Ang II–Induced Vascular RemodelingAlpha2-AP can be synthesized in a number of tissues,4 however it is still unknown whether Ang II increases
2-AP expression in arterial tissues. Our results show that 2-AP gene expression was very low in thoracic arterial tissues, but its expression was significantly increased after Ang II infusion for 2 weeks (Figure 1A). Chronic infusion of Ang II in animals leads to increased systolic blood pressure and vascular remodeling, including medial hypertrophy and perivascular fibrosis, which are also observed in patients with renovascular hypertension.1 There was no difference in the systolic blood pressure observed in 2-AP–/– mice compared with 2-AP+/+ mice in response to Ang II (Figure 1B). Ang II infusion led to increase vessel thickening and the ratio of the media to vessel lumen, but significant blunting of these Ang II–mediated effects was observed in 2-AP–/– mice (Figure 1C through 1E). These results show that vascular remodeling was decreased in Ang II–induced 2-AP–/– mice.
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Loss of 2-AP Decreases Perivascular Fibrosis in Response to Ang II or L-NAME
Compared with saline-infused wild-type (WT) mice, there was a significant increase in perivascular fibrosis as measured by Picrosirius red staining of arterial and ventricular sections from Ang II–infused WT mice (Figure 2A and 2D). In contrast, perivascular fibrosis was decreased in Ang II–infused 2-AP–/– mice (Figure 2A and 2D). Long-term inhibition of nitric oxide synthase (NOS) by use of L-NAME induces perivascular fibrosis in experimental animal models, and perivascualr fibrosis is decreased in plasminogen activator inhibitor (PAI)-1–/– mice.15 We further detected perivascular fibrosis and systolic blood pressure in response to L-NAME. Our results show that although perivasular fibrosis and systolic blood pressure were increased in L-NAME–treated mice, significant blunting of these L-NAME–mediated effects was observed in 2-AP–/– mice (Figure 3A and 3C). Further analysis shows that collagen I expression of 2-AP–/– mice was decreased in response to Ang II or L-NAME (Figure 2E and Figure 3D).
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Loss of 2-AP Increases Perivascular Gelatinolytic Activity
Matrix metalloproteinases (MMPs) are associated with embryogenesis, wound healing, inflammation, fibrosis, and cardiovascular diseases.18 To further detect the effect of 2-AP–/– mice on MMPs in response to Ang II or L-NAME, in situ gelatinolytic activity was analyzed. Our results show that perivascular gelatinolytic activity was increased in 2-AP–/– mice, which was responsible for inhibition of Ang II or L-NAME–induced perivascular fibrosis (Figure 4A and 4B).
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Involvement of 2-AP in Ang II–Induced Vascular Cell Proliferation
To detect and quantify vascular cell proliferation, the staining of proliferation cell nuclear antigen (PCNA) and BrdU were performed by immunostaining analysis. There were only a few positive nuclei for PCNA and BrdU staining in vascular wall in 2-AP+/+ mice and 2-AP–/– mice with saline treatment (Figure 5A and 5B). Although there was a marked increase in the number of PCNA- and BrdU-positive cells in vascular wall in Ang II–treated 2-AP+/+ mice, the number of PCNA- and BrdU-stained positive cells of 2-AP–/– mice was significantly decreased in response to Ang II (Figure 5A and 5B). The results suggest that vascular hyperplasia resulted from increased proliferation of vascular mesenchymal cells.
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Effects of p21 and c-Myc Expression in Ang II–Induced WT and 2-AP–Deficient Mice
The number of PCNA- and BrdU-stained cells of 2-AP–/– mice was significantly decreased in response to Ang II, suggesting that loss of 2-AP resulted in inhibiting Ang II–induced vascular cell proliferation. As the cell cycle inhibitor, p21 inhibits vascular smooth muscle cell (VSMC) proliferation.19 Immunohistochemical analysis shows that Ang II significantly decreased p21 protein expression, but this effect was decreased in Ang II–induced 2-AP–/– mice (supplemental Figure IA, available online at http://atvb.ahajournals.org). Our previous investigation shows that c-Myc induces VSMC proliferation.20 Here we found that Ang II significantly increased c-Myc expression. In contrast, c-Myc expression of 2-AP–/– mice was decreased in response to Ang II (supplemental Figure IB).
