© 2000 American Heart Association, Inc.
Vascular Biology |
Enhanced Expression of Osteopontin in Human Diabetic Artery and Analysis of Its Functional Role in Accelerated Atherogenesis
From the Second Department of Internal Medicine (M.T., K.Y., T.M., Y.S., S.M.), School of Medicine, Chiba University, and the Department of Internal Medicine (M.N., T.S., T.H.), Sakura National Hospital, Chiba, Japan; and the Section of Immunopathogenesis (S.K., T.U.), Institute of Immunological Science, Hokkaido University, Hokkaido, Japan.
Correspondence to Minoru Takemoto, Second Department of Internal Medicine, School of Medicine, Chiba University, 1-8-1 Inohana, Chiba 260-0856, Japan. E-mail mtakemo{at}intmed02.m.chiba-u.ac.jp
 |
Abstract |
|---|
 Top Abstract IntroductionMethodsResultsDiscussionReferences |
|---|
Abstract—We have previously reported that high glucose stimulates osteopontin (OPN) expression through protein kinase C–dependent pathways as well as hexosamine pathways in cultured rat aortic smooth muscle cells. The finding prompted us to study in vivo expression of OPN in diabetes mellitus. In the present study, we found by immunohistochemistry that medial layers of the carotid arteries of streptozotocin-induced diabetic rats and the forearm arteries of diabetic patients stained positively for OPN antibodies, whereas the staining from arteries of control rats and nondiabetic patients was negative. We also found that OPN stimulated the migration and enhanced platelet-derived growth factor (PDGF)-mediated DNA synthesis of cultured rat aortic smooth muscle cells. OPN and PDGF synergistically activated focal adhesion kinase as well as extracellular signal–regulated kinase; this finding seems to explain the OPN-induced enhancement of PDGF-mediated DNA synthesis. Taken together, our present results raise a possibility that OPN plays a role in the development of diabetic vascular complications.
Key Words: diabetic macroangiopathy • osteopontin • platelet-derived growth factor • vascular smooth muscle cells
|
Introduction |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
It is generally accepted that diabetic patients often suffer from atherosclerotic vascular diseases, such as ischemic heart disease and arteriosclerosis obliterans.1 2 It is also known that diabetic vascular lesions tend to undergo restenosis after angioplasty and that diffuse calcification of the affected arteries is a characteristic feature of diabetic vascular diseases.3 4 5 However, the reason for the accelerated atherogenesis in diabetes mellitus has not been fully elucidated.
Osteopontin (OPN) is a multifunctional phosphoprotein secreted by many cell types, such as osteoclasts, lymphocytes, macrophages, epithelial cells, and vascular smooth muscle cells (SMCs).6 Overexpression of OPN has been found in several physiological as well as pathological conditions, including immunologic disorders,7 neoplastic transformation,8 progression of metastases,8 formation of urinary stones,9 and wound healing.10
It has been reported that OPN protein and mRNA are expressed in the neointima as well as in calcified atheromatous plaque.11 A neutralizing antibody against OPN has been found to inhibit rat carotid neointimal formation after endothelial denudation.12 It has also been reported that OPN inhibits the calcification of vascular SMCs in culture.13 These reports have suggested that OPN contributes not only to the tissue calcification process but also to the development of atherosclerosis, especially in the process of intimal thickening.
