The Effects of Ganoderma lucidum Polysaccharides on the Proliferation of Cultured Human Aortic Smooth Muscle Cells and the Neointimal Hyperplasia of Mice
碩士 === 國立臺灣大學 === 解剖學暨生物細胞學研究所 === 98 === Vascular smooth muscle cell (VSMCs) proliferation, triggered by inflammatory response of the vascular wall, is considered to be the key event in the development of atherosclerosis and restenosis. Therefore, the identification of novel compounds with combined...
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ndltd-TW-098NTU053910112015-11-02T04:04:03Z http://ndltd.ncl.edu.tw/handle/46911078050653441747 The Effects of Ganoderma lucidum Polysaccharides on the Proliferation of Cultured Human Aortic Smooth Muscle Cells and the Neointimal Hyperplasia of Mice 探討靈芝多醣體對PDGF處理血管平滑肌細胞和小鼠血管內膜增生的影響 Wen-Yu Weng 翁郁雯 碩士 國立臺灣大學 解剖學暨生物細胞學研究所 98 Vascular smooth muscle cell (VSMCs) proliferation, triggered by inflammatory response of the vascular wall, is considered to be the key event in the development of atherosclerosis and restenosis. Therefore, the identification of novel compounds with combined anti-inflammatory and anti-proliferative properties may be an attractive therapeutic strategy for the prevention of cardiovascular diseases. A glucan-containing extract of Ganoderma lucidum-derived polysaccharides (EORP) has been proposed to possess immuno-modulatory functions and antitumor activities. However, its effects on the proliferation of VSMCs and the relative mechanisms remain unclear. In this study we aimed to examine the effects of EORP on the PDGF (platelet-derived growth factor)-treated human aortic smooth muscle cells (HASMCs) and neointimal hyperplasia in endothelia-denuded femoral artery of mice. EORP treatment inhibited proliferation of PDGF-treated HASMCs demonstrated by cell count, MTT (3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and 5-bromodeoxyuridine (BrdU) incorporation (MTT data, control:1; PDGF:1.38±0.01; PDGF+10 μg/mL EORP:1.03±0.02; cell count data, control:15.45±0.65×103; PDGF:23.25±0.05×103; BrdU data: control: 0.02±0.01; PDGF: 0.08±0.01; PDGF+10 μg/mL EORP: 0.02±0.00). Flow cytometry analysis showed EORP altered cell cycle distribution. EORP decreased cell proportion of S phase and increased cell proportion of G0/G1 phase (G0/G1 phase, contol:89.1±1.4 %; PDGF: 80.4±4.3 %, PDGF+10 μg/mL EORP: 85.7±3.8 %; S phase, control:1.7±0.9 %; PDGF: 5.8±2.8 %, PDGF+10 μg/mL EORP: 3.4±2.5 %). Western blot analysis demonstrated EORP downregulated PDGF-induced cyclin D1, cyclin E, CDK2 expression and upregulated p27 expression. Phosphorylation studies of MAPKs demonstrated that EORP suppressed PDGF-induced JNK/SAPK (stress-activated protein kinase) and p38 phosphorylation. For in vivo studies, oral EORP treatment reduces neointimal hyperplasia in endothelia-denuded femoral artery of mice (I/M ratio, endothelial denudation: 1.46±0.30; EORP+endothelial denudation: 0.67±0.03) and downregulated cell proliferation marker-PCNA (proliferating cell nuclear antigen) expression (PCNA positive cells ratio, endothelial denudation: 79.44±3.80 %; EORP+endothelial denudation: 60.78±2.65%). These results suggest that EORP may provide an effective novel approach to prevent vascular diseases through inhibition of vascular smooth muscle cells proliferation. Yuh-Lien Chen 陳玉怜 2010 學位論文 ; thesis 53 zh-TW |