Loss of 2-AP Inhibits Vascular Cell Apoptosis by Increased Bcl2 Expression
Although Ang II increased p53 expression both in WT mice and 2-AP–/– mice (supplemental Figure IIA), Bcl2 expression of 2-AP–/– mice was significantly increased in response to Ang II (supplemental Figure IIB), which was responsible for inhibition of Ang II–induced cell apoptosis (supplemental Figure IIC and IID).
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Discussion |
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The purpose of this study was to determine whether loss of
2-AP decreases Ang II–induced vascular remodeling. Our findings suggest that 2-AP plays a crucial role in development of vascular remodeling in response to Ang II. Arterial wall thickening and perivascular fibrosis were significantly decreased in 2-AP–/– mice compared with WT mice in response to Ang II.
Although Ang II is a potent vasoactive peptide, several of its effects occur independently of changes in blood pressure. Our results show that Ang II increased blood pressure in both 2-AP+/+ mice and 2-AP–/– mice in response to Ang II; these findings further suggest that many of the cellular effects mediated by Ang II occur independently of the vasoactive effects.3,21,22,23
Ang II promotes fibroblast proliferation, alteration of fibrillar collagen turnover, and stimulation of aldosterone, resulting in accumulation of collagen type I and III fibers and fibrosis24; this accumulation causes a distortion of tissue structure, which is responsible for the increase in myocardial stiffness leading to diastolic dysfunction, a substrate for ventricular arrhythmias, and ultimately to systolic dysfunction.25 Matrix metalloproteinases (MMPs), also called matrixins, are a family of zinc-dependent endopeptidases capable of degrading extracellular matrix components such as collagens, proteoglycans, elastin, laminin, fibronectin, and other glycoproteins.26 The plasminnogen activator/plasmin system plays a crucial role in degrading extracellular matrix components, because plasmin is the proteolytic amplification that can be achieved by activating several MMPs.27 The serine proteinase inhibitor, 2-AP, is a member of the serpin family with plasmin as its primary target. Plasmin, generated from the zymogen plasminogen, plays a critical role in fibrin proteolysis and tissue remodeling.28 These findings suggest that 2-AP deficiency may prevent the increase of collagen deposition by accelerating matrix degradation. Thus, 2-AP could be an important modulator during the process of vascular remodeling by regulating the activation of MMPs. On the other hand, long-term inhibition of NOS by use of L-NAME induces perivascular fibrosis in experimental animal models, and PAI-1–/– inhibits L-NAME–induced coronary arterial fibrosis by increased gelatinolytic activity.29 Here we found that loss of 2-AP decreased perivascular fibrosis in response to Ang II or L-NAME. In situ gelatinolytic activity analysis suggests that perivascular gelatinolytic activity of 2-AP–/– mice was significantly increased. Further analysis shows that Ang II or L-NAME–induced collagen I expression was decreased in 2-AP–/– mice. These findings suggest that loss of 2-AP decreased Ang II or L-NAME–induced perivascular fibrosis. Our results show that 2-AP plays a crucial role in cardiovascular diseases. Gelatinolytic activity has the ability to cleave a variety of the extracellular matrix (ECM) components including denatured collagen (gelatin) generated by thermal denaturation at body temperature.30 Therefore, increased perivascular gelatinolytic activity was responsible for inhibition of fibrosis development in response to Ang II or L-NAME.