We have recently reported that high glucose levels stimulate OPN expression through protein kinase C–dependent pathways as well as hexosamine pathways in cultured rat aortic SMCs.14 The present study was undertaken to gain more insight into the mechanism of diabetic vascular complications. We first demonstrate that OPN protein is highly expressed in the medial layers of the arteries of diabetic rats and patients. Furthermore, OPN stimulates migration and enhances platelet-derived growth factor (PDGF)-mediated DNA synthesis of SMCs, possibly by promoting the activation of focal adhesion kinase (FAK) as well as extracellular signal–regulated kinase (ERK). The present data suggest that OPN plays a role in the accelerated atherogenesis in diabetes mellitus.
|
Methods |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
Reagents
Recombinant OPN was affinity-purified by using an antibody column filled with a monoclonal anti-human OPN antibody (10A16, IBL) from conditioned medium of full-length human OPN cDNA-transfected CHO-K1 cells.15 Recombinant human PDGF-BB was purchased from R&D Systems. A monoclonal anti-rat OPN antibody (MPIIIB10) was from American Research Products. A monoclonal anti–
-smooth
muscle actin antibody (1A4) was from Sigma Chemical Co.
[Methyl-3H]thymidine was from Amersham
Pharmacia Biotech.
Tissue Preparation
Male Wistar rats weighing 
200 g were administered either 50
mg/kg streptozotocin (STZ, Sigma) or normal saline into tail veins as
described previously.16 Four weeks later, the carotid
arteries were thoroughly perfused with 4%
paraformaldehyde in PBS, excised, fixed with 4%
paraformaldehyde in PBS for 4 hours at 4°C, rinsed
with PBS, and incubated with 30% sucrose in PBS for 3 hours at 4°C.
Then the fixed arteries were mounted in Tissue-Tek embedding OCT
compound (Miles Inc), snap-frozen, and subsequently stored at -70°C
before sectioning.17 A piece of vascular wall was excised
from the forearm arteries of 6 patients with end-stage renal disease
during a shunt operation after obtaining written informed consent for
their excised arteries to be used in the present experiment. Of the
6 patients, 5 were men; the ages of the 6 patients ranged between 40
and 80 years, with a mean age of 62 years. Three of the patients were
diabetics, 2 patients had been treated by diet restriction only, and
the other patient had been treated by diet restriction and subcutaneous
insulin infusion. The samples were fixed in 4%
paraformaldehyde at 4°C for 24 hours, mounted in
Tissue-Tek embedding OCT compound, snap-frozen, and subsequently stored
at -70°C before sectioning.17
Immunohistochemistry
Immunohistochemistry was performed essentially as
described.18 For the identification of OPN, we used the
anti-human OPN antibody (10A16, 5 µg/mL) as well as the anti-rat OPN
antibody (MPIIIB10, 1:250 dilution), which also cross-reacts with human
OPN, according to the manufacturer’s instructions. For the
identification of SMCs, we used the anti–-smooth muscle actin
antibody (1A4, 1:1000 dilution). Calcium deposition was assessed by von
Kossa staining.19
Cell Culture
Rat aortic SMCs were isolated by the explant method from adult
male Wistar rats, and the primary culture and subculture were carried
out as described.20 Cells at the 4th to 9th passages were
used for the present study.
Migration Assay
A chemotactic response was assayed essentially as
described21 with use of a 96-well microchemotaxis chamber
(Neuro Probe). Cells that had migrated to the lower surface of the
membrane were fixed and stained, and the number of the cells was
estimated by using a microplate reader (TOSHO) at 600 nm. Experiments
were performed in quadruplicate and were repeated at least 3 times.
[3H]Thymidine Incorporation Assay
DNA synthesis was estimated by measuring
[3H]thymidine incorporation into
trichloroacetic acid–precipitable materials of cultured cells as
described.22 Experiments were performed in quadruplicate
and were repeated at least 3 times.
Assays for FAK and ERK
Confluent SMCs in 6-cm dishes were serum-starved for 18 hours.
Then the cells were treated with different concentrations of PDGF-BB
and OPN at 37°C for 10 minutes. After incubation, activities of
FAK23 and ERK24 were assayed in the cells
exactly as described. Densitometric analyses of the detected
bands on gels were performed by use of an imaging scanner (EPSON ES
8000) with NIH Image 1.44 software.
