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Murrayafoline A Induces a G0/G1-Phase Arrest in Platelet-Derived Growth Factor-Stimulated Vascular Smooth Muscle Cells.

Han JH, Kim Y, Jung SH, Lee JJ, Park HS, Song GY, Cuong NM, Kim YH, Myung CS - Korean J. Physiol. Pharmacol. (2015)

Bottom Line: Abnormally high activity of platelet-derived growth factor (PDGF) is believed to play a central role in the etiology of these pathophysiological situations.Here, we investigated the anti-proliferative effects and possible mechanism(s) of murrayafoline A, a carbazole alkaloid isolated from Glycosmis stenocarpa Guillamin (Rutaceae), on PDGF-BB-stimulated VSMCs.These results indicate that murrayafoline A may be useful in preventing the progression of vascular complications such as restenosis after percutaneous transluminal coronary angioplasty and atherosclerosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Chungnam National University College of Pharmacy, Daejeon 305-764, Korea.

ABSTRACT
The increased potential for vascular smooth muscle cell (VSMC) growth is a key abnormality in the development of atherosclerosis and post-angioplasty restenosis. Abnormally high activity of platelet-derived growth factor (PDGF) is believed to play a central role in the etiology of these pathophysiological situations. Here, we investigated the anti-proliferative effects and possible mechanism(s) of murrayafoline A, a carbazole alkaloid isolated from Glycosmis stenocarpa Guillamin (Rutaceae), on PDGF-BB-stimulated VSMCs. Murrayafoline A inhibited the PDGF-BB-stimulated proliferation of VSMCs in a concentration-dependent manner, as measured using a non-radioactive colorimetric WST-1 assay and direct cell counting. Furthermore, murrayafoline A suppressed the PDGF-BB-stimulated progression through G0/G1 to S phase of the cell cycle, as measured by [(3)H]-thymidine incorporation assay and cell cycle progression analysis. This anti-proliferative action of murrayafoline A, arresting cell cycle progression at G0/G1 phase in PDGF-BB-stimulated VSMCs, was mediated via down-regulation of the expression of cyclin D1, cyclin E, cyclin-dependent kinase (CDK)2, CDK4, and proliferating cell nuclear antigen (PCNA), and the phosphorylation of retinoblastoma protein (pRb). These results indicate that murrayafoline A may be useful in preventing the progression of vascular complications such as restenosis after percutaneous transluminal coronary angioplasty and atherosclerosis.

No MeSH data available.


Related in: MedlinePlus

Effects of murrayafoline A on VSMC proliferation and viability. VSMCs cultured in serum-free medium were stimulated with 50 ng/ml PDGF-BB for 24 h and the effects of various concentrations of murrayafoline A (1-5 µM) on cell proliferation and viability were measured as described in the Experimental Section. (A) Optical densities at 450 nm, as determined in the WST-1 assay (n=3). (B) Cell numbers counted using a hemocytometer (n=4). (C) Viability as determined by the WST-1 assay (n=4) for various incubation times (0-48 h). All values are expressed as means±SEM. Statistical differences from the PDGF-BB control (PDGF-BB-stimulated, but no murrayafoline A) are indicated by *p<0.05 and **p<0.01.
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Figure 1: Effects of murrayafoline A on VSMC proliferation and viability. VSMCs cultured in serum-free medium were stimulated with 50 ng/ml PDGF-BB for 24 h and the effects of various concentrations of murrayafoline A (1-5 µM) on cell proliferation and viability were measured as described in the Experimental Section. (A) Optical densities at 450 nm, as determined in the WST-1 assay (n=3). (B) Cell numbers counted using a hemocytometer (n=4). (C) Viability as determined by the WST-1 assay (n=4) for various incubation times (0-48 h). All values are expressed as means±SEM. Statistical differences from the PDGF-BB control (PDGF-BB-stimulated, but no murrayafoline A) are indicated by *p<0.05 and **p<0.01.

Mentions: To investigate whether murrayafoline A could inhibit VSMC proliferation, a non-radioactive colorimetric WST-1 assay and direct cell counting were performed. PDGF-BB (50 ng/ml) increased VSMC proliferation by about two fold compared with unstimulated VSMCs (Fig. 1A). Murrayafoline A decreased PDGF-BB-stimulated VSMC proliferation in a concentration-dependent manner. Fig. 1B shows that the cell number decreased significantly, to 4.5±1.1×104, 2.9±0.8×104, and 2.2±0.1×104 cells/well, as the concentration of murrayafoline A was increased to 1, 3, and 5 µM, respectively. The number of cells increased significantly after 50 ng/ml PDGF-BB treatment (6.5±1.5×104 cells/well) compared with the unstimulated group (2.6±0.8×104 cells/well). Treatment with the highest concentration of murrayafoline A (5 µM) for various incubation times (6-48 h) showed no VSMC cytotoxicity in serum-free medium (Fig. 1C), indicating that the anti-proliferative effect of murrayafoline A on VSMCs was not due simply to a cytotoxic effect.


