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Reduction of endoplasmic reticulum stress inhibits neointima formation after vascular injury.

Ishimura S, Furuhashi M, Mita T, Fuseya T, Watanabe Y, Hoshina K, Kokubu N, Inoue K, Yoshida H, Miura T - Sci Rep (2014)

Bottom Line: Furthermore, treatment with ER stress reducers, 4-phenylbutyrate (4-PBA) and tauroursodeoxycholic acid (TUDCA), decreased the intima-to-media ratio after wire injury by 50.0% and 72.8%, respectively.Chronic stimulation of CASMC with PDGF-BB activated the UPR, and treatment with 4-PBA and TUDCA significantly suppressed the PDGF-BB-induced ER stress markers in CASMC and the proliferation and migration of CASMC.Suppression of ER stress would be a novel strategy against post-angioplasty vascular restenosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, S-1, W-16, Chuo-ku, Sapporo 060-8543, Japan.

ABSTRACT
Endoplasmic reticulum (ER) stress and inappropriate adaptation through the unfolded protein response (UPR) are predominant features of pathological processes. However, little is known about the link between ER stress and endovascular injury. We investigated the involvement of ER stress in neointima hyperplasia after vascular injury. The femoral arteries of 7-8-week-old male mice were subjected to wire-induced vascular injury. After 4 weeks, immunohistological analysis showed that ER stress markers were upregulated in the hyperplastic neointima. Neointima formation was increased by 54.8% in X-box binding protein-1 (XBP1) heterozygous mice, a model of compromised UPR. Knockdown of Xbp1 in human coronary artery smooth muscle cells (CASMC) in vitro promoted cell proliferation and migration. Furthermore, treatment with ER stress reducers, 4-phenylbutyrate (4-PBA) and tauroursodeoxycholic acid (TUDCA), decreased the intima-to-media ratio after wire injury by 50.0% and 72.8%, respectively. Chronic stimulation of CASMC with PDGF-BB activated the UPR, and treatment with 4-PBA and TUDCA significantly suppressed the PDGF-BB-induced ER stress markers in CASMC and the proliferation and migration of CASMC. In conclusion, increased ER stress contributes to neointima formation after vascular injury, while UPR signaling downstream of XBP1 plays a suppressive role. Suppression of ER stress would be a novel strategy against post-angioplasty vascular restenosis.

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Neointima formation, cell proliferation, cell migration and inflammation in Xbp1 haploinsufficiency.(A), (B). Left femoral arteries of 7-8-week-old male Xbp1+/+ (n = 7) and Xbp1+/− (n = 7) mice were subjected to wire-induced vascular injury. After 4 weeks, EVG staining was performed in the left femoral artery (A). Scale bars: 50 µm. The extent of neointima formation was evaluated as intima-to-media ratio in the wire-injured artery of Xbp1+/+ and Xbp1+/− mice (B). *P < 0.05. (C), (D). Gene expression of unfolded protein response (UPR) markers in the IRE1α-XBP1 branch (C) and the other branches (D) in Xbp1-knockdown human coronary artery smooth muscle cells (CASMC). *P < 0.05. (E). Cell proliferation was assessed by an MTS assay in Xbp1-knockdown CASMC with 20 ng/ml platelet-derived growth factor-BB (PDGF-BB) for 48 h. *P < 0.05 vs. Control-PDGF-BB(-); †P < 0.05 vs. Xbp1 KD-PDGF-BB(-); ‡P < 0.05 vs. Control-PDGF-BB(+). (F). Cell migration was assessed by a scratch wound assay in Xbp1-knockdown CASMC treated with 20 ng/ml PDGF-BB for 8 h. Photographs were taken, and migration distance was measured by ImageJ. *P < 0.05. (G). Gene expression of PDGF receptors, PDGFR-α and PDGFR-β, in Xbp1-knockdown CASMC. *P < 0.05. (H). Western blotting analysis of ER stress markers in Xbp1-knockdown CASMC treated with 20 ng/ml PDGF-BB for 48 h. (I). Gene expression of inflammatory markers, IL-1β and IL-6, in Xbp1-knockdown CASMC treated with 20 ng/ml PDGF-BB for 48 h. *P < 0.05 vs. Control-PDGF-BB(−); †P < 0.05 vs. Xbp1 KD-PDGF-BB(−); ‡P < 0.05 vs. Control-PDGF-BB(+).
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f2: Neointima formation, cell proliferation, cell migration and inflammation in Xbp1 haploinsufficiency.(A), (B). Left femoral arteries of 7-8-week-old male Xbp1+/+ (n = 7) and Xbp1+/− (n = 7) mice were subjected to wire-induced vascular injury. After 4 weeks, EVG staining was performed in the left femoral artery (A). Scale bars: 50 µm. The extent of neointima formation was evaluated as intima-to-media ratio in the wire-injured artery of Xbp1+/+ and Xbp1+/− mice (B). *P < 0.05. (C), (D). Gene expression of unfolded protein response (UPR) markers in the IRE1α-XBP1 branch (C) and the other branches (D) in Xbp1-knockdown human coronary artery smooth muscle cells (CASMC). *P < 0.05. (E). Cell proliferation was assessed by an MTS assay in Xbp1-knockdown CASMC with 20 ng/ml platelet-derived growth factor-BB (PDGF-BB) for 48 h. *P < 0.05 vs. Control-PDGF-BB(-); †P < 0.05 vs. Xbp1 KD-PDGF-BB(-); ‡P < 0.05 vs. Control-PDGF-BB(+). (F). Cell migration was assessed by a scratch wound assay in Xbp1-knockdown CASMC treated with 20 ng/ml PDGF-BB for 8 h. Photographs were taken, and migration distance was measured by ImageJ. *P < 0.05. (G). Gene expression of PDGF receptors, PDGFR-α and PDGFR-β, in Xbp1-knockdown CASMC. *P < 0.05. (H). Western blotting analysis of ER stress markers in Xbp1-knockdown CASMC treated with 20 ng/ml PDGF-BB for 48 h. (I). Gene expression of inflammatory markers, IL-1β and IL-6, in Xbp1-knockdown CASMC treated with 20 ng/ml PDGF-BB for 48 h. *P < 0.05 vs. Control-PDGF-BB(−); †P < 0.05 vs. Xbp1 KD-PDGF-BB(−); ‡P < 0.05 vs. Control-PDGF-BB(+).