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碩士 === 國立臺灣大學 === 解剖學暨生物細胞學研究所 === 98 === Vascular smooth muscle cell (VSMCs) proliferation, triggered by inflammatory response of the vascular wall, is considered to be the key event in the development of atherosclerosis and restenosis. Therefore, the identification of novel compounds with combined anti-inflammatory and anti-proliferative properties may be an attractive therapeutic strategy for the prevention of cardiovascular diseases. A glucan-containing extract of Ganoderma lucidum-derived polysaccharides (EORP) has been proposed to possess immuno-modulatory functions and antitumor activities. However, its effects on the proliferation of VSMCs and the relative mechanisms remain unclear. In this study we aimed to examine the effects of EORP on the PDGF (platelet-derived growth factor)-treated human aortic smooth muscle cells (HASMCs) and neointimal hyperplasia in endothelia-denuded femoral artery of mice. EORP treatment inhibited proliferation of PDGF-treated HASMCs demonstrated by cell count, MTT (3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and 5-bromodeoxyuridine (BrdU) incorporation (MTT data, control:1; PDGF:1.38±0.01; PDGF+10 μg/mL EORP:1.03±0.02; cell count data, control:15.45±0.65×103; PDGF:23.25±0.05×103; BrdU data: control: 0.02±0.01; PDGF: 0.08±0.01; PDGF+10 μg/mL EORP: 0.02±0.00). Flow cytometry analysis showed EORP altered cell cycle distribution. EORP decreased cell proportion of S phase and increased cell proportion of G0/G1 phase (G0/G1 phase, contol:89.1±1.4 %; PDGF: 80.4±4.3 %, PDGF+10 μg/mL EORP: 85.7±3.8 %; S phase, control:1.7±0.9 %; PDGF: 5.8±2.8 %, PDGF+10 μg/mL EORP: 3.4±2.5 %). Western blot analysis demonstrated EORP downregulated PDGF-induced cyclin D1, cyclin E, CDK2 expression and upregulated p27 expression. Phosphorylation studies of MAPKs demonstrated that EORP suppressed PDGF-induced JNK/SAPK (stress-activated protein kinase) and p38 phosphorylation. For in vivo studies, oral EORP treatment reduces neointimal hyperplasia in endothelia-denuded femoral artery of mice (I/M ratio, endothelial denudation: 1.46±0.30; EORP+endothelial denudation: 0.67±0.03) and downregulated cell proliferation marker-PCNA (proliferating cell nuclear antigen) expression (PCNA positive cells ratio, endothelial denudation: 79.44±3.80 %; EORP+endothelial denudation: 60.78±2.65%). These results suggest that EORP may provide an effective novel approach to prevent vascular diseases through inhibition of vascular smooth muscle cells proliferation.
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author2 |
Yuh-Lien Chen |
author_facet |
Yuh-Lien Chen Wen-Yu Weng 翁郁雯 |
author |
Wen-Yu Weng 翁郁雯 |
spellingShingle |
Wen-Yu Weng 翁郁雯 The Effects of Ganoderma lucidum Polysaccharides on the Proliferation of Cultured Human Aortic Smooth Muscle Cells and the Neointimal Hyperplasia of Mice |
author_sort |
Wen-Yu Weng |
title |
The Effects of Ganoderma lucidum Polysaccharides on the Proliferation of Cultured Human Aortic Smooth Muscle Cells and the Neointimal Hyperplasia of Mice |
title_short |
The Effects of Ganoderma lucidum Polysaccharides on the Proliferation of Cultured Human Aortic Smooth Muscle Cells and the Neointimal Hyperplasia of Mice |
title_full |
The Effects of Ganoderma lucidum Polysaccharides on the Proliferation of Cultured Human Aortic Smooth Muscle Cells and the Neointimal Hyperplasia of Mice |
title_fullStr |
The Effects of Ganoderma lucidum Polysaccharides on the Proliferation of Cultured Human Aortic Smooth Muscle Cells and the Neointimal Hyperplasia of Mice |
title_full_unstemmed |
The Effects of Ganoderma lucidum Polysaccharides on the Proliferation of Cultured Human Aortic Smooth Muscle Cells and the Neointimal Hyperplasia of Mice |
title_sort |
effects of ganoderma lucidum polysaccharides on the proliferation of cultured human aortic smooth muscle cells and the neointimal hyperplasia of mice |
publishDate |
2010 |
url |
http://ndltd.ncl.edu.tw/handle/46911078050653441747 |
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