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Potential contribution of phenotypically modulated smooth muscle cells and related inflammation in the development of experimental obstructive pulmonary vasculopathy in rats.

Otsuki S, Sawada H, Yodoya N, Shinohara T, Kato T, Ohashi H, Zhang E, Imanaka-Yoshida K, Shimpo H, Maruyama K, Komada Y, Mitani Y - PLoS ONE (2015)

Bottom Line: Immature SMC-rich intimal and plexiform lesions were proliferative and were infiltrated by macrophages, while fibrous intimal lesions were characterized by lower proliferative abilities and were infiltrated by few macrophages.Compared with controls, the number of perivascular macrophages was already higher at 3 weeks and progressively increased during the experimental period; gene expression of pulmonary hypertension-related inflammatory molecules, including IL6, MCP1, MMP9, cathepsin-S, and RANTES, was persistently or progressively up-regulated in lungs of experimental animals.We concluded that phenotypically modulated SMCs and related inflammation are potentially associated with the progression of experimental obstructive PVD.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Mie University Graduate School of Medicine, Tsu, Mie, Japan.

ABSTRACT
We tested the hypothesis that phenotypically modulated smooth muscle cells (SMCs) and related inflammation are associated with the progression of experimental occlusive pulmonary vascular disease (PVD). Occlusive PVD was induced by combined exposure to a vascular endothelial growth factor receptor tyrosine kinase inhibitor Sugen 5416 and hypobaric hypoxia for 3 weeks in rats, which were then returned to ambient air. Hemodynamic, morphometric, and immunohistochemical studies, as well as gene expression analyses, were performed at 3, 5, 8, and 13 weeks after the initial treatment (n = 78). Experimental animals developed pulmonary hypertension and right ventricular hypertrophy, and exhibited a progressive increase in indices of PVD, including cellular intimal thickening and intimal fibrosis. Cellular intimal lesions comprised α smooth muscle actin (α SMA)+, SM1+, SM2+/-, vimentin+ immature SMCs that were covered by endothelial monolayers, while fibrous intimal lesions typically included α SMA+, SM1+, SM2+, vimentin+/- mature SMCs. Plexiform lesions comprised α SMA+, vimentin+, SM1-, SM2- myofibroblasts covered by endothelial monolayers. Immature SMC-rich intimal and plexiform lesions were proliferative and were infiltrated by macrophages, while fibrous intimal lesions were characterized by lower proliferative abilities and were infiltrated by few macrophages. Compared with controls, the number of perivascular macrophages was already higher at 3 weeks and progressively increased during the experimental period; gene expression of pulmonary hypertension-related inflammatory molecules, including IL6, MCP1, MMP9, cathepsin-S, and RANTES, was persistently or progressively up-regulated in lungs of experimental animals. We concluded that phenotypically modulated SMCs and related inflammation are potentially associated with the progression of experimental obstructive PVD.

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Inflammatory cells in Sugen/hypoxia rats.A: Photomicrographs of CD68-positive macrophages and pulmonary vascular lesions in a Sugen/hypoxia rat 3 weeks after initial treatment. B: Number of perivascular CD68-positive macrophages per vessel in control, hypoxia, and Sugen/hypoxia groups at 3 and 5 weeks was compared with a one-way analysis of variance followed by Tukey-Kramer multiple comparison test. Number of perivascular CD68-positive macrophages per vessel (C) and the percentage of macrophage-positive intima (D) at different time points were compared with a one-way analysis of variance followed by Tukey-Kramer multiple comparison test. Values are mean ± SD. E: Positive correlation between the number of perivascular macrophages per vessel in a lung section and the percentage of occlusive lesions in a lung section (Pearson product-moment correlation coefficients). F: Percentage of the number of perivascular macrophages per vessel in that of total perivascular inflammatory cells, including macrophages, CD3+ T cells, and mast cells, at different time points were compared with a one-way analysis of variance followed by Tukey-Kramer multiple comparison test.
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pone.0118655.g007: Inflammatory cells in Sugen/hypoxia rats.A: Photomicrographs of CD68-positive macrophages and pulmonary vascular lesions in a Sugen/hypoxia rat 3 weeks after initial treatment. B: Number of perivascular CD68-positive macrophages per vessel in control, hypoxia, and Sugen/hypoxia groups at 3 and 5 weeks was compared with a one-way analysis of variance followed by Tukey-Kramer multiple comparison test. Number of perivascular CD68-positive macrophages per vessel (C) and the percentage of macrophage-positive intima (D) at different time points were compared with a one-way analysis of variance followed by Tukey-Kramer multiple comparison test. Values are mean ± SD. E: Positive correlation between the number of perivascular macrophages per vessel in a lung section and the percentage of occlusive lesions in a lung section (Pearson product-moment correlation coefficients). F: Percentage of the number of perivascular macrophages per vessel in that of total perivascular inflammatory cells, including macrophages, CD3+ T cells, and mast cells, at different time points were compared with a one-way analysis of variance followed by Tukey-Kramer multiple comparison test.