Statistical Analysis
Data are expressed as mean±SEM. The significance of differences
was evaluated by Student t test.
|
Results |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
OPN Protein Is Highly Expressed in Medial Layers of Carotid Arteries in STZ-Induced Diabetic Rats and Forearm Arteries in Diabetic Patients
We have previously found that high glucose levels potently enhance OPN expression in cultured rat aortic SMCs.14 The finding prompted us to study the in vivo expression of OPN in diabetes mellitus. We examined OPN expression in STZ-induced diabetic rats by immunohistochemistry with use of a monoclonal anti-rat OPN antibody (MPIIIB10). Two diabetic rats and 2 control rats were examined. The mean blood glucose level of the diabetic rats (29.6 mmol/L) was much higher than that of the control rats (9.3 mmol/L). As shown in Figure 1
, the staining of the medial
layers of the carotid arteries derived from both of the diabetic rats
was positive for the OPN antibody (panel B), whereas the staining of
the arteries from the control rats was negative (panel A). These
results led us to further examine OPN expression in human samples. A
piece of the forearm arteries was collected during shunt operations
from age-matched diabetic (n=3) and nondiabetic (n=3) patients. As
shown in Figure 1, medial layers of the samples derived from all
the diabetic patients stained positively for a monoclonal anti-human
OPN antibody (10A16, panel D), whereas the staining of samples from the
nondiabetic patients was negative (panel C). The results were confirmed
by using another monoclonal anti-OPN antibody (MPIIIB10). The
OPN-stained areas also stained positively with an anti–-smooth
muscle actin antibody (1A4; data not shown), suggesting that the OPN
was produced by SMCs. No obvious relation was found between OPN
expression and calcium deposition (data not shown).
|
OPN Not Only Stimulates Migration but Also Enhances
PDGF-BB–Mediated DNA Synthesis of SMCs
To evaluate a possible role of OPN in atherogenesis, biological
effects of recombinant human OPN on cultured rat aortic SMCs were
analyzed. We first examined the effect of OPN on the migration
of SMCs by modified Boyden chamber assay. As shown in Figure 2, OPN dose-dependently enhanced SMC
migration. The migratory response was confirmed to be chemotactic, as
assessed by checkerboard analyses (data not shown).
|
We next examined the effect of OPN on DNA synthesis of SMCs by a
[3H]thymidine incorporation assay. It was found
that OPN by itself did not stimulate DNA synthesis of SMCs (data not
shown). However, as shown in Figure 3, OPN enhanced DNA synthesis of SMCs, which were
simultaneously stimulated with low-dose (1 ng/mL)
PDGF-BB.
|
OPN-Induced Enhancement of PDGF-BB–Stimulated DNA Synthesis Is
Mediated Possibly by Promoting Activation of FAK and ERK
OPN is known to be a ligand for some integrins.25
Thus, we examined the effect of OPN on the activation of FAK, a
downstream mediator of the integrin signaling pathway. After treatment
of SMCs with PDGF-BB and/or OPN, the cell lysates were
immunoprecipitated with the anti-FAK antibody, and the samples were
subjected to Western blotting. The blot was first probed with an
anti-phosphotyrosine antibody. As shown in Figure 4, treatment with 1 ng/mL PDGF-BB did not
appreciably enhance phosphorylation of FAK (top panel,
lane 2). Likewise, treatment of OPN alone did not appreciably enhance
FAK phosphorylation (data not shown). However, in the
presence of 1 ng/mL PDGF-BB, OPN potently enhanced FAK
phosphorylation (top panel, lanes 3 and 4). The same
blot was then stripped and reprobed with the anti-FAK antibody. The
intensity of the 125-kDa FAK band was not different among the lanes
(middle panel).
