Cyanidin 3-O-β-d-Glucoside from Black Bean Inhibits the Migration of Vascular Smooth Muscle Cells in the Presence of Adipocytes
Katsuya Yuui1★, Risa Kudo1, Shogo Kasuda1, Shigeru Ueshima2★ Corresponding author
- 1Department of Legal Medicine, Nara Medical University, Kashihara, Nara, JAPAN.
- 2Department of Food Science and Nutrition, Faculty of Agriculture, Kindai University, Nakamachi, Nara, JAPAN.
- 3T3-L1 cells within a range of 10–100 µM (Matsukawa et al., 2015). In the present study, we performed a preliminary cytotoxicity assessment to investigate the effects of low concentrations of components derived from natural products. Our findings revealed that a concentration of 20 µM does not exhibit cytotoxic effects, leading us to classify it as a physiological concentration. All controls utilized in this study were cultured in a medium supplemented with 0.1% methanol. Comprehensive supplementary methodologies were described as additional materials provided alongside the article (Supplemental material 1). SMCs obtained from the thoracic aorta of mice were prepared as previously described (Hou et al., 2004; Zhang et al., 2013). The SMCs utilized in this investigation were derived from cultured cells isolated from smooth muscle tissue obtained from the thoracic aorta of mice, in accordance with established protocols (Hou et al., 2004; Zhang et al., 2013). The established cultured cells were subsequently passaged for use in our previous report (Hou et al., 2007) and cultured at 37°C in a humidified atmosphere containing 5% CO₂. Oil Red O Staining Analyses The 3T3-L1 adipocytes were subjected to fixation with a 2.5% glutaraldehyde solution (lot number KWQ0074, Wako, Osaka, Japan) for a duration of 10 min. Following fixation, the cells were stained with Oil Red O for 15 min and subsequently rinsed with distilled water. Intracellular lipids were then extracted using isopropanol. The quantification of extracted lipids was then conducted utilizing a microplate.
CORRESPONDENCE
Katsuya Yuui
Department of Legal Medicine, Nara Medical University, Kashihara, Nara, JAPAN.
Received: 07-08-2024; Accepted: 16-12-2024.
Volume 21, Issue 4 · pp. 1459–1468 · PUBLISHED 2025 · DOI: 10.1177/09731296251313616
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ABSTRACT
Background: The potential of cyanidin 3-O-β-D-glucoside (C3G), known for its anti-obesity and anti-diabetic properties, in suppressing arteriosclerosis progression remains unexplored. Objectives: This study aimed to elucidate whether C3G prevents or ameliorates arteriosclerosis by investigating the effects of C3G on the transformation, proliferation, and migration of vascular smooth muscle cells (SMCs). Herein, we evaluated the effects of C3G on the migratory ability of SMCs in the presence of adipocytes. Materials and Methods: SMCs and 3T3-L1 adipocytes were cocultured in a Boyden chamber, with SMCs positioned in the upper chamber and pre-adipocytes in the lower chamber. Pre-adipocytes were initially seeded in the lower chamber and prompted to undergo differentiation over days 0, 2, 4, 6, and 8. SMCs were seeded in the upper chamber. The quantification of cells that migrated through the membrane was carried out. The migratory potential of SMCs within the SMC/adipocyte coculture (SACC) setup was assessed by placing the upper chamber into wells containing adipocytes, with or without 0.1% C3G. Results: The findings indicated that the accumulation of lipid droplets in adipocytes increased as the differentiation stage progressed. The amount of lipid droplets that accumulated upon the addition of C3G was significantly lower than that in the control. C3G was found to inhibit the active form of plasminogen activator inhibitor-1 (PAI-1) while enhancing the levels of the anti-inflammatory adipocytokine adiponectin. In the SACC, SMC migration increased alongside lipid droplet accumulation in adipocytes. We revealed that C3G decreased the active form of PAI-1 in the SACC and suppressed vascular SMC migration. Conclusion: C3G inhibited the migration of SMCs in the SACC, highlighting its potential as a compound that can prevent arteriosclerosis and thrombus formation progression by suppressing hyperplasia of the vascular intima.
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Yuui, K., Kudo, R., Kasuda, S., & Ueshima, S. (2025). Cyanidin 3-O-β-d-Glucoside from Black Bean Inhibits the Migration of Vascular Smooth Muscle Cells in the Presence of Adipocytes. Pharmacognosy Magazine, 21(4), 1459–1468. https://doi.org/10.1177/09731296251313616