Mentions: Three and five weeks after initial treatment, perivascular macrophages were significantly increased in Sugen/hypoxia (0.51 ± 0.33, p = .0465 at 3 weeks; 0.51 ± 0.32, p = .0474 at 5 weeks) and in hypoxic rats (0.53 ± 0.28, p = .0342 at 3 weeks; 0.63 ± 0.27, p = .0045 at 5 weeks), compared with in control rats (0.20 ± 0.09 at 3 weeks; 0.20 ± 0.10 at 5 weeks) (Fig. 7A, B). In Sugen/hypoxia rats, the number of perivascular macrophages temporally increased until 13 weeks (2.6 ± 2.9, p = .0397 vs. 3 weeks) (Fig. 7C). Few macrophages infiltrated in the intima at 3 weeks. However, the proportion of macrophage-positive intima temporally increased until 13 weeks (22.6%, p = .0203 vs. 3 weeks) (Fig. 7D). The number of perivascular macrophages was positively correlated with the percentage of occlusive lesions (P<.0001, r = .7920) (Fig. 7E). Perivascular T cells or mast cells were not significantly increased in Sugen/hypoxia and in hypoxic rats at 3 or 5 weeks, compared with in control rats (S4 Fig.). However, in Sugen/hypoxia rats, the number of perivascular T cells (P = .0233 vs. 3 weeks) and mast cells (P<.0001 vs. 3, 5, and 8 weeks) temporally increased until 13 weeks (S4 Fig.). The percentage of the number of perivascular macrophages/ that of total perivascular inflammatory cells, including macrophages, CD3+ T cells, and mast cells, was 75.1% (22.6) at 3 weeks and was constant during the experimental period (Fig. 7F).


Potential contribution of phenotypically modulated smooth muscle cells and related inflammation in the development of experimental obstructive pulmonary vasculopathy in rats.

Otsuki S, Sawada H, Yodoya N, Shinohara T, Kato T, Ohashi H, Zhang E, Imanaka-Yoshida K, Shimpo H, Maruyama K, Komada Y, Mitani Y - PLoS ONE (2015)

Inflammatory cells in Sugen/hypoxia rats.A: Photomicrographs of CD68-positive macrophages and pulmonary vascular lesions in a Sugen/hypoxia rat 3 weeks after initial treatment. B: Number of perivascular CD68-positive macrophages per vessel in control, hypoxia, and Sugen/hypoxia groups at 3 and 5 weeks was compared with a one-way analysis of variance followed by Tukey-Kramer multiple comparison test. Number of perivascular CD68-positive macrophages per vessel (C) and the percentage of macrophage-positive intima (D) at different time points were compared with a one-way analysis of variance followed by Tukey-Kramer multiple comparison test. Values are mean ± SD. E: Positive correlation between the number of perivascular macrophages per vessel in a lung section and the percentage of occlusive lesions in a lung section (Pearson product-moment correlation coefficients). F: Percentage of the number of perivascular macrophages per vessel in that of total perivascular inflammatory cells, including macrophages, CD3+ T cells, and mast cells, at different time points were compared with a one-way analysis of variance followed by Tukey-Kramer multiple comparison test.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4340876&req=5

