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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

Molecular mechanisms of GLY on FBS-induced VSMCs. (A). Effects of GLY on TIMP-1, TIMP-2, MMP-2, MMP-9 proteins expression. VSMCs were treated with 0.125, 0.25 and 0.5 mg/mL for 24 h, and cell lysates were subjected to SDS-PAGE followed by Western blotting. (B). Dose-dependent effects of GLY on the protein expression level of FAK, the phosphorylated FAK, ERK1/2, the phosphorylated ERK1/2, PI3K, MMP-2, MMP-9, AKT and the phosphorylated AKT. VSMCs were treated with 0.125, 0.25 and 0.5 mg/mL of GLY for 24 h. The expression of these proteins were analyzed by Western blotting. β-actin was used as a loading control.
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Figure 4: Molecular mechanisms of GLY on FBS-induced VSMCs. (A). Effects of GLY on TIMP-1, TIMP-2, MMP-2, MMP-9 proteins expression. VSMCs were treated with 0.125, 0.25 and 0.5 mg/mL for 24 h, and cell lysates were subjected to SDS-PAGE followed by Western blotting. (B). Dose-dependent effects of GLY on the protein expression level of FAK, the phosphorylated FAK, ERK1/2, the phosphorylated ERK1/2, PI3K, MMP-2, MMP-9, AKT and the phosphorylated AKT. VSMCs were treated with 0.125, 0.25 and 0.5 mg/mL of GLY for 24 h. The expression of these proteins were analyzed by Western blotting. β-actin was used as a loading control.

Mentions: To further explore the modulation of pro-MMP and MMP activation by GLY, we determined TIMP-1/2 and MMP2/9 proteins expression levels. As shown in Figure4A, GLY strongly increased TIMP-1/2 activity and also decreased MMP-2/9 activity in a concentration-dependent manner.


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)

Molecular mechanisms of GLY on FBS-induced VSMCs. (A). Effects of GLY on TIMP-1, TIMP-2, MMP-2, MMP-9 proteins expression. VSMCs were treated with 0.125, 0.25 and 0.5 mg/mL for 24 h, and cell lysates were subjected to SDS-PAGE followed by Western blotting. (B). Dose-dependent effects of GLY on the protein expression level of FAK, the phosphorylated FAK, ERK1/2, the phosphorylated ERK1/2, PI3K, MMP-2, MMP-9, AKT and the phosphorylated AKT. VSMCs were treated with 0.125, 0.25 and 0.5 mg/mL of GLY for 24 h. The expression of these proteins were analyzed by Western blotting. β-actin was used as a loading control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Molecular mechanisms of GLY on FBS-induced VSMCs. (A). Effects of GLY on TIMP-1, TIMP-2, MMP-2, MMP-9 proteins expression. VSMCs were treated with 0.125, 0.25 and 0.5 mg/mL for 24 h, and cell lysates were subjected to SDS-PAGE followed by Western blotting. (B). Dose-dependent effects of GLY on the protein expression level of FAK, the phosphorylated FAK, ERK1/2, the phosphorylated ERK1/2, PI3K, MMP-2, MMP-9, AKT and the phosphorylated AKT. VSMCs were treated with 0.125, 0.25 and 0.5 mg/mL of GLY for 24 h. The expression of these proteins were analyzed by Western blotting. β-actin was used as a loading control.
Mentions: To further explore the modulation of pro-MMP and MMP activation by GLY, we determined TIMP-1/2 and MMP2/9 proteins expression levels. As shown in Figure4A, GLY strongly increased TIMP-1/2 activity and also decreased MMP-2/9 activity in a concentration-dependent manner.

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