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A Chinese herbal formula "Gan-Lu-Yin" suppresses vascular smooth muscle cell migration by inhibiting matrix metalloproteinase-2/9 through the PI3K/AKT and ERK signaling pathways.

Chien YC, Sheu MJ, Wu CH, Lin WH, Chen YY, Cheng PL, Cheng HC - BMC Complement Altern Med (2012)

Bottom Line: Our results showed that GLY significantly decreased the thickness of neointima.The inhibition by non-cytoxic doses of GLY of VSMCs migration was through its negative regulatory effects on phosphorylated ERK1/2, PI3K/AKT, and FAK.These observations provide a mechanism of GLY in attenuating cell migration, thus as a potential intervention for restenosis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Life Sciences, and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.

ABSTRACT

Background: This study was to explore the effects of Gan-Lu-Yin (GLY) on the migration of vascular smooth muscle cells (VSMCs) induced by fetal bovine serum and on neointima formation in a rat model of carotid artery balloon injury.

Methods: VSMCs were treated with different concentrations of GLY, and then analyzed with Flow cytometric analysis, zymography, transwell, and western blotting. SD rats received balloon-injury were analyzed with H&E staining.

Results: Our results showed that GLY significantly decreased the thickness of neointima. The inhibition by non-cytoxic doses of GLY of VSMCs migration was through its negative regulatory effects on phosphorylated ERK1/2, PI3K/AKT, and FAK. The data showed that GLY can inhibit the migration of VSMCs cells, and might block injury-induced neointima hyperplasia via the inhibition of VSMCs migration, without inducing apoptosis.

Conclusions: These observations provide a mechanism of GLY in attenuating cell migration, thus as a potential intervention for restenosis.

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Related in: MedlinePlus

Effects of GLY on cell viability and cell cycle analysis. (A). Effects of GLY on cell growth of VSMCs by MTT assay. The cells were incubated for 24 h with 10% fetal bovine serum alone (control) or with different concentrations of GLY (0.0625, 0.125, 0.25, 0.5, 1 and 2 mg/mL). (B). Flow cytometric analysis of GLY on the cell cycle of VSMCs cells. All the cells were treated with 15% fetal bovine serum with the addition of GLY at 0.0625,0.125,0.25,0.5,1 mg/mL for 18 h. (C). The percentage of subG1 under flow cytometric analysis. Values are means of three separate experiments, with standard errors represented by vertical bars. Mean value was significantly different from that of the control group :* p < 0.05,** p < 0.01, *** p < 0.001.
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Figure 1: Effects of GLY on cell viability and cell cycle analysis. (A). Effects of GLY on cell growth of VSMCs by MTT assay. The cells were incubated for 24 h with 10% fetal bovine serum alone (control) or with different concentrations of GLY (0.0625, 0.125, 0.25, 0.5, 1 and 2 mg/mL). (B). Flow cytometric analysis of GLY on the cell cycle of VSMCs cells. All the cells were treated with 15% fetal bovine serum with the addition of GLY at 0.0625,0.125,0.25,0.5,1 mg/mL for 18 h. (C). The percentage of subG1 under flow cytometric analysis. Values are means of three separate experiments, with standard errors represented by vertical bars. Mean value was significantly different from that of the control group :* p < 0.05,** p < 0.01, *** p < 0.001.

Mentions: Since outgrowth of VSMCs has been regarded as the major factor leading to restenosis, we performed the MTT assay to determine the inhibitory effects of GLY on cell viability of VSMCs. As shown in Figure1A, GLY inhibited VSMCs viability in a concentration-dependent manner. The inhibitory effect of GLY on cell viability became significant at 0.5 mg/mL (P < 0.05), 1 mg/mL (P < 0.01) and 2 mg/mL (P < 0.001) after 24 h incubation.


A Chinese herbal formula "Gan-Lu-Yin" suppresses vascular smooth muscle cell migration by inhibiting matrix metalloproteinase-2/9 through the PI3K/AKT and ERK signaling pathways.

Chien YC, Sheu MJ, Wu CH, Lin WH, Chen YY, Cheng PL, Cheng HC - BMC Complement Altern Med (2012)

Effects of GLY on cell viability and cell cycle analysis. (A). Effects of GLY on cell growth of VSMCs by MTT assay. The cells were incubated for 24 h with 10% fetal bovine serum alone (control) or with different concentrations of GLY (0.0625, 0.125, 0.25, 0.5, 1 and 2 mg/mL). (B). Flow cytometric analysis of GLY on the cell cycle of VSMCs cells. All the cells were treated with 15% fetal bovine serum with the addition of GLY at 0.0625,0.125,0.25,0.5,1 mg/mL for 18 h. (C). The percentage of subG1 under flow cytometric analysis. Values are means of three separate experiments, with standard errors represented by vertical bars. Mean value was significantly different from that of the control group :* p < 0.05,** p < 0.01, *** p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3475136&req=5

Figure 1: Effects of GLY on cell viability and cell cycle analysis. (A). Effects of GLY on cell growth of VSMCs by MTT assay. The cells were incubated for 24 h with 10% fetal bovine serum alone (control) or with different concentrations of GLY (0.0625, 0.125, 0.25, 0.5, 1 and 2 mg/mL). (B). Flow cytometric analysis of GLY on the cell cycle of VSMCs cells. All the cells were treated with 15% fetal bovine serum with the addition of GLY at 0.0625,0.125,0.25,0.5,1 mg/mL for 18 h. (C). The percentage of subG1 under flow cytometric analysis. Values are means of three separate experiments, with standard errors represented by vertical bars. Mean value was significantly different from that of the control group :* p < 0.05,** p < 0.01, *** p < 0.001.
Mentions: Since outgrowth of VSMCs has been regarded as the major factor leading to restenosis, we performed the MTT assay to determine the inhibitory effects of GLY on cell viability of VSMCs. As shown in Figure1A, GLY inhibited VSMCs viability in a concentration-dependent manner. The inhibitory effect of GLY on cell viability became significant at 0.5 mg/mL (P < 0.05), 1 mg/mL (P < 0.01) and 2 mg/mL (P < 0.001) after 24 h incubation.

Bottom Line: Our results showed that GLY significantly decreased the thickness of neointima.The inhibition by non-cytoxic doses of GLY of VSMCs migration was through its negative regulatory effects on phosphorylated ERK1/2, PI3K/AKT, and FAK.These observations provide a mechanism of GLY in attenuating cell migration, thus as a potential intervention for restenosis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Life Sciences, and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan.

ABSTRACT

Background: This study was to explore the effects of Gan-Lu-Yin (GLY) on the migration of vascular smooth muscle cells (VSMCs) induced by fetal bovine serum and on neointima formation in a rat model of carotid artery balloon injury.

Methods: VSMCs were treated with different concentrations of GLY, and then analyzed with Flow cytometric analysis, zymography, transwell, and western blotting. SD rats received balloon-injury were analyzed with H&E staining.

Results: Our results showed that GLY significantly decreased the thickness of neointima. The inhibition by non-cytoxic doses of GLY of VSMCs migration was through its negative regulatory effects on phosphorylated ERK1/2, PI3K/AKT, and FAK. The data showed that GLY can inhibit the migration of VSMCs cells, and might block injury-induced neointima hyperplasia via the inhibition of VSMCs migration, without inducing apoptosis.

Conclusions: These observations provide a mechanism of GLY in attenuating cell migration, thus as a potential intervention for restenosis.

Show MeSH
Related in: MedlinePlus