pone.0118655.g007: Inflammatory cells in Sugen/hypoxia rats.A: Photomicrographs of CD68-positive macrophages and pulmonary vascular lesions in a Sugen/hypoxia rat 3 weeks after initial treatment. B: Number of perivascular CD68-positive macrophages per vessel in control, hypoxia, and Sugen/hypoxia groups at 3 and 5 weeks was compared with a one-way analysis of variance followed by Tukey-Kramer multiple comparison test. Number of perivascular CD68-positive macrophages per vessel (C) and the percentage of macrophage-positive intima (D) at different time points were compared with a one-way analysis of variance followed by Tukey-Kramer multiple comparison test. Values are mean ± SD. E: Positive correlation between the number of perivascular macrophages per vessel in a lung section and the percentage of occlusive lesions in a lung section (Pearson product-moment correlation coefficients). F: Percentage of the number of perivascular macrophages per vessel in that of total perivascular inflammatory cells, including macrophages, CD3+ T cells, and mast cells, at different time points were compared with a one-way analysis of variance followed by Tukey-Kramer multiple comparison test.
Mentions: Three and five weeks after initial treatment, perivascular macrophages were significantly increased in Sugen/hypoxia (0.51 ± 0.33, p = .0465 at 3 weeks; 0.51 ± 0.32, p = .0474 at 5 weeks) and in hypoxic rats (0.53 ± 0.28, p = .0342 at 3 weeks; 0.63 ± 0.27, p = .0045 at 5 weeks), compared with in control rats (0.20 ± 0.09 at 3 weeks; 0.20 ± 0.10 at 5 weeks) (Fig. 7A, B). In Sugen/hypoxia rats, the number of perivascular macrophages temporally increased until 13 weeks (2.6 ± 2.9, p = .0397 vs. 3 weeks) (Fig. 7C). Few macrophages infiltrated in the intima at 3 weeks. However, the proportion of macrophage-positive intima temporally increased until 13 weeks (22.6%, p = .0203 vs. 3 weeks) (Fig. 7D). The number of perivascular macrophages was positively correlated with the percentage of occlusive lesions (P<.0001, r = .7920) (Fig. 7E). Perivascular T cells or mast cells were not significantly increased in Sugen/hypoxia and in hypoxic rats at 3 or 5 weeks, compared with in control rats (S4 Fig.). However, in Sugen/hypoxia rats, the number of perivascular T cells (P = .0233 vs. 3 weeks) and mast cells (P<.0001 vs. 3, 5, and 8 weeks) temporally increased until 13 weeks (S4 Fig.). The percentage of the number of perivascular macrophages/ that of total perivascular inflammatory cells, including macrophages, CD3+ T cells, and mast cells, was 75.1% (22.6) at 3 weeks and was constant during the experimental period (Fig. 7F).

Bottom Line: Immature SMC-rich intimal and plexiform lesions were proliferative and were infiltrated by macrophages, while fibrous intimal lesions were characterized by lower proliferative abilities and were infiltrated by few macrophages.Compared with controls, the number of perivascular macrophages was already higher at 3 weeks and progressively increased during the experimental period; gene expression of pulmonary hypertension-related inflammatory molecules, including IL6, MCP1, MMP9, cathepsin-S, and RANTES, was persistently or progressively up-regulated in lungs of experimental animals.We concluded that phenotypically modulated SMCs and related inflammation are potentially associated with the progression of experimental obstructive PVD.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Mie University Graduate School of Medicine, Tsu, Mie, Japan.

ABSTRACT
We tested the hypothesis that phenotypically modulated smooth muscle cells (SMCs) and related inflammation are associated with the progression of experimental occlusive pulmonary vascular disease (PVD). Occlusive PVD was induced by combined exposure to a vascular endothelial growth factor receptor tyrosine kinase inhibitor Sugen 5416 and hypobaric hypoxia for 3 weeks in rats, which were then returned to ambient air. Hemodynamic, morphometric, and immunohistochemical studies, as well as gene expression analyses, were performed at 3, 5, 8, and 13 weeks after the initial treatment (n = 78). Experimental animals developed pulmonary hypertension and right ventricular hypertrophy, and exhibited a progressive increase in indices of PVD, including cellular intimal thickening and intimal fibrosis. Cellular intimal lesions comprised α smooth muscle actin (α SMA)+, SM1+, SM2+/-, vimentin+ immature SMCs that were covered by endothelial monolayers, while fibrous intimal lesions typically included α SMA+, SM1+, SM2+, vimentin+/- mature SMCs. Plexiform lesions comprised α SMA+, vimentin+, SM1-, SM2- myofibroblasts covered by endothelial monolayers. Immature SMC-rich intimal and plexiform lesions were proliferative and were infiltrated by macrophages, while fibrous intimal lesions were characterized by lower proliferative abilities and were infiltrated by few macrophages. Compared with controls, the number of perivascular macrophages was already higher at 3 weeks and progressively increased during the experimental period; gene expression of pulmonary hypertension-related inflammatory molecules, including IL6, MCP1, MMP9, cathepsin-S, and RANTES, was persistently or progressively up-regulated in lungs of experimental animals. We concluded that phenotypically modulated SMCs and related inflammation are potentially associated with the progression of experimental obstructive PVD.

Show MeSH
Related in: MedlinePlus