Thrombosis |
From the Department of Medicine (Y.C., D.J.S.), The University of Vermont (Burlington); and Department of Medicine (J.J.B.), Washington University, St Louis, Mo.
Correspondence to David J. Schneider, MD, College of Medicine, University of Vermont, 208 South Park Dr, Suite 2, Colchester, VT 05446. E-mail djschnei{at}zoo.uvm.edu
| Abstract |
|---|
|
|
|---|
Key Words: Diabetes mellitus PAI-1 free fatty acids Sp1
| Introduction |
|---|
|
|
|---|
Both FFAs and their metabolites, particularly long-chain acyl-CoA, have been shown to regulate the expression of genes through binding to or modification of transcription factors.8 9 10 11 We postulated that the induction of PAI-1 by FFAs is secondary to a direct or an indirect effect of FFAs on the expression and/or activation of a transcription factor or factors that bind to a cis-acting element in the 5' flanking region of the PAI-1 gene. The present study was designed to identify the cis-acting element or elements.
| Methods |
|---|
|
|
|---|
The sodium salts of FFAs (Sigma Chemical Co) were dissolved in water at 37°C and added dropwise to 1% fatty acidfree bovine serum albumin (FAF-BSA; Sigma Chemical Co) in DME-F12. We determined that 3% BSA in DME-F12 contains 0.45 mmol/L FFAs (Wako NEFA C kits; Biochemical Diagnostics). All solutions were adjusted to pH 7.4 and filtered through a 0.22-µm filter.
Eighty percent confluent HepG2 cells were preincubated in serum-free DME-F12 for 4 to 8 hours. Subsequently, cells were exposed to DME-F12 with FAF-BSA, DME-F12 with FAF-BSA to which selected fatty acids were added, or DME-F12 with BSA. Conditioned media were collected after selected intervals and assayed for PAI-1 by ELISA (Tintelize; Biopool). Total cellular RNA was isolated conventionally with TRIzol Reagent (GIBCO BRL). Northern blotting was performed as previously described with 10 µg RNA.12
Construction of PAI-1 Promoter-Luciferase
Reporter Plasmids
A 1387-bp KpnI fragment containing
1313 bp of 5'-flanking (-1313) and 74 bp of the untranslated first
exon (+74) of human PAI-1 DNA was isolated from the plasmid PAI-CAT
1313,13 inserted
into the unique KpnI site of the promoter-less
luciferase reporter plasmid pGL3-Basic (Promega), and referred to as
PAI-LUC 1313. A 927-bp XhoI fragment (from -853 to
+74 in human PAI-1 DNA) was inserted into the XhoI
site of pGL3-Basic and referred to as PAI- LUC 853. Polymerase chain
reaction was used to amplify a 684-bp fragment that encompassed -610
to +74 (5' primer, 5'-cta ggt acc aga cca aga gtc ctc tgt tg-3'; 3'
primer, 5'-aat gga tcc gaa ttc agc tgc tgg agg-3'). This fragment was
inserted into pGL3-Basic and referred to as PAI-LUC 610. A 402-bp
HindIII fragment containing -328 to +74 bp of the
PAI-1 gene was inserted into pGL3-Basic and referred to as PAI-LUC 328.
A 72-bp ApaI fragment (-599 to -528) containing a
potential fatty acid response
element14 was
deleted from PAI-LUC 1313 to form PAI-LUC 1313-72. To generate
PAI-LUC 328+72, the 72-bp ApaI fragment was inserted
upstream of the minimal promoter (PAI-LUC 328) in the sense
orientation. The 72+pGL3-Promoter was generated by inserting the 72-bp
KpnI/XhoI fragment upstream of the
simian virus (SV)40 promoter in pGL3-Promoter (Promega).
Transient transfection of HepG2 cells was performed according to the calcium-phosphate precipitation method as previously described.13 Transfections were performed with 12 µg reporter plasmid and 600 ng pRL-cytomegalovirus (CMV), a plasmid with Renilla luciferase gene downstream of the CMV promoter to control for transfection efficiency. Luciferase activity was detected in cell extracts with the Passive Lysis Buffer in the Dual-Luciferase Reporter Assay System (Promega). Luciferase activities were determined with the Dual-Luciferase Reporter Assay with a DLReady luminometer (Turner Designs Instrument).