Mentions: To further investigate the association between vascular remodeling and ER stress in vivo, wire injury-mediated neointima hyperplasia was induced in XBP1 heterozygous (Xbp1+/−) mice as a model of compromised UPR. Thickness of the neointima after wire injury was significantly larger (by 54.8%) in the Xbp1+/− mice than in wild-type mice (Figure 2A, B).


Reduction of endoplasmic reticulum stress inhibits neointima formation after vascular injury.

Ishimura S, Furuhashi M, Mita T, Fuseya T, Watanabe Y, Hoshina K, Kokubu N, Inoue K, Yoshida H, Miura T - Sci Rep (2014)

Neointima formation, cell proliferation, cell migration and inflammation in Xbp1 haploinsufficiency.(A), (B). Left femoral arteries of 7-8-week-old male Xbp1+/+ (n = 7) and Xbp1+/− (n = 7) mice were subjected to wire-induced vascular injury. After 4 weeks, EVG staining was performed in the left femoral artery (A). Scale bars: 50 µm. The extent of neointima formation was evaluated as intima-to-media ratio in the wire-injured artery of Xbp1+/+ and Xbp1+/− mice (B). *P < 0.05. (C), (D). Gene expression of unfolded protein response (UPR) markers in the IRE1α-XBP1 branch (C) and the other branches (D) in Xbp1-knockdown human coronary artery smooth muscle cells (CASMC). *P < 0.05. (E). Cell proliferation was assessed by an MTS assay in Xbp1-knockdown CASMC with 20 ng/ml platelet-derived growth factor-BB (PDGF-BB) for 48 h. *P < 0.05 vs. Control-PDGF-BB(-); †P < 0.05 vs. Xbp1 KD-PDGF-BB(-); ‡P < 0.05 vs. Control-PDGF-BB(+). (F). Cell migration was assessed by a scratch wound assay in Xbp1-knockdown CASMC treated with 20 ng/ml PDGF-BB for 8 h. Photographs were taken, and migration distance was measured by ImageJ. *P < 0.05. (G). Gene expression of PDGF receptors, PDGFR-α and PDGFR-β, in Xbp1-knockdown CASMC. *P < 0.05. (H). Western blotting analysis of ER stress markers in Xbp1-knockdown CASMC treated with 20 ng/ml PDGF-BB for 48 h. (I). Gene expression of inflammatory markers, IL-1β and IL-6, in Xbp1-knockdown CASMC treated with 20 ng/ml PDGF-BB for 48 h. *P < 0.05 vs. Control-PDGF-BB(−); †P < 0.05 vs. Xbp1 KD-PDGF-BB(−); ‡P < 0.05 vs. Control-PDGF-BB(+).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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f2: Neointima formation, cell proliferation, cell migration and inflammation in Xbp1 haploinsufficiency.(A), (B). Left femoral arteries of 7-8-week-old male Xbp1+/+ (n = 7) and Xbp1+/− (n = 7) mice were subjected to wire-induced vascular injury. After 4 weeks, EVG staining was performed in the left femoral artery (A). Scale bars: 50 µm. The extent of neointima formation was evaluated as intima-to-media ratio in the wire-injured artery of Xbp1+/+ and Xbp1+/− mice (B). *P < 0.05. (C), (D). Gene expression of unfolded protein response (UPR) markers in the IRE1α-XBP1 branch (C) and the other branches (D) in Xbp1-knockdown human coronary artery smooth muscle cells (CASMC). *P < 0.05. (E). Cell proliferation was assessed by an MTS assay in Xbp1-knockdown CASMC with 20 ng/ml platelet-derived growth factor-BB (PDGF-BB) for 48 h. *P < 0.05 vs. Control-PDGF-BB(-); †P < 0.05 vs. Xbp1 KD-PDGF-BB(-); ‡P < 0.05 vs. Control-PDGF-BB(+). (F). Cell migration was assessed by a scratch wound assay in Xbp1-knockdown CASMC treated with 20 ng/ml PDGF-BB for 8 h. Photographs were taken, and migration distance was measured by ImageJ. *P < 0.05. (G). Gene expression of PDGF receptors, PDGFR-α and PDGFR-β, in Xbp1-knockdown CASMC. *P < 0.05. (H). Western blotting analysis of ER stress markers in Xbp1-knockdown CASMC treated with 20 ng/ml PDGF-BB for 48 h. (I). Gene expression of inflammatory markers, IL-1β and IL-6, in Xbp1-knockdown CASMC treated with 20 ng/ml PDGF-BB for 48 h. *P < 0.05 vs. Control-PDGF-BB(−); †P < 0.05 vs. Xbp1 KD-PDGF-BB(−); ‡P < 0.05 vs. Control-PDGF-BB(+).
Mentions: To further investigate the association between vascular remodeling and ER stress in vivo, wire injury-mediated neointima hyperplasia was induced in XBP1 heterozygous (Xbp1+/−) mice as a model of compromised UPR. Thickness of the neointima after wire injury was significantly larger (by 54.8%) in the Xbp1+/− mice than in wild-type mice (Figure 2A, B).

Bottom Line: Furthermore, treatment with ER stress reducers, 4-phenylbutyrate (4-PBA) and tauroursodeoxycholic acid (TUDCA), decreased the intima-to-media ratio after wire injury by 50.0% and 72.8%, respectively.Chronic stimulation of CASMC with PDGF-BB activated the UPR, and treatment with 4-PBA and TUDCA significantly suppressed the PDGF-BB-induced ER stress markers in CASMC and the proliferation and migration of CASMC.Suppression of ER stress would be a novel strategy against post-angioplasty vascular restenosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, S-1, W-16, Chuo-ku, Sapporo 060-8543, Japan.

ABSTRACT
Endoplasmic reticulum (ER) stress and inappropriate adaptation through the unfolded protein response (UPR) are predominant features of pathological processes. However, little is known about the link between ER stress and endovascular injury. We investigated the involvement of ER stress in neointima hyperplasia after vascular injury. The femoral arteries of 7-8-week-old male mice were subjected to wire-induced vascular injury. After 4 weeks, immunohistological analysis showed that ER stress markers were upregulated in the hyperplastic neointima. Neointima formation was increased by 54.8% in X-box binding protein-1 (XBP1) heterozygous mice, a model of compromised UPR. Knockdown of Xbp1 in human coronary artery smooth muscle cells (CASMC) in vitro promoted cell proliferation and migration. Furthermore, treatment with ER stress reducers, 4-phenylbutyrate (4-PBA) and tauroursodeoxycholic acid (TUDCA), decreased the intima-to-media ratio after wire injury by 50.0% and 72.8%, respectively. Chronic stimulation of CASMC with PDGF-BB activated the UPR, and treatment with 4-PBA and TUDCA significantly suppressed the PDGF-BB-induced ER stress markers in CASMC and the proliferation and migration of CASMC. In conclusion, increased ER stress contributes to neointima formation after vascular injury, while UPR signaling downstream of XBP1 plays a suppressive role. Suppression of ER stress would be a novel strategy against post-angioplasty vascular restenosis.

Show MeSH
Related in: MedlinePlus