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Anti-angiogenic effect of Nelumbo nucifera leaf extracts in human umbilical vein endothelial cells with antioxidant potential.

Lee JS, Shukla S, Kim JA, Kim M - PLoS ONE (2015)

Bottom Line: In this study, N. nucifera leaf extracts displayed potent antioxidant and inhibitory effects on VEGF-induced angiogenesis.N. nucifera exerted an inhibitory effect on VEGF-induced proliferation and tube formation, as well as CAM angiogenesis in vivo.Moreover, N. nucifera leaf extracts significantly blocked VEGF-induced ROS production in HUVECs, confirming their possible anti-angiogenic mechanism.

View Article: PubMed Central - PubMed

Affiliation: Department of Food Science and Technology, Yeungnam University, Gyeongsan, Gyeongbuk, 712-749, Republic of Korea.

ABSTRACT
Nelumbo nucifera Gaertn (Nymphaeaceae) has long been used as a traditional herb in Chinese, Japanese, Indian, and Korean medicinal practices since prehistoric times and flourishes today as the primary form of medicine. This study reports for the first time the potent ability of N. nucifera leaf extracts to inhibit vascular endothelial growth factor (VEGF)-induced angiogenesis in vitro and in vivo, as well as their antioxidant efficacy in various scavenging models and an analysis of their chemical composition. In vivo anti-angiogenic activity was evaluated in a chick chorioallantoic membrane (CAM) model using fertilized chicken eggs, in human umbilical vein endothelial cells (HUVECs) by using cell viability, cell proliferation and tube formation assays, and by determining intracellular reactive oxygen species (ROS) in vitro. The antioxidant efficacy of N. nucifera leaf extracts was determined in various scavenging models, including total phenolic and flavonoid content. The chemical composition of N. nucifera leaf extracts was determined by GC-MS analysis, which revealed the presence of different phytochemicals. The IC50 values for the DPPH radical scavenging activities of water and methanol extracts were found to be 1699.47 and 514.36 μg ml(-1), and their total phenolic and flavonoid contents were 85.01 ± 2.32 and 147.63 ± 2.23 mg GAE g dry mass(-1) and 35.38 ± 1.32 and 41.86 ± 1.07 mg QA g dry mass(-1), respectively. N. nucifera leaf extracts (10-100 μg ml(-1)) exhibited significant dose-dependent inhibition of VEGF-induced angiogenesis, as well as VEGF-induced proliferation and tube formation in HUVECs. In this study, N. nucifera leaf extracts displayed potent antioxidant and inhibitory effects on VEGF-induced angiogenesis. N. nucifera exerted an inhibitory effect on VEGF-induced proliferation and tube formation, as well as CAM angiogenesis in vivo. Moreover, N. nucifera leaf extracts significantly blocked VEGF-induced ROS production in HUVECs, confirming their possible anti-angiogenic mechanism.

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Inhibitory effects of water and methanol extracts of N. nucifera leaves on VEGF-induced angiogenesis.(a) The CAM of a 10-d-old chick embryo was separately exposed to PBS (control) and VEGF (20 ng ml−1) by means of filter disks. After 30 min, extracts were introduced on top of the CAMs. After 72 h of incubation, the CAM tissue directly beneath each filter disk was resected, and digital images of the CAM sections were captured. (b) The bar graph represents the number of new branches formed from existing blood vessels. Photographs were imported into an image software program to visualize the new vessel branch points. Data are shown as the mean ± SD. *: p<0.05 compared with untreated control; #: p<0.05 compared with VEGF-treated CAM samples. None of the tested concentrations (10, 50, and 100 μg ml−1) of the water as well as the methanol extract display statistically significant differences with respect to each other.
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pone.0118552.g003: Inhibitory effects of water and methanol extracts of N. nucifera leaves on VEGF-induced angiogenesis.(a) The CAM of a 10-d-old chick embryo was separately exposed to PBS (control) and VEGF (20 ng ml−1) by means of filter disks. After 30 min, extracts were introduced on top of the CAMs. After 72 h of incubation, the CAM tissue directly beneath each filter disk was resected, and digital images of the CAM sections were captured. (b) The bar graph represents the number of new branches formed from existing blood vessels. Photographs were imported into an image software program to visualize the new vessel branch points. Data are shown as the mean ± SD. *: p<0.05 compared with untreated control; #: p<0.05 compared with VEGF-treated CAM samples. None of the tested concentrations (10, 50, and 100 μg ml−1) of the water as well as the methanol extract display statistically significant differences with respect to each other.

Mentions: The ability of the water and methanol extracts of N. nucifera leaves to inhibit angiogenesis in vivo was determined using the CAM assay. The numbers of blood vessel branch points significantly increased upon VEGF treatment (Fig. 3), compared with the PBS-treated control group (Fig. 3). However, treatment with both the water and methanol leaf extracts of N. nucifera significantly suppressed VEGF-induced angiogenesis in a dose-dependent manner (Fig. 3). In strong support of these results, it was previously found that alliin from garlic displayed inhibitory activity towards fibroblast growth factor-2-induced tube formation and in vivo CAM angiogenesis [29]. Alliin was also found to inhibit VEGF-induced angiogenesis in the CAM [29].