|
The results led us to further examine the effect of OPN on activation
of ERK, a putative downstream mediator of FAK. SMCs were serum-starved
and stimulated with PDGF-BB and/or OPN. After stimulation, ERK was
collected by immunoprecipitation and subjected to in vitro kinase assay
in the presence of Elk-1. ERK activity was estimated by the degree of
Elk-1 phosphorylation, which was detected by
immunoblotting with the specific antibody against
phosphorylated Elk-1. As shown in Figure 5, treatment with 1 ng/mL PDGF-BB
activated ERK to some degree (lane 3). Treatment with OPN alone
also slightly activated ERK (lane 4), but in the presence of 1
ng/mL PDGF-BB, OPN potently activated ERK (lanes 5 and 6).
|
|
Discussion |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
In the present study, we show that OPN is highly expressed in medial layers of the carotid arteries in STZ-induced diabetic rats as well as in the forearm arteries of patients with diabetes mellitus (Figure 1
). Our present study also shows that OPN stimulates
migration and enhances PDGF-BB–mediated DNA synthesis of cultured rat
aortic SMCs (Figures 2 and 3). These results, together with our
previous finding that high glucose levels stimulate OPN expression in
cultured rat aortic SMCs,14 imply that OPN plays a role in
the development of diabetic vascular complications. In vivo expression of OPN has previously been analyzed in several diabetic animal models. Towler et al26 demonstrated the upregulation of OPN expression in the aortas of high-fat diet–induced diabetic mice. Fischer et al27 reported that the upregulation of OPN expression in the renal cortex of STZ-induced diabetic rats was mediated by bradykinin. We also found the enhanced expression of OPN mRNA in the aortas of STZ-induced diabetic rats (data not shown). However, data concerning OPN expression in human subjects with diabetes mellitus have not yet been reported. In the present study, we have demonstrated for the first time that OPN expression is also enhanced in human diabetic arteries, confirming that the diabetes-induced upregulation of OPN expression is a general phenomenon observed across species.
OPN was shown to promote chemotaxis of different cell types, including
vascular SMCs,28 macrophages,29 and
osteoclasts.30 OPN was also shown to promote proliferation
of cultured human coronary artery SMCs28 and
primary prostate epithelial cells.31 It has been reported
that the OPN-stimulated migration of vascular SMCs is mediated by
interaction of OPN with ß3
integrin32 and, likewise, that the growth-promoting effect
of OPN is mediated by
vß3
integrin.33 In the present study, we have also found
that OPN stimulates migration and enhances PDGF-BB–mediated DNA
synthesis of cultured rat aortic SMCs; thus, we examined the effect of
OPN on downstream events of the integrin signaling pathway. As shown in
Figures 4 and 5, our present data clearly indicate
that OPN and PDGF-BB synergistically activate FAK and ERK; this
finding seems to explain the OPN-induced enhancement of
PDGF-BB–mediated DNA synthesis observed in the present study.
We have previously reported that overexpression of the PDGF ß-receptor in SMCs is a causative element in the accelerated growth of diabetic SMCs.34 35 The report, together with our present findings, raises a possibility that diabetes-induced upregulation of the PDGF ß-receptor expression and OPN production in vascular SMCs synergistically augment their migration and proliferation and, thereby, facilitate the development of atherosclerosis in diabetes mellitus. Further study is necessary to examine this possibility.
|
Acknowledgments |
|---|
This study was supported by grants from the Ministry of Education, Science, Sports, and Culture of Japan (Nos. 11144206 and 11838002) and from the Sumitomo Foundation for Medicine and Welfare.
Received July 5, 1999; accepted September 28, 1999.
|
References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Andersen JL, Rasmussen LM, Ledet T. Diabetic macroangiopathy and atherosclerosis. Diabetes. 1996;45:S91–S94.
2. American Diabetes Association. Role of cardiovascular risk factors in prevention and treatment of macrovascular disease in diabetes. Diabetes Care. 1989;12:573–579.[Medline] [Order article via Infotrieve]
3.
van Belle E, Bauters C, Hubert E, Bodart JC, Abolmaali
K, Meurice T, McFadden EP, Lablanche JM, Bertrand ME.