DNase I footprinting was performed with -618 to -487 of the PAI-1 gene in accordance with the Maxam method.15 Protein extract (100 µg) was used in a 50-µL reaction that contained 10 mmol/L Tris, pH 7.5, 50 mmol/L NaCl, 1 mmol/L EDTA, 1% Ficoll, and 1 µg poly(dI/dC)·poly(dI/dC).
Electrophoretic Mobility Shift Assays
Proteins were extracted from HepG2 cells exposed to
control media (DME-F12 with 1% FAF-BSA) and to 0.75 mmol/L capric
acid in DME-F12 with 1% FAF-BSA according to the freeze-thaw method
described by Ladias et
al.16
Oligonucleotides of PAI-1 5' flanking DNA and Sp1 consensus sequences17 that were used in electrophoretic mobility shift assays (EMSAs) were synthesized at Life Technology (GIBCO BRL) and include 29 bp, 5'-gcatgccctgtggctgttgggctgggccc-3' and 5'-gggcccagcccaacagccacagggcatgc-3'; 15 bp, 5'-tggctgttgggctgg-3' and 5'-ccagcccaacagcca-3'; 9 bp, 5'-tggctgttg-3' and 5'-caacagcca-3'; 23 bp, 5'-cacgtggctggctgcatgccctg-3' and 5'-cagggcatgcagcca-gccacgtg-3'; 18 bp, 5'-cacgtggctggctgcatg-3' and 5'-catgcagcca-gccacgtg-3'; 13 bp, 5'-cacgtggctggct-3' and 5'-agccagccacgtg-3'; 10 bp, 5'-gcatgccctg-3' and 5'-cagggcatgc-3'; C13, 5'-tggctgctggctg-3' and 5'-cagccagcagcca-3'; M13, 5'-taactgctaactg-3' and 5'-cagttagcagtta-3'; 30 bp, 5'-aagtcctagacagacaaaacctagacaatc-3' and 5'-gattgtctaggttttgtctgtctaggactt-3'; Sp1, 5'-acaattgggctgggcctaat-3' and 5'-attaggcccagcccaattgt-3' and 5'-attcgatcggggcggggcgagc-3' and 5'-gctcgccccgccccgatcgaat-3' (Promega); and 24 bp, 5'-tcacagaacatgtctctatcgtaa-3' and 5'-ttacgatagagagatgttcagtga-3' (control).
Double-strand oligonucleotides were
end-labeled with (
-32P)-ATP and T4
polynucleotide kinase. Unincorporated
nucleotides were removed with a NucTrap column
(Stratagene). EMSAs were performed as previously
described.18
Statistical Analysis
Results are given as mean±SEM. Differences between 2
groups were identified with a Students t test. For
multiple groups, 1-way ANOVA and Student-Newman-Keuls tests were used
to identify differences. Significance was defined as
P<0.05.
| Results |
|---|
|
|
|---|
|
FFAs and Expression of PAI-1 mRNA
Exposure to 0.75 mmol/L capric acid led to a
2±0.02-fold increase in 3.2-kb PAI-1 mRNA and a 2.8±0.1-fold increase
in 2.2-kb PAI-1 mRNA (n=3, P<0.001). Maximal
increases were seen 16 to 48 hours after
exposure.
FFAs and Luciferase Activity
HepG2 cells were transfected with PAI-LUC reporter
constructs
(Figure 2
). A 1.7-fold increase in luciferase activity
(corrected for transfection efficiency) was seen in extracts from cells
transfected with PAI-LUC 1313, 853, or 610 after exposure to 0.75
mmol/L albumin-bound FFAs (n=6 for each). Comparable results
were seen with capric acid and with 3% BSA binding a
physiological mixture of FFAs (fold induction with
3% BSA, 1.36±0.02 for PAI-LUC 1313 and 1.35±0.03 for PAI-LUC 610,
n=6 for each, P<0.001).
|
After the deletion of a 72-bp ApaI fragment (-599 to -528, PAI-LUC 1313-72), neither 3% BSA nor capric acid augmented luciferase activity. FFA-induced increased luciferase activity was seen when the 72-bp segment was inserted upstream of the minimal promoter (PAI-LUC 328+72, 1.43±0.04-fold increase with 3% BSA, n=6, P<0.001). Increased FFA-induced luciferase activity was also seen when the 72-bp fragment was inserted upstream of the SV40 promoter in pGL3-Promoter (1.29±0.01- fold, n=8, P<0.001). Accordingly, the 72-bp segment, from -599 to -528 of the PAI-1 gene, contains the fatty acid response region.