Antimitogenic signals activate p53, which induces expression of CKI p21CIP1 and consequently inhibits the activity of the G1 cyclin-CDK complexes, resulting in G1-phase arrest.31 The p21 gene, another member of the CIP/KIP family of CDK inhibitors, also is an important modulator in the regulation of cell cycle progression.32 Overexpression of p21 has been associated with a reduction in systemic arterial smooth muscle cell proliferation.33 Expression of p21 inhibits vascular cell proliferation and induces cell cycle arrest.34 In addition, inhibition of heparin on pulmonary vascular pericyte proliferation is accompanied by induction of p21.35 Our results show that Ang II decreased p21 expression in WT mice. In contrast, this effect was abolished in 2-AP–/– mice. Ang II induces VSMC proliferation and apoptosis, which is responsible for vascular remodeling and atherosclerosis.1 Although Ang II induced vascular cell apoptosis in WT mice, the apoptosis of 2-AP–/– mice was inhibited. Further analysis shows that loss of 2-AP led to increase Bcl2 expression in response to Ang II. Bcl2 is antiapoptotic protein, and inhibition of apoptosis is associated with increased Bcl2 expression in response to Ang II.36 These findings show that Bcl2 expression in Ang II–treated 2-AP–/– mice was responsible for inhibition of cell apoptosis. These results provide a novel role for 2-AP–mediated vascular remodeling by suppressing p52/p21 pathway in response to Ang II.
In conclusion, our results show that loss of 2-AP alleviated Ang II–induced vascular remodeling in association with increased p53/p21 pathway. Increased perivascular gelatinolytic activity and decreased collagen I expression in 2-AP–/– mice contributed to inhibit the development of perivascular fibrosis. These findings suggest that 2-AP is proposed to be a potential therapeutic target for vascular remodeling.
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Acknowledgments |
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Sources of Funding
This work was supported by following funds from the Ministry of Education, Culture, Sports, Science and Technology: Grant-in-Aid for Scientific Research No.19590217, "High-Tech Research Center Project for Private Universities", and Medical and Engineering Link Project.
Disclosures
None.
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Footnotes |
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Original received December 9, 2007; final version accepted April 9, 2008.
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References |
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1. Mehta PK, Griendling KK. Angiotensin II cell signaling: physiological and pathological effects in the cardiovascular system. Am J Physiol Cell Physiol. 2007; 292: 82–97.[CrossRef]
2. Zhan Y, Brown C, Maynard E, Anshelevich A, Ni W, Ho IC, Oettgen P. Ets-1 is a critical regulator of Ang II–mediated vascular inflammation and remodeling. J Clin Invest. 2005; 115: 2508–2516.[CrossRef][Medline] [Order article via Infotrieve]
3. Izumiya Y, Kim S, Izumi Y, Yoshida K, Yoshiyama M, Matsuzawa A, Ichijo H, Iwao H. Apoptosis signal-regulating kinase 1 plays a pivotal role in angiotensin II induced cardiac hypertrophy and remodeling. Circ Res. 2003; 93: 874–883.
4. Menoud PA, Sappino N, Boudal-Khoshbeen M, Vassalli JD, Sappino AP. The kidney is a major site of alpha(2)-antiplasmin production. J Clin Invest. 1996; 97: 2478–2484.[Medline] [Order article via Infotrieve]
5. Coughlin PB. Antiplasmin: The forgotten serpin? FEBS J. 2005; 272: 4852–4857.[CrossRef][Medline] [Order article via Infotrieve]
6. Wiman B, Collen D. Purification and characterization of human antiplasmin, the fast-acting plasmin inhibitor in plasma. Eur J Biochem. 1977; 78: 19–26.[Medline] [Order article via Infotrieve]
7. Carmeliet P, Collen D. Gene manipulation and transfer of the plasminogen and coagulation system in mice. Semin Thromb Hemost. 1996; 22: 525–542.[Medline] [Order article via Infotrieve]
8. Montes R, Paramo JA, Angles-Cano E, Rocha E. Development and clinical application of a new ELISA assay to determine plasmin-alpha2-antiplasmin complexes in plasma. Br J Haematol. 1996; 92: 979–985.[CrossRef][Medline] [Order article via Infotrieve]
9. Bayes-Genis A, Guindo J, Oliver A. Elevated levels of plasmin-alpha 2 antiplasmin complexes in unstable angina. Throm Haemost. 1999; 81: 865–868.[Medline] [Order article via Infotrieve]
10. Lee KN, Tae WC, Jackson KW, Kwon SH, McKee PA. Characterization of wild-type and mutant a2-antiplasmin: fibrinolysis enhancement by reactive site mutant. Blood. 1999; 94: 164–171.
