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. 2017 Jul 31;18(8):1664.
doi: 10.3390/ijms18081664.

Beneficial Effects of Galectin-3 Blockade in Vascular and Aortic Valve Alterations in an Experimental Pressure Overload Model

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Free PMC article

Beneficial Effects of Galectin-3 Blockade in Vascular and Aortic Valve Alterations in an Experimental Pressure Overload Model

Jaime Ibarrola et al. Int J Mol Sci. .
Free PMC article

Abstract

Galectin-3 (Gal-3) is involved in cardiovascular fibrosis and aortic valve (AV) calcification. We hypothesized that Gal-3 pharmacological inhibition with modified citrus pectin (MCP) could reduce aortic and AV remodeling in normotensive rats with pressure overload (PO). Six weeks after aortic constriction, vascular Gal-3 expression was up-regulated in male Wistar rats. Gal-3 overexpression was accompanied by an increase in the aortic media layer thickness, enhanced total collagen, and augmented expression of fibrotic mediators. Further, vascular inflammatory markers as well as inflammatory cells content were greater in aorta from PO rats. MCP treatment (100 mg/kg/day) prevented the increase in Gal-3, media thickness, fibrosis, and inflammation in the aorta of PO rats. Gal-3 levels were higher in AVs from PO rats. This paralleled enhanced AV fibrosis, inflammation, as well as greater expression of calcification markers. MCP treatment prevented the increase in Gal-3 as well as fibrosis, inflammation, and calcification in AVs. Overall, Gal-3 is overexpressed in aorta and AVs from PO rats. Gal-3 pharmacological inhibition blocks aortic and AV remodeling in experimental PO. Gal-3 could be a new therapeutic approach to delay the progression and the development of aortic remodeling and AV calcification in PO.

Keywords: Galectin-3; aorta; pressure overload; valve.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Pharmacological inhibition of Galectin-3 (Gal-3) decreases aortic wall thickness. Quantification of aortic Gal-3 mRNA levels in controls, pressure overload (PO) and PO + modified citrus pectin (MCP) rats (A); representative pictures of aortic sections immunostained for Gal-3 (B); haematoxylin/eosin staining of aortas (C); and quantification of aortic morphometry (D,E). Histogram bars represent the mean ± standard error of mean (SEM) of each group of rats in arbitrary units normalized to hypoxanthine phosphoribosyltransferase (HPRT) and β-actin for cDNA. Magnification 40×. (Control rats, n = 7; Pressure Overload rats, PO, n = 7; and Pressure Overload rats treated with MCP, PO + MCP, n = 7). * p < 0.05 vs. control group; $ p < 0.05 vs. PO group.
Figure 2
Figure 2
Pharmacological inhibition of Gal-3 prevents vascular fibrosis. Quantification of mRNA levels of Col1a1, fibronectin, α-smooth muscle actin (α-SMA), transforming growth factor (TGF)-β1, and connective tissue growth factor (CTGF) in aorta from controls, PO and PO + MCP rats (A); representative pictures of aortic sections immunostained for Sirius red, fibronectin, α-SMA, TGF-β1, and CTGF (B). Histogram bars represent the mean ± SEM of each group of rats in arbitrary units normalized to HPRT and β-actin for cDNA. Magnification 40×. (Control rats, n = 7; Pressure Overload rats, PO, n = 7; and Pressure Overload rats treated with MCP, PO + MCP, n = 7). * p < 0.05 vs. control group; $ p < 0.05 vs. PO group.
Figure 3
Figure 3
Pharmacological inhibition of Gal-3 blocks matrix metalloproteinase (MMP)-2 activity. Normalized values of the protein levels of MMP-2 (ng/mL), MMP-9 (pg/mL), and tissue inhibitor of metalloproteinase (TIMP)-2 (pg/mL) in aorta from controls, PO, and PO + MCP rats measured by ELISA (A); MMPs activities assessed by zymography (B); Representative pictures of aortic sections immunostained for MMP-2 (C). Histogram bars represent the mean ± SEM of each group of rats in arbitrary units. Magnification 40×. (Control rats, n = 7; Pressure Overload rats, PO, n = 7; and Pressure Overload rats treated with MCP, PO + MCP, n = 7). * p < 0.05 vs. control group; $ p < 0.05 vs. PO group.
Figure 4
Figure 4
Pharmacological inhibition of Gal-3 reduces vascular inflammation. Normalized values of the protein levels of interleukin (IL)-6 (pg/mL), IL-1β (pg/mL), tumor necrosis factor (TNF)-α (pg/mL), and monocyte chemoattractant protein (CCL)-2 (pg/mL) in aorta from controls, PO, and PO + MCP rats using ELISA (A); representative pictures of aortic sections immunostained for CCL-2, osteopontin, cd45, and cd68 (B). Histogram bars represent the mean ± SEM of each group of rats in arbitrary units. Magnification 40×. (Control rats, n = 7; Pressure Overload rats, PO, n = 7; and Pressure Overload rats treated with MCP, PO + MCP, n = 7). * p <0.05 vs. control group; $ p < 0.05 vs. PO group.
Figure 5
Figure 5
Pharmacological inhibition of Gal-3 diminishes aortic valve (AV) fibrosis. Immunostaining and quantification for Gal-3 in AVs from controls, PO, and PO + MCP rats (A). Haematoxylin/eosin staining of AVs and quantification of AV area (B); Sirius red staining and TGF-β1 immunostaining in AVs (C); MMP-2 and MMP-9 immunostaining in AVs (D). Histogram bars represent the mean ± SEM of each group of rats. Magnification 40×. (Control rats, n = 7; Pressure Overload rats, PO, n = 7; and Pressure Overload rats treated with MCP, PO + MCP, n = 7). * p < 0.05 vs. control group; $ p < 0.05 vs. PO group.
Figure 6
Figure 6
Pharmacological inhibition of Gal-3 reduces AV inflammation and calcification. Representative pictures of AV sections immunostained for cd45 and cd68 (A); Immunostaining (left panel) and quantification (right panel) for bone morphogenetic protein (BMP)-2, BMP-4, Runx2, and SRY-homeobOX-like (Sox-9, SRY: Sex-region-determining Y gene) in AVs from controls, PO, and PO + MCP rats (B). Arrows show the staining. Histogram bars represent the mean ± SEM of each group of rats. Magnification 40×. (Control rats, n = 7; Pressure Overload rats, PO, n = 7; and Pressure Overload rats treated with MCP, PO + MCP, n = 7). * p < 0.05 vs. control group; $ p < 0.05 vs. PO group.

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