Interaction Between Extracted Proteins and
Fatty Acid Response Region
DNase I footprinting was performed with a 130-bp
end-labeled fragment spanning -618 to -488 in the PAI-1 promoter
region. This segment contains the 72-bp region with the implicated
fatty acid response element in addition to a 20-bp sequence upstream
and a 40-bp sequence downstream. The fragment was exposed to extracts
from cells treated with 0.75 mmol/L capric acid. Two regions
(referred to as A and B) in the end-labeled 130-bp fragment were
protected from DNase I digestion
(Figure 3
). Footprinted site A corresponds to
nucleotides -524 to -558. Footprinted site B
corresponds to nucleotides -559 to
-590.
|
Interaction of DNA With Proteins Extracted From
HepG2 Cells
One fragment that encompasses the entire footprinted A
region, 2 overlapping fragments (29 and 23 bp) that encompass the
footprinted A region, and 1 fragment that encompasses the footprinted B
region (30 bp) were used for gel mobility shift analysis. These
fragments exhibited specific binding to proteins extracted from HepG2
cells (n=5 for each;
Figure 4
). Results with the fragment that encompasses the
entire footprinted region A were similar to those seen with the 29-bp
fragment (data not shown). Although 3 DNA/protein complexes were
identified with the selected fragments, only binding of complex I was
consistently inhibited with self "cold"
(non-labeled). No inhibition was seen when a 24-bp unrelated fragment
(100-fold excess) was used as competitor
(Figure 4
).
|
FFAs and Binding of Proteins Extracted From
HepG2 Cells to Fatty Acid Response Region
Proteins extracted from cells exposed to control
conditions (FAF-BSA) and to FFAs were exposed to
oligonucleotides in EMSAs. Induction of complex I was
seen when the 29- or 30-bp probe was exposed to proteins from cells
treated with FFAs. Increased binding was seen 5 to 120 minutes after
exposure to FFAs (n=3 for each).
Identification of Fatty Acid Response
Element
DNA sequence analysis of the fatty acid
responsive region identified 4 repeats of the sequence
5'-TG(G/C)12CTG-3' in the 23- and 29-bp
fragments. The EMSAs demonstrated that the fatty acid responsive
element is common to the 29- and 23-bp fragments. Progressively smaller
components of the 29- and 23-bp fragments were used to delineate
further the fatty acid response element. Complex I was seen with each
fragment, including those with a single copy of the implicated fatty
acid response element
(Figure 5
). In addition, EMSAs were performed with a 13-bp
oligonucleotide that contained the sequence 5'-TGGCTG C
TGGCTG-3'. This oligonucleotide contains 2 repeats of
the implicated fatty acid response element with 1 spacer (C). The same
shifted pattern was seen when the oligonucleotide was
incubated with proteins extracted from HepG2 cells. Mutation of
"GG" to "AA" abolished the binding when the probe
5'-TAACTG C TAACTG-3' was used in the
EMSAs.
|
Involvement of a Transcription Factor That
Binds to Sp1 Consensus Sites
Footprinted region A contains 4 repeats of the
implicated fatty acid response element. The sequence is similar to the
Sp1 binding sequence (T/G)(G/A)GGC(T/G)G(G/A)(G/A)(C/T). In addition,
footprinted region B contains a 10-bp segment that has 70% identity
with the Sp1 consensus site. Thus, EMSAs were performed with Sp1
consensus oligonucleotides and demonstrated a
DNA-protein complex with an electrophoretic mobility pattern similar to
that seen with complex I
(Figure 6
). Binding of extracted protein to complex I with
the 29- or 23-bp fragment was inhibited with a 100-fold molar excess of
unlabeled Sp1 consensus DNA
(Figure 6
). Inhibition was seen also when extracted proteins
were exposed to unlabeled 29- or 23-bp fragments before exposure to
radiolabeled Sp1 consensus
oligonucleotides.