11. Sakata Y, Eguchi Y, Mimuro J, Matsuda M, Sumi Y. Clot lysis induced by a monoclonal antibody against alpha2-plasmin inhibitor. Blood. 1989; 74: 2692–2697.
12. Matsuno H, Ishisaki A, Nakajima K, Okada K, Ueshima S, Matsuo O, Kozawa O. Lack of {alpha}2-antiplasmin promotes re-endothelialization via over-release of VEGF after vascular injury in mice. Blood. 2003; 102: 3621–3628.
13. Kanno Y, Kuroki A, Okada K, Tomogane K, Ueshima S, Matsuo O, Matsuno H. Alpha2-antiplasmin is involved in the production of transforming growth factor beta1 and fibrosis. J Thromb Haemost. 2007; 5: 2266–2273.[CrossRef][Medline] [Order article via Infotrieve]
14. Okada K, Lijnen HR, Dewerchin M, Belayew A, Matsuo O, Collen D, Bernaerts R. Characterization and targeting of the murine 2-antiplasmin gene. Throm Haemost. 1997; 78: 1104–1110.[Medline]
[Order article via Infotrieve]
15. Kaikita K, Fogo AB, Ma L, Schoenhard JA, Brown NJ, Vaughan DE. Plasminogen activator inhibitor-1 deficiency prevents hypertension and vascular fibrosis in response to long-term nitric oxide synthase inhibition. Circulation. 2001; 104: 839–844.
16. Lijnen HR, Okada K, Matsuo O, Collen D. Alpha2-antiplasmin gene deficiency in mice is associated with enhanced fibrinolytic potential without overt bleeding. Blood. 1999; 93: 2274–2281.
17. Gasche Y, Copin JC, Sugawara T, Fujimura M, Chan PH. Matrix metalloproteinase inhibition prevents oxidative stress-associated blood-brain barrier disruption after transient focal cerebral ischemia. J Cereb Blood Flow Metab. 2001; 21: 1393–1400.[CrossRef][Medline] [Order article via Infotrieve]
18. Chakraborti S, Mandal M, Das S, Mandal A, Chakraborti T. Regulation of matrix metalloproteinases: an overview. Mol Cell Biochem. 2003; 253: 269–285.[CrossRef][Medline] [Order article via Infotrieve]
19. Quasnichka H, Slater SC, Beeching CA, Boehm M, Sala-Newby GB, George SJ. Regulation of smooth muscle cell proliferation by beta-catenin/T-cell factor signaling involves modulation of cyclin D1 and p21 expression. Circ Res. 2006; 99: 1329–1337.
20. Hou YZ, Okamoto C, Okada K, K, Naoyaki K, Ueshima S, Matsu O. c-Myc is essential for urokinase plasminogen activator expression on hypoxia induced vascular smooth muscle cells. Cardiovas Res. 2007; 75: 186–194.
21. Satoh M, Ogita H, Takeshita K, Mukai Y, Kwiatkowski DJ, Liao JK. Requirement of Rac1 in the development of cardiac hypertrophy. Proc Natl Acad Sci USA. 2006; 103: 7432–7437.