Restenosis rates in diabetic patients. Circulation. 1997;96:1454–1460.
4. Ferrier TM. Radiologically demonstrable arterial calcification in diabetes mellitus. Aust Ann Med. 1964;13:222–228.
5. Christensen NJ. Muscle blood flow, measured by xenon113 and vascular calcifications in diabetics. Acta Med Scand. 1968;183:449–454.[Medline] [Order article via Infotrieve]
6. Butler WT. The nature and significance of osteopontin. Connect Tissue Res. 1989;23:123–136.[Medline] [Order article via Infotrieve]
7. Patarca R, Saavedra RA, Cantor H. Molecular and cellular basis of genetic resistance to bacterial infection: the role of the early T-lymphocyte activation-1/osteopontin gene. Crit Rev Immunol. 1993;13:225–246.[Medline] [Order article via Infotrieve]
8. Oates AJ, Barraclough R, Rudland PS. The role of osteopontin in tumorigenesis and metastasis. Invasion Metastasis. 1997;17:1–15.[Medline] [Order article via Infotrieve]
9. Yamate T, Kohri K, Umekawa T, Iguchi M, Kurita T. Osteopontin antisense oligonucleotide inhibits adhesion of calcium oxalate crystals in Madin-Darby canine kidney cell. J Urol. 1998;160:1506–1512.[Medline] [Order article via Infotrieve]
10. Liaw L, Birk DE, Ballas CB, Whitsitt JS, Davidson JM, Hogan BL. Altered wound healing in mice lacking a functional osteopontin gene (suppl 1). J Clin Invest. 1998;101:1468–1478.[Medline] [Order article via Infotrieve]
11. Shanahan CM, Cary NR, Metcalfe JC, Weissberg PL. High expression of genes for calcification-regulating proteins in human atherosclerotic plaques. J Clin Invest. 1994;93:2393–2402.
12.
Liaw L, Lombardi DM, Almeida MM, Schwartz SM, deBlois
D, Giachelli CM. Neutralizing antibodies directed against osteopontin
inhibit rat carotid neointimal thickening after
endothelial denudation. Arterioscler Thromb Vasc
Biol. 1997;17:188–193.
13.
Wada T, McKee MD, Steitz S, Giachelli CM. Calcification
of vascular smooth muscle cell cultures: inhibition by osteopontin.
Circ Res. 1999;84:166–178.
14. Takemoto M, Yokote K, Yamazaki M, Ridall AL, Butler WT, Matsumoto T, Tamura K, Saito Y, Mori S. Enhanced expression of osteopontin by high glucose in cultured rat aortic smooth muscle cells. Biochem Biophys Res Commun. 1999;258:722–726.[Medline] [Order article via Infotrieve]
15. Katagiri YU, Murakami M, Mori K, Iizuka J, Hara T, Tanaka K, Jia WY, Chambers AF, Uede T. Non-RGD domains of osteopontin promote cell adhesion without involving alpha v integrins. J Cell Biochem. 1996;62:123–131.[Medline] [Order article via Infotrieve]
16. Kanzaki T, Ishikawa Y, Morisaki N, Shirai K, Saito Y, Yoshida S. Abnormal metabolism of polyunsaturated fatty acid and phospholipids in diabetic glomeruli. Lipids. 1987;22:704–710.[Medline] [Order article via Infotrieve]
17. Wakamatsu K, Ghazizadeh M, Ishizaki M, Fukuda Y, Yamanaka N. Optimizing collagen antigen unmasking in paraffin-embedded tissues. Histochem J. 1997;29:65–72.[Medline] [Order article via Infotrieve]
18. Kanzaki T, Shiina R, Saito Y, Zardi L, Morisaki N. Transforming growth factor-ß receptor and fibronectin expressions in aortic smooth muscle cells in diabetic rats. Diabetologia. 1997;40:383–391.[Medline] [Order article via Infotrieve]
19. Bills CE, Eisenberg H, Pallante SL. Complexes of organic acids with calcium phosphate: the von Kossa stain as a clue to the composition of bone mineral. Johns Hopkins Med J. 1974;128:194–207.[Medline] [Order article via Infotrieve]
20. Morisaki N, Takahashi K, Shiina R, Zenibayashi M, Otabe M, Yoshida S, Saito Y. Platelet-derived growth factor is a potent stimulator of expression of intercellular adhesion molecule-1 in human arterial smooth muscle cells. Biochem Biophys Res Commun. 1994;200:612–618.[Medline] [Order article via Infotrieve]
21.
