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Platelet-derived growth factor is a cofactor in the induction of 1α(I) procollagen expression by transforming growth factor β1 in smooth muscle cells 

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Presentation on theme: "Platelet-derived growth factor is a cofactor in the induction of 1α(I) procollagen expression by transforming growth factor β1 in smooth muscle cells "— Presentation transcript:

1 Platelet-derived growth factor is a cofactor in the induction of 1α(I) procollagen expression by transforming growth factor β1 in smooth muscle cells  Brian G. Halloran, MD, Byung J. So, MD, B.Timothy Baxter, MD  Journal of Vascular Surgery  Volume 23, Issue 5, Pages (May 1996) DOI: /S (96) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

2 Fig. 1 Subconfluent human arterial SMC were treated with varying concentrations of rh TGF-β1 and cells counted every 24 hours. Results represent mean ± SD of triplicate samples. High-dose TGF-β1 (1.0 and 10.0 ng/ml) inhibited SMC proliferation at 48 hours (*, 1.0 and 10.0 ng/ml < control; p < 0.05), and at 72 and 96 hours (**, 1.0 and 10.0 ng/ml < control and 0.1 ng/ml; p < 0.05). Low-dose TGF-β1 exhibited a transient inhibitory effect at 48 hours compared with control (p < 0.05), but did not differ from control at 72 and 96 hours. Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

3 Fig. 2 Human arterial SMC proliferation as measured by tritiated thymidine incorporation was inhibited by all doses of TGF-β1 at 24 hours (*, p < 0.001), and by high-dose TGF-β1 (1.0 and 10.0 ng/ml) at 48 hours (**, p < 0.005). Data represent mean ± SD of triplicate samples. Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

4 Fig. 3 Subconfluent human arterial SMC were treated with 10 ng/ml TGF-β1 and RNA analyzed by RT-PCR for 1α(I) procollagen and glyceraldehyde-3-phosphate de-hydrogenase. Enhanced expression of type I collagen mRNA did not occur until 24 and 48 hours after TGF-β1 treatment. Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

5 Fig. 4 Southern blot analysis of RT-PCR products for type I collagen (COL1A1) and glyceraldehyde-3-phosphate dehydrogenase demonstrated dose-related increase in 1α(I) procollagen expression with increasing TGF-β1 levels at 24 hours. Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

6 Fig. 5 Arterial SMC were treated with TGF-β1 for 36 hours and total cellular RNA subjected to dot blot analysis and signal quantification. Membranes were sequentially probed for 1α(I) procollagen and glyceraldehyde-3-phosphate dehydrogenase for normalization. TGF-β1 stimulated 36% increase in type I collagen expression (3.75 ± 0.11) compared with control group (2.78 ± 0.32; *, p < 0.05). Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

7 Fig. 6 Human arterial SMC were treated with TGF-β1 (10 ng/ml), and RNA was reverse-transcribed and amplified (20 cycles PCR) by PDGF-A and glyceraldehyde-3-phosphate dehydrogenase oligonucleotide primers. Southern blot analysis of cDNA shows PDGF-A is constitutively expressed at very low levels and TGF-β1 induces early and sustained increase in PDGF-A mRNA. Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

8 Fig. 7 Subconfluent human arterial SMC were treated with TGF-β1 (10 ng/ml) and increasing doses of PDGF-AB neutralizing antibodies. RNA was extracted after 36 hours and analyzed by RT-PCR. TGF-β1 stimulation of type I procollagen expression is diminished with increasing doses of anti-PDGF-AB. Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

9 Fig. 8 Proposed role of PDGF-A in TGF-β1 mediated 1α(I) procollagen upregulation in human arterial SMC. TGF-β1 induces expression of PDGF-A mRNA and consequently increases PDGF-AA and PDGF-AB protein levels. Because neutralizing antibodies to PDGF attenuate TGF-β1 mediated procollagen expression, PDGF likely acts as co-factor to promote increased procollagen gene expression. Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

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