|
The 30-bp fragment inhibited binding of proteins to complex
I when competition EMSAs were performed with the 29-bp, 23-bp, or Sp1
consensus
(Figure 6
). Similarly, unlabeled 29-bp, 23-bp, and Sp1
consensus oligonucleotides inhibited the binding of
proteins to the radiolabeled 30-bp fragment. Thus, the results of the
competition EMSAs suggest that a protein that can bind to Sp1 consensus
DNA increases transcription of the PAI-1 gene after exposure of the
HepG2 cells to FFAs.
| Discussion |
|---|
|
|
|---|
In the present study, we found that a physiological mixture of fatty acids associated with BSA as well as 2 medium-chain length fatty acids (capric acid and lauric acid) and a long-chain polyunsaturated fatty acid (linoleic acid) increase the expression of PAI-1 in HepG2 cells. Our results are consistent with those of others that demonstrate increased expression of PAI-1 by FFAs of varying chain length and saturation.4 23 24 Thus, the effect of fatty acids on the expression of PAI-1 appears to be related to the prevailing concentration of diverse FFAs rather than to the concentration of a specific fatty acid.
We used chimeric genes that contain firefly luciferase as a reporter downstream of selected segments of 5' flanking DNA from the human PAI-1 gene to identify the mechanism by which fatty acids augment the expression of PAI-1. Deletion analysis of the PAI-1 promoter region indicated that the fatty acid response region is located between -599 and -528 in the PAI-1 5' flanking region. The implicated fatty acid response element is distinct from the VLDL response element identified by Erikssen et al.6 These results suggest that FFAs augment the expression of PAI-1 independent of effects mediated by triglycerides or lipoproteins.
DNA sequence analysis demonstrated that the implicated fatty acid response sequence was repeated 4 times in the footprinted region A. The footprinted region B contains a sequence "TGTCTG" in the complementary strand that is 83% identical to the implicated fatty acid response element. Sp1 recognition sequences are functional in either orientation17 Thus, despite the asymmetry of Sp1-binding sites, competition between the 30-bp fragment and the 29- or 23-bp fragment would be expected. Further, the increased binding of proteins to the fatty acid response element seen within 5 minutes after exposure of the cells to FFAs suggests activation rather than induction of a transcription factor. Accordingly, increased expression of PAI-1 after exposure of the HepG2 cells to FFAs is secondary to activation of a transcription factor that binds to a repeated element in the 5' flanking region of the PAI-1 gene.
Three fatty acid response regions have been described previously.10 25 26 A cis-linked fatty acid response element was localized to a 140-bp (-80 to -220) region within the S14 proximal promoter.10 25 26 In the pyruvate kinase gene, a fatty acid response region was identified that overlaps the glucose/insulin response element. These results suggest that fatty acids inhibit the pyruvate kinase transcription by interfering with glucose and insulin induction of the gene.25 Specific binding of trans-acting proteins to the response regions was not defined in these studies. The third fatty acid responsive region was identified in the stearoyl-CoA desaturase gene 1 (SCD 1). Proteins from HepG2 cells and 3T3-L1 cells have been shown to bind specifically to a 60- bp region in the SCD1 promoter.26 Thus, the present results constitute the first report of a Sp1 consensus site that mediates the effects of fatty acids.
Sp1-like proteins are a family of proteins that contain 3 highly conserved C-terminal zinc finger domains and bind to GC-rich sequences.17 27 Consistent with our observation, Sp1-responsive promoters usually contain multiple recognition sites, although a single binding site appears to be sufficient for a promoter to be stimulated by Sp1.17
Previous studies have demonstrated that Sp1 sites contribute to transcriptional activation of the rat PAI-1 gene.28 Proteins extracted from HTC rat hepatoma cells bind to Sp1 and CTF/NF-1like binding sites in the rat PAI-1 regulatory region.28 An Sp1-binding site identified in the rat PAI-1 gene corresponds to the binding sites identified in footprint A in the present study. The activation of Sp1 in a different region of the PAI-1 gene has been implicated in glucose-mediated increased expression of PAI-1.29 In studies with rat aortic smooth muscle cells, glucose has been shown to release a transcriptional repressor from Sp1 complexes. By contrast, we found that FFAs activate a transcription factor that binds to Sp1 consensus sites. Thus, the increased expression of PAI-1 seen in subjects with diabetes appears to be secondary, at least in part, to increased transcription of PAI-1 induced by Sp1-binding sites and mediated by increased concentrations of both glucose and FFAs.