Yokote K, Mori S, Siegbahn A, Rönnstrand L,
Wernstedt C. Heldin CH, Claesson-Welsh L. Structural determinants in
the platelet-derived growth factor ß-receptor implicated in
modulation of chemotaxis. J Biol Chem. 1996;271:5101–5111.
22.
Mori S, Claesson-Welsh L, Heldin CH. Identification of
a hydrophobic region in the carboxyl terminus of the
platelet-derived growth factor ß-receptor which is important
for ligand-mediated endocytosis. J Biol Chem. 1991;266:21158–21164.
23. Saito Y, Mori S, Yokote Y, Kanzaki T, Saito Y, Morisaki N. Phosphatidylinositol 3-kinase activity is required for the activation process of focal adhesion kinase by platelet-derived growth factor. Biochem Biophys Res Commun. 1996;224:23–26.[Medline] [Order article via Infotrieve]
24.
Matsumoto T, Yokote K, Tamura K, Takemoto M, Ueno H,
Saito Y, Mori S. Platelet-derived growth factor activates
p38 mitogen-activated protein kinase through a Ras-dependent
pathway that is important for actin reorganization and cell migration.
J Biol Chem. 1999;274:13954–13960.
25. Denhardt DT, Noda M. Osteopontin expression and function: role in bone remodeling. J Cell Biochem. 1998;31:92–102.
26.
Towler DA, Bidder M, Latifi T, Coleman T, Semenkovich
CF. Diet-induced diabetes activates an osteogenic gene
regulatory program in the aortas of low density lipoprotein
receptor-deficient mice. J Biol Chem. 1998;273:30427–30434.
27.
Fischer JW, Tschope C, Reinecke A, Giachelli CM, Unger
T. Upregulation of osteopontin expression in renal cortex of
streptozotocin-induced diabetic rats is mediated by bradykinin.
Diabetes. 1998;47:1512–1518.
28.
Panda D, Kundu GC, Lee BI, Peri A, Fohl D,
Chackalaparampil I, Mukherjee BB, Li XD, Mukherjee DC, Seides S,
Rosenberg J, Stark K, Mukherjee AB. Potential roles of osteopontin
and vß3 integrin in the development of coronary
artery restenosis after angioplasty. Proc Natl Acad Sci
U S A. 1997;94:9308–9313.
29. Giachelli CM, Lombardi D, Johnson RJ, Murry CE, Almeida M. Evidence for a role of osteopontin in macrophage infiltration in response to pathological stimuli in vivo. Am J Pathol. 1998;152:353–358.[Abstract]
30.
Faccio R, Grano M, Colucci S, Zallone AZ, Quaranta V,
Pelletier AJ. Activation of vß3 integrin on human
osteoclast-like cells stimulates adhesion and migration in response to
osteopontin. Biochem Biophys Res Commun. 1998;249:522–525.[Medline]
[Order article via Infotrieve]
31. Elgavish A, Prince C, Chang PL, Lloyd K, Lindsey R, Reed R. Osteopontin stimulates a subpopulation of quiescent human prostate epithelial cells with high proliferative potential to divide in vitro. Prostate. 1998;35:83–94.[Medline] [Order article via Infotrieve]
32.
