Dieckol is a polyphenol compound isolated from brown algae that has anti-oxidant, anti-inflammatory, and anti-tumor activity. We examined the anti-angiogenic effects of dieckol in endothelial cells under hypoxic conditions. Treatment with $CoCl_2$, a hypoxic mimetic agent, increased proli...
Dieckol is a polyphenol compound isolated from brown algae that has anti-oxidant, anti-inflammatory, and anti-tumor activity. We examined the anti-angiogenic effects of dieckol in endothelial cells under hypoxic conditions. Treatment with $CoCl_2$, a hypoxic mimetic agent, increased proliferation, adhesion, migration, and tube formation in HUVECs, as well as vessel sprouting in rat aortic rings, which correlated well with increased expression of hypoxia-inducible factor 1-alpha ($HIF1{\alpha}$) and ${\beta}1$-integrin. Dieckol suppressed $CoCl_2$-induced adhesion, migration, and tube formation in HUVECs and vessel sprouting in rat aortic rings. Dieckol treatment decreased $CoCl_2$-induced overexpression of $HIF1{\alpha}$ and its downstream signaling molecules, including ${\beta}1$-integrin/Fak, Akt/eNOS, and p38 MAPK. These results suggest that dieckol is a novel angiogenesis inhibitor and a potential treatment for angiogenesis-dependent diseases in humans, such as malignant tumors.
Dieckol is a polyphenol compound isolated from brown algae that has anti-oxidant, anti-inflammatory, and anti-tumor activity. We examined the anti-angiogenic effects of dieckol in endothelial cells under hypoxic conditions. Treatment with $CoCl_2$, a hypoxic mimetic agent, increased proliferation, adhesion, migration, and tube formation in HUVECs, as well as vessel sprouting in rat aortic rings, which correlated well with increased expression of hypoxia-inducible factor 1-alpha ($HIF1{\alpha}$) and ${\beta}1$-integrin. Dieckol suppressed $CoCl_2$-induced adhesion, migration, and tube formation in HUVECs and vessel sprouting in rat aortic rings. Dieckol treatment decreased $CoCl_2$-induced overexpression of $HIF1{\alpha}$ and its downstream signaling molecules, including ${\beta}1$-integrin/Fak, Akt/eNOS, and p38 MAPK. These results suggest that dieckol is a novel angiogenesis inhibitor and a potential treatment for angiogenesis-dependent diseases in humans, such as malignant tumors.
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제안 방법
A, HUVECs were incubated in the absence or presence of 100 µM CoCl2 and 25 µM dieckol for 24 h and then harvested for western blotting; B, HUVECs in Matrigel were pretreated with a β1-integrin-neutralizing antibody (200 ng/mL) for 30 min before incubation with CoCl2 for 16 h, and tube formation was induced and then photographed.
Based on this result, we tested the effects of CoCl2 and dieckol on the expression of β1-integrin in endothelial cells.
Therefore, in this study we focused on the role of β1-integrin in hypoxia-induced angiogenesis in human umbilical vein endothelial cells (HUVECs), and particularly on the anti-angiogenic effects of dieckol, which was identified as an inhibitor of β1-integrin signaling in our recent screening study (Park and Jeon, 2012).
대상 데이터
Human umbilical vein endothelial cells (HUVECs) were obtained from the American Type Culture Collection (Manassas, VA, USA). Cells were cultured in M199 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 20% fetal bovine serum (FBS), 100 units/mL penicillin, 100 µg/mL streptomycin, 3 ng/mL basic fibroblast growth factor (Upstate Biotechnology, NY), and 5 units/mL heparin.
데이터처리
The significance of differences between the means were analyzed by Student’s ttest.
이론/모형
HUVEC migratory function, which is essential for angiogenesis, was examined using a modified Boyden chamber technique. A 24-well Transwell apparatus (Costar) was used, with each well containing a 6.
When the cells were 80–90% confluent, the culture medium was replaced with serum-free M199 for 12 h, after which the cultures were divided into 2 groups: with or without 25 µM dieckol. The viability of cells after dieckol treatment was assessed by the MTT [3-(4, 5-dimethylthiazol2-yl)-2,5-diphenyltetrazolium bromide] assay. HUVEC morphology was assessed with phase-contrast microscopy and cell counts were also calculated by hemocytometry.
성능/효과
Taken together, our results suggest that dieckol inhibits signaling downstream of HIF1α, including signaling mediated by β1-integrin, Fak, Akt/eNOS, and p38 MAPK, and these effects of dieckol were associated with inhibition of CoCl2-induced angiogenesis in HUVECs.
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