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
Yi-Chung Chien, Ming-Jyh Sheu, Chieh-Hsi Wu, Wen-Hsin Lin, Ying-Yi Chen, Po-Liang Cheng, Hsu-Chen Cheng
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.
Copyright © 2012 Chien et al. This is an open access article distributed under the
Creative Commons Attribution License
1. Reagents and chemicals
3. Preparation of GLY extract
4. Cell culture
5. Cytotoxicity assay
6. Flow cytometric analysis
7. Wound healing assay
8. Cell migration assay
9. Determination of MMP-2 and MMP-9 by zymography
10. Western blotting analysis
11. Balloon angioplasty
12. Statistical analysis
1. GLY inhibited FBS-induced VSMCs proliferation
2. Effects of GLY on cell cycle
3. Non-cytotoxic doses of GLY inhibited FBS-stimulated VSMCs migration
4. Non-cytotoxic doses of GLY inhibits the Release of MMP-2 and MMP-9 in VSMCs Cells
5. Non-cytotoxic doses of GLY activated TIMPs, MMP-2, and MMP-9 expressions in VSMCs cells
6. Non-cytotoxic doses of GLY inhibited phosphorylated focal adhesion kinase and phosphorylated extracellular signal-regulated kinase
7. Non-cytotoxic doses of GLY inhibited the PI3K/Akt signaling in VSMCs cells
8. Extract of GLY inhibited balloon injury-induced neointima formation on the carotid artery
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