Murrayafoline A Induces a G0/G1-Phase Arrest in Platelet-Derived Growth Factor-Stimulated Vascular Smooth Muscle Cells.

Han JH, Kim Y, Jung SH, Lee JJ, Park HS, Song GY, Cuong NM, Kim YH, Myung CS - Korean J. Physiol. Pharmacol. (2015)

Effects of murrayafoline A on VSMC proliferation and viability. VSMCs cultured in serum-free medium were stimulated with 50 ng/ml PDGF-BB for 24 h and the effects of various concentrations of murrayafoline A (1-5 µM) on cell proliferation and viability were measured as described in the Experimental Section. (A) Optical densities at 450 nm, as determined in the WST-1 assay (n=3). (B) Cell numbers counted using a hemocytometer (n=4). (C) Viability as determined by the WST-1 assay (n=4) for various incubation times (0-48 h). All values are expressed as means±SEM. Statistical differences from the PDGF-BB control (PDGF-BB-stimulated, but no murrayafoline A) are indicated by *p<0.05 and **p<0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4553401&req=5

Figure 1: Effects of murrayafoline A on VSMC proliferation and viability. VSMCs cultured in serum-free medium were stimulated with 50 ng/ml PDGF-BB for 24 h and the effects of various concentrations of murrayafoline A (1-5 µM) on cell proliferation and viability were measured as described in the Experimental Section. (A) Optical densities at 450 nm, as determined in the WST-1 assay (n=3). (B) Cell numbers counted using a hemocytometer (n=4). (C) Viability as determined by the WST-1 assay (n=4) for various incubation times (0-48 h). All values are expressed as means±SEM. Statistical differences from the PDGF-BB control (PDGF-BB-stimulated, but no murrayafoline A) are indicated by *p<0.05 and **p<0.01.
Mentions: To investigate whether murrayafoline A could inhibit VSMC proliferation, a non-radioactive colorimetric WST-1 assay and direct cell counting were performed. PDGF-BB (50 ng/ml) increased VSMC proliferation by about two fold compared with unstimulated VSMCs (Fig. 1A). Murrayafoline A decreased PDGF-BB-stimulated VSMC proliferation in a concentration-dependent manner. Fig. 1B shows that the cell number decreased significantly, to 4.5±1.1×104, 2.9±0.8×104, and 2.2±0.1×104 cells/well, as the concentration of murrayafoline A was increased to 1, 3, and 5 µM, respectively. The number of cells increased significantly after 50 ng/ml PDGF-BB treatment (6.5±1.5×104 cells/well) compared with the unstimulated group (2.6±0.8×104 cells/well). Treatment with the highest concentration of murrayafoline A (5 µM) for various incubation times (6-48 h) showed no VSMC cytotoxicity in serum-free medium (Fig. 1C), indicating that the anti-proliferative effect of murrayafoline A on VSMCs was not due simply to a cytotoxic effect.

Bottom Line: Abnormally high activity of platelet-derived growth factor (PDGF) is believed to play a central role in the etiology of these pathophysiological situations.Here, we investigated the anti-proliferative effects and possible mechanism(s) of murrayafoline A, a carbazole alkaloid isolated from Glycosmis stenocarpa Guillamin (Rutaceae), on PDGF-BB-stimulated VSMCs.These results indicate that murrayafoline A may be useful in preventing the progression of vascular complications such as restenosis after percutaneous transluminal coronary angioplasty and atherosclerosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Chungnam National University College of Pharmacy, Daejeon 305-764, Korea.

ABSTRACT
The increased potential for vascular smooth muscle cell (VSMC) growth is a key abnormality in the development of atherosclerosis and post-angioplasty restenosis. Abnormally high activity of platelet-derived growth factor (PDGF) is believed to play a central role in the etiology of these pathophysiological situations. Here, we investigated the anti-proliferative effects and possible mechanism(s) of murrayafoline A, a carbazole alkaloid isolated from Glycosmis stenocarpa Guillamin (Rutaceae), on PDGF-BB-stimulated VSMCs. Murrayafoline A inhibited the PDGF-BB-stimulated proliferation of VSMCs in a concentration-dependent manner, as measured using a non-radioactive colorimetric WST-1 assay and direct cell counting. Furthermore, murrayafoline A suppressed the PDGF-BB-stimulated progression through G0/G1 to S phase of the cell cycle, as measured by [(3)H]-thymidine incorporation assay and cell cycle progression analysis. This anti-proliferative action of murrayafoline A, arresting cell cycle progression at G0/G1 phase in PDGF-BB-stimulated VSMCs, was mediated via down-regulation of the expression of cyclin D1, cyclin E, cyclin-dependent kinase (CDK)2, CDK4, and proliferating cell nuclear antigen (PCNA), and the phosphorylation of retinoblastoma protein (pRb). These results indicate that murrayafoline A may be useful in preventing the progression of vascular complications such as restenosis after percutaneous transluminal coronary angioplasty and atherosclerosis.

No MeSH data available.


Related in: MedlinePlus