Srivatsa SS, Fitzpatrick LA, Tsao PW, Reilly TM, Holmes
DR Jr, Schwartz RS, Mousa SA. Selective vß3 integrin blockade
potently limits neointimal hyperplasia and lumen
stenosis following deep coronary arterial
stent injury: evidence for the functional importance of integrin
vß3 and osteopontin expression during neointima
formation. Cardiovasc Res. 1997;36:408–428.
33. Yue TL, McKenna PJ, Ohlstein EH, Farach-Carson MC, Butler WT, Johanson K, McDevitt P, Feuerstein GZ, Stadel JM. Osteopontin-stimulated vascular smooth muscle cell migration is mediated by ß3 integrin. Exp Cell Res. 1994;214:459–464.[Medline] [Order article via Infotrieve]
34. Kawano M, Koshikawa T, Kanzaki T, Morisaki N, Saito Y, Yoshida S. Diabetes mellitus induces accelerated growth of aortic smooth muscle cells: association with overexpression of PDGF ß-receptors. Eur J Clin Invest. 1993;23:84–90.[Medline] [Order article via Infotrieve]
35. Tamura K, Yokote K, Takemoto M, Matsumoto T, Ishisaki A, Funa K, Saito Y, Mori S. Fibronectin stimulates transcription of the platelet-derived growth factor ß-receptor in cultured rat aortic smooth muscle cells. Biochem Biophys Res Commun. 1998;251:677–680.[Medline] [Order article via Infotrieve]
This article has been cited by other articles:
 |
|
 |
|
  M. Koshikawa, K. Aizawa, H. Kasai, A. Izawa, T. Tomita, S. Kumazaki, H. Tsutsui, J. Koyama, S. Shimodaira, M. Takahashi, et al. Elevated Osteopontin Levels in Patients With Peripheral Arterial Disease Angiology, February 1, 2009; 60(1): 42 - 45. [Abstract] [PDF]
|
|
|
|
 |
|
 S. Jalvy, M.-A. Renault, L. L. S. Leen, I. Belloc, J. Bonnet, A.-P. Gadeau, and C. Desgranges Autocrine expression of osteopontin contributes to PDGF-mediated arterial smooth muscle cell migration Cardiovasc Res, September 1, 2007; 75(4): 738 - 747. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Bouvet, W. Peeters, S. Moreau, D. deBlois, and P. Moreau A new rat model of diabetic macrovascular complication Cardiovasc Res, February 1, 2007; 73(3): 504 - 511. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Andre, T. Stoeger, S. Takenaka, M. Bahnweg, B. Ritter, E. Karg, B. Lentner, C. Reinhard, H. Schulz, and M. Wjst Inhalation of ultrafine carbon particles triggers biphasic pro-inflammatory response in the mouse lung Eur. Respir. J., August 1, 2006; 28(2): 275 - 285. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Homo-Delarche, S. Calderari, J.-C. Irminger, M.-N. Gangnerau, J. Coulaud, K. Rickenbach, M. Dolz, P. Halban, B. Portha, and P. Serradas Islet Inflammation and Fibrosis in a Spontaneous Model of Type 2 Diabetes, the GK Rat Diabetes, June 1, 2006; 55(6): 1625 - 1633. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Kato, Y. Momiyama, R. Ohmori, N. Tanaka, H. Taniguchi, K. Arakawa, M. Kusuhara, H. Nakamura, and F. Ohsuzu High Plasma Levels of Osteopontin in Patients With Restenosis After Percutaneous Coronary Intervention Arterioscler. Thromb. Vasc. Biol., January 1, 2006; 26(1): e1 - e2. [Full Text] [PDF]
|
|
|
|
|
|