In summary, we have found that fatty acids augment the expression of PAI-1 in a human hepatoma cell line (HepG2). FFAs activate a transcription factor that binds to the sequence 5'-TG(G/C)12CTG-3' in the 5' flanking DNA of the human PAI-1 gene. This sequence is nearly identical to the DNA-binding site for Sp1. Further characterization of the transcription factor responsible for FFA-induced augmented expression of PAI-1 and its interaction with other transcription factors may provide a therapeutic target for modification of the expression of PAI-1 in diabetic subjects.
| Acknowledgments |
|---|
Received July 19, 2000; accepted September 5, 2000.
| References |
|---|
|
|
|---|
2.
Auwerx
J, Bouillon R, Collen D, Geboers J. Tissue-type plasminogen
activator antigen and plasminogen
activator inhibitior in diabetes mellitus.
Arteriosclerosis. 1998;8:6872.
3. Calles-Escandon J, Mirza SA, Sobel BE, Schneider DJ. Induction of hyperinsulinemia combined with hyperglycemia and hypertriglyceridemia increases plasminogen activator inhibitor 1 in blood in normal human subjects. Diabetes. 1998;47:290293.[Abstract]
4. Banfi C, Risé P, Mussoni l., Galli C, Tremoli E. Linoleic acid enhances the secretion of plasminogen activator inhibitor type 1 by HepG2 cells. J Lipid Res. 1997;38:860869.[Abstract]
5.
Sironi
L, Mussoni l., Prati L, Baldassarre D, Camera M, Banfi C, Tremoli E.
Plasminogen activator inhibitor
type-1 synthesis and mRNA expression in HepG2 cells are regulated by
VLDL. Arterioscler Thromb Vasc Biol. 1996;16:8996.
6.
Eriksson
P, Nilsson L, Karpe F, Hamsten A. Very-low-density lipoprotein response
element in the promoter region of the human plasminogen
activator inhibitor-1 gene implicated in the
impaired fibrinolysis of
hypertriglyceridemia. Arterioscler
Thromb Vasc Biol. 1998;18:2026.
7. Schneider DJ, Sobel BE. Synergistic augmentation of expression of plasminogen activator inhibitor type-1 induced by insulin, very-low-density lipoproteins, and fatty acids. Coron Artery Dis. 1996;7:813817.[Medline] [Order article via Infotrieve]
8. Hertz R, Magenheim J, Berman I, Bar-Tana J. Fatty acyl-CoA thioesters are ligands of hepatic nuclear factor-4alpha. Nature. 1998;392:512516.[Medline] [Order article via Infotrieve]
9. Amri EZ, Ailhaud G, Grimaldi PA. Fatty acids as signal transducing molecules: involvement in the differentiation of preadipose to adipose cells. J Lipid Res. 1994;35:930937.[Abstract]
10.
Jump
DB, Clarke SD, MacDougald O, Thelen A. Polyunsaturated fatty acids
inhibit S14 gene transcription in rat liver and cultured
hepatocytes. Proc Natl Acad Sci
U S A. 1993;90:84548458.
11.
Keller
H, Dreyer C, Medin J, Mahfoudi A, Ozato K, Wahli W. Fatty acids and
retinoids control lipid metabolism through activation of
peroxisome proliferator-activated receptor-retinoid X receptor
heterodimers. Proc Natl Acad Sci
U S A. 1993;90:21602164.
12.
Schneider
DJ, Sobel BE. Augmentation of synthesis of plasminogen
activator inhibitor type 1 by insulin and
insulin-like growth factor type I: implications for vascular disease in
hyperinsulinemic states [published erratum appears in
Proc Natl Acad Sci
U S A. 1992;89:1148].
Proc Natl Acad Sci
U S A.
1991;88:99599963.
13.
Westerhausen
DRJ, Hopkins WE, Billadello JJ. Multiple transforming growth
factor-beta-inducible elements regulate expression of the
plasminogen activator inhibitor
type-1 gene in Hep G2 cells. J Biol Chem. 1991;266:10921100.
14. Sloots JA, Aitchison JD, Rachubinski RA. Glucose-responsive and oleic acid-responsive elements in the gene encoding the peroxisomal trifunctional enzyme of Candida tropicalis. Gene. 1991;105:129134.[Medline] [Order article via Infotrieve]
15. Maxam AM, Gilbert W. A new method for sequencing DNA. 1977 [classic article]. Biotechnology. 1992;24:99103.[Medline] [Order article via Infotrieve]
16.
