Limits...
Collagen inhibitory peptide R1R2 mediates vascular remodeling by decreasing inflammation and smooth muscle cell activation.

Lee TH, Sottile J, Chiang HY - PLoS ONE (2015)

Bottom Line: Morphometric analysis demonstrated that R1R2 reduced intima-media thickening compared to the control groups.This decrease was accompanied by decreased VCAM-1 and ICAM-1 expression.Our in vitro studies revealed that R1R2 attenuated SMC proliferation and migration, and also decreased monocyte adhesion and transendothelial migration through endothelial cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan.

ABSTRACT
The extracellular matrix (ECM) is a major constituent of the vessel wall. In addition to providing a structural scaffold, the ECM controls numerous cellular functions in both physiologic and pathologic settings. Vascular remodeling occurs after injury and is characterized by endothelial cell activation, inflammatory cell infiltration, phenotypic modulation of smooth muscle cells (SMCs), and augmented deposition of collagen-rich ECM. R1R2, a peptide derived from the bacterial adhesin SFS, with sequence homology to collagen, is known to inhibit collagen type I deposition in vitro by inhibiting the binding of fibronectin to collagen. However, the inhibitory effects of R1R2 during vascular remodeling have not been explored. We periadventitially delivered R1R2 to carotid arteries using pluronic gel in a vascular remodeling mouse model induced by blood flow cessation, and evaluated its effects on intima-media thickening, ECM deposition, SMC activation, and inflammatory cell infiltration. Morphometric analysis demonstrated that R1R2 reduced intima-media thickening compared to the control groups. R1R2 treatment also decreased collagen type I deposition in the vessel wall, and maintained SMC in the contractile phenotype. Interestingly, R1R2 dramatically reduced inflammatory cell infiltration into the vessel by ∼ 78%. This decrease was accompanied by decreased VCAM-1 and ICAM-1 expression. Our in vitro studies revealed that R1R2 attenuated SMC proliferation and migration, and also decreased monocyte adhesion and transendothelial migration through endothelial cells. Together, these data suggest that R1R2 attenuates vascular remodeling responses by decreasing inflammation and by modulating SMC proliferation and migration, and suggest that the R1R2 peptide may have therapeutic potential in treating occlusive vascular diseases.

Show MeSH

Related in: MedlinePlus

R1R2 reduces SMC proliferation and migration.(A) A7r5 SMC growth study over 96 hours. After serum starvation and treatment with R1R2 or scrambled peptides for 36 hours, cells were stimulated with 10% FBS over 96 hours. Cells were counted at the indicated time in the figure (n = 5). (B) A7r5 SMCs were pretreated with R1R2 or scramble peptide for 36 hours. SMCs underwent a scratch wound injury and were then stimulated with 10 ng/ml PDGF-BB for 30 hours. Representative images from the scratch wound assays are shown. (C) Quantitation of SMC migration in the scratch wound healing assay was performed by subtracting the cell-free area 30 hours after PDGF-BB stimulation from the cell-free area in the beginning (n = 5). * indicates p<0.05 and ** p< 0.01.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4326127&req=5

pone.0117356.g006: R1R2 reduces SMC proliferation and migration.(A) A7r5 SMC growth study over 96 hours. After serum starvation and treatment with R1R2 or scrambled peptides for 36 hours, cells were stimulated with 10% FBS over 96 hours. Cells were counted at the indicated time in the figure (n = 5). (B) A7r5 SMCs were pretreated with R1R2 or scramble peptide for 36 hours. SMCs underwent a scratch wound injury and were then stimulated with 10 ng/ml PDGF-BB for 30 hours. Representative images from the scratch wound assays are shown. (C) Quantitation of SMC migration in the scratch wound healing assay was performed by subtracting the cell-free area 30 hours after PDGF-BB stimulation from the cell-free area in the beginning (n = 5). * indicates p<0.05 and ** p< 0.01.

Mentions: SMC proliferation and migration contribute to intima and media thickening after arterial injury [44,45]. Therefore, we evaluated the effect of R1R2 on SMC proliferation and migration in vitro. R1R2 peptide significantly inhibited A7r5 SMC growth after 48–96 hours in comparison with the scrambled peptide (1.5 ± 0.224 x 105, n = 5 vs. 2.740 ± 0.688 x 105, n = 5 at 48hr)(Fig. 6A). To determine whether R1R2 affects SMC migration, a wound healing assay was performed in SMCs stimulated with PDGF-BB. As shown in Fig. 6B, cells migrated more slowly in the presence of R1R2 than in the presence of scrambled peptide 30 hours post PDGF-stimulation. A quantitative analysis of SMC migration showed that R1R2 significantly reduced SMC migration in comparison with the scrambled peptide (130149 ± 53295, n = 3 vs. 313537 ± 66925, n = 3)(Fig. 6C). Together, these data show that R1R2 inhibits smooth muscle cell phenotype modulation, proliferation, and migration.