L. Li, A.-C. Jonsson-Rylander, K. Abe, and Z. Zukowska Chronic Stress Induces Rapid Occlusion of Angioplasty-Injured Rat Carotid Artery by Activating Neuropeptide Y and Its Y1 Receptors Arterioscler. Thromb. Vasc. Biol., October 1, 2005; 25(10): 2075 - 2080. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Fuster, P. R. Moreno, Z. A. Fayad, R. Corti, and J. J. Badimon Atherothrombosis and High-Risk Plaque: Part I: Evolving Concepts J. Am. Coll. Cardiol., September 20, 2005; 46(6): 937 - 954. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Y. Speer, Y.-C. Chien, M. Quan, H.-Y. Yang, H. Vali, M. D. McKee, and C. M. Giachelli Smooth muscle cells deficient in osteopontin have enhanced susceptibility to calcification in vitro Cardiovasc Res, May 1, 2005; 66(2): 324 - 333. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Sahai, P. Malladi, H. Melin-Aldana, R. M. Green, and P. F. Whitington Upregulation of osteopontin expression is involved in the development of nonalcoholic steatohepatitis in a dietary murine model Am J Physiol Gastrointest Liver Physiol, July 1, 2004; 287(1): G264 - G273. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Vattikuti and D. A. Towler Osteogenic regulation of vascular calcification: an early perspective Am J Physiol Endocrinol Metab, May 1, 2004; 286(5): E686 - E696. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Kawamura, K. Yokote, S. Asaumi, K. Kobayashi, M. Fujimoto, Y. Maezawa, Y. Saito, and S. Mori High Glucose-Induced Upregulation of Osteopontin Is Mediated via Rho/Rho Kinase Pathway in Cultured Rat Aortic Smooth Muscle Cells Arterioscler. Thromb. Vasc. Biol., February 1, 2004; 24(2): 276 - 281. [Abstract] [Full Text]
|
|
|
|
 |
|
 J.-S. Shao, S.-L. Cheng, N. Charlton-Kachigian, A. P. Loewy, and D. A. Towler Teriparatide (Human Parathyroid Hormone (1-34)) Inhibits Osteogenic Vascular Calcification in Diabetic Low Density Lipoprotein Receptor-deficient Mice J. Biol. Chem., December 12, 2003; 278(50): 50195 - 50202. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Y. Speer, M. D. McKee, R. E. Guldberg, L. Liaw, H.-Y. Yang, E. Tung, G. Karsenty, and C. M. Giachelli Inactivation of the Osteopontin Gene Enhances Vascular Calcification of Matrix Gla Protein-deficient Mice: Evidence for Osteopontin as an Inducible Inhibitor of Vascular Calcification In Vivo J. Exp. Med., October 21, 2002; 196(8): 1047 - 1055. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Li, S. Oparil, S. S. Kelpke, Y.-F. Chen, and J. A. Thompson Fibroblast Growth Factor Receptor-1 Signaling Induces Osteopontin Expression and Vascular Smooth Muscle Cell-Dependent Adventitial Fibroblast Migration In Vitro Circulation, August 13, 2002; 106(7): 854 - 859. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Isoda, K. Nishikawa, Y. Kamezawa, M. Yoshida, M. Kusuhara, M. Moroi, N. Tada, and F. Ohsuzu Osteopontin Plays an Important Role in the Development of Medial Thickening and Neointimal Formation Circ. Res., July 12, 2002; 91(1): 77 - 82. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. P. Sodhi, S. A. Phadke, D. Batlle, and A. Sahai Hypoxia Stimulates Osteopontin Expression and Proliferation of Cultured Vascular Smooth Muscle Cells: Potentiation by High Glucose Diabetes, June 1, 2001; 50(6): 1482 - 1490. [Abstract] [Full Text]
|
|
|
|
 |
|
 F. Takahashi, K. Takahashi, T. Okazaki, K. Maeda, H. Ienaga, M. Maeda, S. Kon, T. Uede, and Y. Fukuchi Role of Osteopontin in the Pathogenesis of Bleomycin-Induced Pulmonary Fibrosis Am. J. Respir. Cell Mol. Biol., March 1, 2001; 24(3): 264 - 271. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||