Ladias
JA, Hadzopoulou-Cladaras M, Kardassis D, Cardot P, Cheng J, Zannis V,
Cladaras C. Transcriptional regulation of human apolipoprotein genes
apoB, apoCIII, and apoAII by members of the steroid hormone receptor
superfamily HNF-4, ARP-1, EAR-2, and EAR-3. J Biol
Chem. 1992;267:1584915860.
17. Kadonaga JT, Carner KR, Masiarz FR, Tjian R. Isolation of cDNA encoding transcription factor Sp1 and functional analysis of the DNA binding domain. Cell. 1987;51:10791090.[Medline] [Order article via Infotrieve]
18.
Fried
M, Crothers DM. Equilibria and kinetics of lac repressor-operator
interactions by polyacrylamide gel electrophoresis.
Nucleic Acids Res. 1981;9:65056525.
19. Mehta J, Mehta P, Lawson D, Saldeen T. Plasma tissue plasminogen activator inhibitor levels in coronary artery disease: correlation with age and serum triglyceride concentrations. J Am Coll Cardiol. 1987;9:263268.[Abstract]
20. Hamsten A, de Faire U, Walldius G, Dahlen G, Szamosi A, Landou C, Blomback M, Wiman B. Plasminogen activator inhibitor in plasma: risk factor for recurrent myocardial infarction. Lancet. 1987;2:39.[Medline] [Order article via Infotrieve]
21. Loskutoff DJ, van Aken BE, Seiffert D. Abnormalities in the fibrinolytic system of the vascular wall associated with atherosclerosis. Ann N Y Acad Sci. 1995;748:17783.
22. Aznar J, Estelles A. Role of plasminogen activator inhibitor type 1 in the pathogenesis of coronary artery diseases. Haemostasis. 1994;24:243251.[Medline] [Order article via Infotrieve]
23. Smith TJ, Piscatelli JJ, Andersen V, Wang HS, Lance P. n-Butyrate induces plasminogen activator inhibitor type 1 messenger RNA in cultured Hep G2 cells. Hepatology. 1996;23:866871.[Medline] [Order article via Infotrieve]
24. Kariko K, Rosenbaum H, Kuo A, Zurier RB, Barnathan ES. Stimulatory effect of unsaturated fatty acids on the level of plasminogen activator inhibitor-1 mRNA in cultured human endothelial cells. FEBS Lett. 1995;361:118122.[Medline] [Order article via Infotrieve]
25.
Liimatta
M, Towle HC, Clarke S, Jump DB. Dietary polyunsaturated fatty acids
interfere with the insulin/glucose activation of L-type pyruvate kinase
gene transcription. Mol Endocrinol. 1994;8:11471153.
26. Waters KM, Miller CW, Ntambi JM. Localization of a polyunsaturated fatty acid response region in stearoyl-CoA desaturase gene 1. Biochim Biophys Acta. 1997;1349:3342.[Medline] [Order article via Infotrieve]
27.
Hagen
G, Muller S, Beato M, Suske G. Cloning by recognition site screening of
two novel GT box binding proteins: a family of Sp1 related genes.
Nucleic Acids Res. 1992;20:55195525.
28.
Johnson
MR, Bruzdzinski CJ, Winograd SS, Gelehrter TD. Regulatory sequences and
protein-binding sites involved in the expression of the rat
plasminogen activator inhibitor-1
gene. J Biol Chem. 1992;267:1220212210.
29.
Chen
YQ, Su M, Walia RR, Hao Q, Covington JW, Vaughan DE. Sp1 sites mediate
activation of the plasminogen activator
inhibitor-1 promoter by glucose in vascular smooth muscle
cells. J Biol Chem. 1998;273:82258231.