Anti-angiogenic effect of Nelumbo nucifera leaf extracts in human umbilical vein endothelial cells with antioxidant potential.

Lee JS, Shukla S, Kim JA, Kim M - PLoS ONE (2015)

Inhibitory effects of water and methanol extracts of N. nucifera leaves on VEGF-induced angiogenesis.(a) The CAM of a 10-d-old chick embryo was separately exposed to PBS (control) and VEGF (20 ng ml−1) by means of filter disks. After 30 min, extracts were introduced on top of the CAMs. After 72 h of incubation, the CAM tissue directly beneath each filter disk was resected, and digital images of the CAM sections were captured. (b) The bar graph represents the number of new branches formed from existing blood vessels. Photographs were imported into an image software program to visualize the new vessel branch points. Data are shown as the mean ± SD. *: p<0.05 compared with untreated control; #: p<0.05 compared with VEGF-treated CAM samples. None of the tested concentrations (10, 50, and 100 μg ml−1) of the water as well as the methanol extract display statistically significant differences with respect to each other.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4340789&req=5

pone.0118552.g003: Inhibitory effects of water and methanol extracts of N. nucifera leaves on VEGF-induced angiogenesis.(a) The CAM of a 10-d-old chick embryo was separately exposed to PBS (control) and VEGF (20 ng ml−1) by means of filter disks. After 30 min, extracts were introduced on top of the CAMs. After 72 h of incubation, the CAM tissue directly beneath each filter disk was resected, and digital images of the CAM sections were captured. (b) The bar graph represents the number of new branches formed from existing blood vessels. Photographs were imported into an image software program to visualize the new vessel branch points. Data are shown as the mean ± SD. *: p<0.05 compared with untreated control; #: p<0.05 compared with VEGF-treated CAM samples. None of the tested concentrations (10, 50, and 100 μg ml−1) of the water as well as the methanol extract display statistically significant differences with respect to each other.
Mentions: The ability of the water and methanol extracts of N. nucifera leaves to inhibit angiogenesis in vivo was determined using the CAM assay. The numbers of blood vessel branch points significantly increased upon VEGF treatment (Fig. 3), compared with the PBS-treated control group (Fig. 3). However, treatment with both the water and methanol leaf extracts of N. nucifera significantly suppressed VEGF-induced angiogenesis in a dose-dependent manner (Fig. 3). In strong support of these results, it was previously found that alliin from garlic displayed inhibitory activity towards fibroblast growth factor-2-induced tube formation and in vivo CAM angiogenesis [29]. Alliin was also found to inhibit VEGF-induced angiogenesis in the CAM [29].

Bottom Line: In this study, N. nucifera leaf extracts displayed potent antioxidant and inhibitory effects on VEGF-induced angiogenesis.N. nucifera exerted an inhibitory effect on VEGF-induced proliferation and tube formation, as well as CAM angiogenesis in vivo.Moreover, N. nucifera leaf extracts significantly blocked VEGF-induced ROS production in HUVECs, confirming their possible anti-angiogenic mechanism.

View Article: PubMed Central - PubMed

Affiliation: Department of Food Science and Technology, Yeungnam University, Gyeongsan, Gyeongbuk, 712-749, Republic of Korea.

ABSTRACT
Nelumbo nucifera Gaertn (Nymphaeaceae) has long been used as a traditional herb in Chinese, Japanese, Indian, and Korean medicinal practices since prehistoric times and flourishes today as the primary form of medicine. This study reports for the first time the potent ability of N. nucifera leaf extracts to inhibit vascular endothelial growth factor (VEGF)-induced angiogenesis in vitro and in vivo, as well as their antioxidant efficacy in various scavenging models and an analysis of their chemical composition. In vivo anti-angiogenic activity was evaluated in a chick chorioallantoic membrane (CAM) model using fertilized chicken eggs, in human umbilical vein endothelial cells (HUVECs) by using cell viability, cell proliferation and tube formation assays, and by determining intracellular reactive oxygen species (ROS) in vitro. The antioxidant efficacy of N. nucifera leaf extracts was determined in various scavenging models, including total phenolic and flavonoid content. The chemical composition of N. nucifera leaf extracts was determined by GC-MS analysis, which revealed the presence of different phytochemicals. The IC50 values for the DPPH radical scavenging activities of water and methanol extracts were found to be 1699.47 and 514.36 μg ml(-1), and their total phenolic and flavonoid contents were 85.01 ± 2.32 and 147.63 ± 2.23 mg GAE g dry mass(-1) and 35.38 ± 1.32 and 41.86 ± 1.07 mg QA g dry mass(-1), respectively. N. nucifera leaf extracts (10-100 μg ml(-1)) exhibited significant dose-dependent inhibition of VEGF-induced angiogenesis, as well as VEGF-induced proliferation and tube formation in HUVECs. In this study, N. nucifera leaf extracts displayed potent antioxidant and inhibitory effects on VEGF-induced angiogenesis. N. nucifera exerted an inhibitory effect on VEGF-induced proliferation and tube formation, as well as CAM angiogenesis in vivo. Moreover, N. nucifera leaf extracts significantly blocked VEGF-induced ROS production in HUVECs, confirming their possible anti-angiogenic mechanism.

Show MeSH
Related in: MedlinePlus