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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.

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R1R2 decreases the binding between fibronectin (FN) and collagen.4 nM of FN was incubated with the indicated amount of R1R2 or scrambled peptide at 4°C overnight. The mixture of FN and R1R2 or scrambled peptide was added into collagen or BSA coated wells. Bound FN was detected with an anti-FN antibody followed by HRP-conjugated secondary antibody, and ABTS as described in Methods. The absorbance at 405nm was quantitated.
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pone.0117356.g001: R1R2 decreases the binding between fibronectin (FN) and collagen.4 nM of FN was incubated with the indicated amount of R1R2 or scrambled peptide at 4°C overnight. The mixture of FN and R1R2 or scrambled peptide was added into collagen or BSA coated wells. Bound FN was detected with an anti-FN antibody followed by HRP-conjugated secondary antibody, and ABTS as described in Methods. The absorbance at 405nm was quantitated.

Mentions: In order to assess whether inhibiting collagen type I deposition reduces vascular remodeling in an in vivo mouse model, we used the collagen inhibitory peptide R1R2 for this study. The R1R2 peptide is derived from two repeat domains of the bacterial adhesin SFS, and is highly homologous to collagen. R1R2 is known to inhibit collagen-FN binding, collagen type I deposition, and smooth muscle cell migration in vitro [29,30,33]. It is likely that R1R2 blocks collagen type I fibril formation around cells by binding to the collagen-binding domain of FN. To characterize the effects of our synthesized R1R2, and a newly generated scrambled peptide, we performed a solid-phase binding assay to determine the effect of R1R2 on FN binding to collagen. As shown in Fig. 1, R1R2 blocked the binding of FN to collagen type I in a dose-dependent manner. In contrast, the scrambled peptide had no effect on FN-collagen binding. Therefore, we used our synthesized R1R2 and scrambled peptides for further experiments in this study.


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 decreases the binding between fibronectin (FN) and collagen.4 nM of FN was incubated with the indicated amount of R1R2 or scrambled peptide at 4°C overnight. The mixture of FN and R1R2 or scrambled peptide was added into collagen or BSA coated wells. Bound FN was detected with an anti-FN antibody followed by HRP-conjugated secondary antibody, and ABTS as described in Methods. The absorbance at 405nm was quantitated.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0117356.g001: R1R2 decreases the binding between fibronectin (FN) and collagen.4 nM of FN was incubated with the indicated amount of R1R2 or scrambled peptide at 4°C overnight. The mixture of FN and R1R2 or scrambled peptide was added into collagen or BSA coated wells. Bound FN was detected with an anti-FN antibody followed by HRP-conjugated secondary antibody, and ABTS as described in Methods. The absorbance at 405nm was quantitated.
Mentions: In order to assess whether inhibiting collagen type I deposition reduces vascular remodeling in an in vivo mouse model, we used the collagen inhibitory peptide R1R2 for this study. The R1R2 peptide is derived from two repeat domains of the bacterial adhesin SFS, and is highly homologous to collagen. R1R2 is known to inhibit collagen-FN binding, collagen type I deposition, and smooth muscle cell migration in vitro [29,30,33]. It is likely that R1R2 blocks collagen type I fibril formation around cells by binding to the collagen-binding domain of FN. To characterize the effects of our synthesized R1R2, and a newly generated scrambled peptide, we performed a solid-phase binding assay to determine the effect of R1R2 on FN binding to collagen. As shown in Fig. 1, R1R2 blocked the binding of FN to collagen type I in a dose-dependent manner. In contrast, the scrambled peptide had no effect on FN-collagen binding. Therefore, we used our synthesized R1R2 and scrambled peptides for further experiments in this study.

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