22. Huggins CE, Domenighetti AA, Pedrazzini T, Pepe S, Delbridge LM. Elevated intracardiac angiotensin II leads to cardiac hypertrophy and mechanical dysfunction in normotensive mice. J Renin Angiotensin Aldosterone Syst. 2003; 4: 186–190.
23. Rugale C, Delbosc S, Cristol JP, Mimran A, Jover B. Sodium restriction prevents cardiac hypertrophy and oxidative stress in angiotensin II hypertension. Am J Physiol Heart Circ Physiol. 2003; 284: H1744–H1750.
24. Campbell JE, Janicki JS, Weber KT. Temporal differences in fibroblast proliferation and phenotype expression in response to chronic administration of angiotensin II or aldosterone. J Mol Cell Cardiol. 1995; 27: 1545–1560.[CrossRef][Medline] [Order article via Infotrieve]
25. Weber KT, Brilla CG, Janicki S. Myocardial fibrosis: functional signifi cance and regulatory factors. Cardiovasc Res. 1993; 27: 341–348.
26. Sluijter JP, de Kleijn DP, Pasterkamp G. Vascular remodeling and protease inhibition-bench to bedside. Cardiovas Res. 2006; 69: 595–603.
27. Creemers EEJM, Cleutjens J, Smits JFM, Daemen MJAP. Matrix metalloproteinases after myocardial infarctiona new approach to prevent heart failure. Circ Res. 2001; 89: 201–210.
28. Collen D. The plasminogen (fibrinolytic) system. Thromb Haem. 1999; 82: 259–270.[Medline] [Order article via Infotrieve]
29. Kaikita K, Schoenhard JA, Painter CA, Ripley RT, Brown NJ, Fogo AB, Vaughan DE. Potential roles of plasminogen activator system in coronary vascular remodeling induced by long-term nitric oxide synthase inhibition. J Mol Cell Cardiol. 2002; 34: 617–627.[CrossRef][Medline] [Order article via Infotrieve]
30. Shipley JM, Doyle GA, Fliszar CJ, Ye QZ, Johnson LL, Shapiro SD, Welgus HG, Senior RM. The structural basis for the elastolytic activity of the 92-kDa and 72-kDa gelatinases. Role of the fibronectin type II-like repeats. J Biol Chem. 1996; 271: 4335–4341.
31. Levine AJ. p53, the cellular gatekeeper for growth and division. Cell. 1997; 88: 323–331.[CrossRef][Medline] [Order article via Infotrieve]
32. Nathe TJ, Deou J, Walsh B, Bourns B, Clowes AW, Daum G. Interleukin-1 beta inhibits expression of p21(WAF1/CIP1) and p27(KIP1) and enhanced proliferation in response to platelet-derived growth factor-BB in smooth muscle cells. Arterioscler Throm Vasc Biol. 2002; 22: 1293–1298.
33. Tanner FC, Boehm M, Akyurek LM, San H, Yang ZY, Tashiro J, Nabel GJ, Nabel EG. Deferential effeots of the cyclindependent kinase inhibitors p27kip, p21cip1, p16ink4 on vascular smooth muscle cell proliferation. Circulation. 2000; 101: 2022–2025.
34. Yang ZY, Simari RD, Perkins ND, San H, Gordon D, Nabel GJ, Nabel EG. Role of the p21 cyclin-dependent kinase inhibitor in limiting intimal cell proliferation in response to arterial injury. Proc Natl Acad Sci USA. 1996; 9: 7905–7910.
35. Khoury J, Langleben D. Heparin-like molecules inhibit pulmonary vascular periocyte proliferation in vitro. Am J Physiol Lung Cell Mol Physiol. 2000; 279: L252–L261.
36. Grishko V, Pastukh V, Solodushko V, Gillespie M, Azuma J, Schaffer S. Apoptotic cascade initiated by angiotensin II in neonatal cardiomyocytes: role of DNA damage. Am J Physiol Heart Circ Physiol. 2003; 285: H2364–2372.
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