Collagen inhibitory peptide R1R2 mediates vascular remodeling by decreasing inflammation and smooth muscle cell activation.

Lee TH, Sottile J, Chiang HY - PLoS ONE (2015)

R1R2 reduces SMC proliferation and migration.(A) A7r5 SMC growth study over 96 hours. After serum starvation and treatment with R1R2 or scrambled peptides for 36 hours, cells were stimulated with 10% FBS over 96 hours. Cells were counted at the indicated time in the figure (n = 5). (B) A7r5 SMCs were pretreated with R1R2 or scramble peptide for 36 hours. SMCs underwent a scratch wound injury and were then stimulated with 10 ng/ml PDGF-BB for 30 hours. Representative images from the scratch wound assays are shown. (C) Quantitation of SMC migration in the scratch wound healing assay was performed by subtracting the cell-free area 30 hours after PDGF-BB stimulation from the cell-free area in the beginning (n = 5). * indicates p<0.05 and ** p< 0.01.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4326127&req=5

pone.0117356.g006: R1R2 reduces SMC proliferation and migration.(A) A7r5 SMC growth study over 96 hours. After serum starvation and treatment with R1R2 or scrambled peptides for 36 hours, cells were stimulated with 10% FBS over 96 hours. Cells were counted at the indicated time in the figure (n = 5). (B) A7r5 SMCs were pretreated with R1R2 or scramble peptide for 36 hours. SMCs underwent a scratch wound injury and were then stimulated with 10 ng/ml PDGF-BB for 30 hours. Representative images from the scratch wound assays are shown. (C) Quantitation of SMC migration in the scratch wound healing assay was performed by subtracting the cell-free area 30 hours after PDGF-BB stimulation from the cell-free area in the beginning (n = 5). * indicates p<0.05 and ** p< 0.01.
Mentions: SMC proliferation and migration contribute to intima and media thickening after arterial injury [44,45]. Therefore, we evaluated the effect of R1R2 on SMC proliferation and migration in vitro. R1R2 peptide significantly inhibited A7r5 SMC growth after 48–96 hours in comparison with the scrambled peptide (1.5 ± 0.224 x 105, n = 5 vs. 2.740 ± 0.688 x 105, n = 5 at 48hr)(Fig. 6A). To determine whether R1R2 affects SMC migration, a wound healing assay was performed in SMCs stimulated with PDGF-BB. As shown in Fig. 6B, cells migrated more slowly in the presence of R1R2 than in the presence of scrambled peptide 30 hours post PDGF-stimulation. A quantitative analysis of SMC migration showed that R1R2 significantly reduced SMC migration in comparison with the scrambled peptide (130149 ± 53295, n = 3 vs. 313537 ± 66925, n = 3)(Fig. 6C). Together, these data show that R1R2 inhibits smooth muscle cell phenotype modulation, proliferation, and migration.

Bottom Line: Morphometric analysis demonstrated that R1R2 reduced intima-media thickening compared to the control groups.This decrease was accompanied by decreased VCAM-1 and ICAM-1 expression.Our in vitro studies revealed that R1R2 attenuated SMC proliferation and migration, and also decreased monocyte adhesion and transendothelial migration through endothelial cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan.

ABSTRACT
The extracellular matrix (ECM) is a major constituent of the vessel wall. In addition to providing a structural scaffold, the ECM controls numerous cellular functions in both physiologic and pathologic settings. Vascular remodeling occurs after injury and is characterized by endothelial cell activation, inflammatory cell infiltration, phenotypic modulation of smooth muscle cells (SMCs), and augmented deposition of collagen-rich ECM. R1R2, a peptide derived from the bacterial adhesin SFS, with sequence homology to collagen, is known to inhibit collagen type I deposition in vitro by inhibiting the binding of fibronectin to collagen. However, the inhibitory effects of R1R2 during vascular remodeling have not been explored. We periadventitially delivered R1R2 to carotid arteries using pluronic gel in a vascular remodeling mouse model induced by blood flow cessation, and evaluated its effects on intima-media thickening, ECM deposition, SMC activation, and inflammatory cell infiltration. Morphometric analysis demonstrated that R1R2 reduced intima-media thickening compared to the control groups. R1R2 treatment also decreased collagen type I deposition in the vessel wall, and maintained SMC in the contractile phenotype. Interestingly, R1R2 dramatically reduced inflammatory cell infiltration into the vessel by ∼ 78%. This decrease was accompanied by decreased VCAM-1 and ICAM-1 expression. Our in vitro studies revealed that R1R2 attenuated SMC proliferation and migration, and also decreased monocyte adhesion and transendothelial migration through endothelial cells. Together, these data suggest that R1R2 attenuates vascular remodeling responses by decreasing inflammation and by modulating SMC proliferation and migration, and suggest that the R1R2 peptide may have therapeutic potential in treating occlusive vascular diseases.

Show MeSH
Related in: MedlinePlus