This article has been cited by other articles:
![]() |
R. Serrano, J. Barrenetxe, J. Orbe, J. A. Rodriguez, N. Gallardo, C. Martinez, A. Andres, and J. A. Paramo Tissue-specific PAI-1 gene expression and glycosylation pattern in insulin-resistant old rats Am J Physiol Regulatory Integrative Comp Physiol, November 1, 2009; 297(5): R1563 - R1569. [Abstract] [Full Text] [PDF] |
||||
![]() |
K. Oishi, D. Uchida, N. Ohkura, R. Doi, N. Ishida, K. Kadota, and S. Horie Ketogenic Diet Disrupts the Circadian Clock and Increases Hypofibrinolytic Risk by Inducing Expression of Plasminogen Activator Inhibitor-1 Arterioscler Thromb Vasc Biol, October 1, 2009; 29(10): 1571 - 1577. [Abstract] [Full Text] [PDF] |
||||
![]() |
A. K. M. T. Zaman, C. J. French, D. J. Schneider, and B. E. Sobel A Profibrotic Effect of Plasminogen Activator Inhibitor Type-1 (PAI-1) in the Heart Experimental Biology and Medicine, March 1, 2009; 234(3): 246 - 254. [Abstract] [Full Text] [PDF] |
||||
![]() |
C. H. Byon, A. Javed, Q. Dai, J. C. Kappes, T. L. Clemens, V. M. Darley-Usmar, J. M. McDonald, and Y. Chen Oxidative Stress Induces Vascular Calcification through Modulation of the Osteogenic Transcription Factor Runx2 by AKT Signaling J. Biol. Chem., May 30, 2008; 283(22): 15319 - 15327. [Abstract] [Full Text] [PDF] |
||||
![]() |
A. M. Kanaya, C. Wassel Fyr, E. Vittinghoff, T. B. Harris, S. W. Park, B. H. Goodpaster, F. Tylavsky, and S. R. Cummings Adipocytokines and incident diabetes mellitus in older adults: the independent effect of plasminogen activator inhibitor 1. Arch Intern Med, February 13, 2006; 166(3): 350 - 356. [Abstract] [Full Text] [PDF] |
||||
![]() |
J. Dong, S. Fujii, H. Li, H. Nakabayashi, M. Sakai, S. Nishi, D. Goto, T. Furumoto, S. Imagawa, T. A.K.M. Zaman, et al. Interleukin-6 and Mevastatin Regulate Plasminogen Activator Inhibitor-1 Through CCAAT/Enhancer-Binding Protein-{delta} Arterioscler Thromb Vasc Biol, May 1, 2005; 25(5): 1078 - 1084. [Abstract] [Full Text] [PDF] |
||||
![]() |
H.-C. Chen and E. P. Feener MEK1,2 response element mediates angiotensin II--stimulated plasminogen activator inhibitor-1 promoter activation Blood, April 1, 2004; 103(7): 2636 - 2644. [Abstract] [Full Text] [PDF] |
||||
![]() |
K. E. Watson, A. L. Peters Harmel, and G. Matson Atherosclerosis in Type 2 Diabetes Mellitus: The Role of Insulin Resistance Journal of Cardiovascular Pharmacology and Therapeutics, December 1, 2003; 8(4): 253 - 260. [Abstract] [PDF] |
||||
![]() |
B. E. Sobel, D. J. Taatjes, and D. J. Schneider Intramural Plasminogen Activator Inhibitor Type-1 and Coronary Atherosclerosis Arterioscler Thromb Vasc Biol, November 1, 2003; 23(11): 1979 - 1989. [Abstract] [Full Text] [PDF] |
||||
![]() |
M.-C. Alessi, D. Bastelica, A. Mavri, P. Morange, B. Berthet, M. Grino, and I. Juhan-Vague Plasma PAI-1 Levels Are More Strongly Related to Liver Steatosis Than to Adipose Tissue Accumulation Arterioscler Thromb Vasc Biol, July 1, 2003; 23(7): 1262 - 1268. [Abstract] [Full Text] [PDF] |
||||
![]() |
W. T. Cefalu, D. J. Schneider, H. E. Carlson, P. Migdal, L. Gan Lim, M. P. Izon, A. Kapoor, A. Bell-Farrow, J. G. Terry, and B. E. Sobel Effect of Combination Glipizide GITS/Metformin on Fibrinolytic and Metabolic Parameters in Poorly Controlled Type 2 Diabetic Subjects Diabetes Care, December 1, 2002; 25(12): 2123 - 2128. [Abstract] [Full Text] [PDF] |
||||
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
ATVB Home | Subscriptions | Archives | Feedback | Authors | Help | AHA Journals Home | Search Copyright © 2000 American Heart Association, Inc. All rights reserved. Unauthorized use prohibited. |