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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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Effects of water and methanol extracts of N. nucifera leaves on cell viability and VEGF-induced proliferation.(a) Cell viability: HUVECs were treated with various concentrations of extracts. After 48 h, cell viability was measured using the MTT assay. *: p<0.05 compared with untreated control (0 μg ml−1). (b) VEGF-induced proliferation: HUVECs were co-treated with various concentrations of extracts and VEGF for 48 h. The number of viable cells was measured by using the MTT assay. *: p<0.05 compared with untreated control; #: p<0.05 compared the VEGF-treated group. All tested concentrations (10, 50, and 100 μg ml−1) of the water as well as the methanol extracts displayed statistically significant differences with respect to each other.
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pone.0118552.g001: Effects of water and methanol extracts of N. nucifera leaves on cell viability and VEGF-induced proliferation.(a) Cell viability: HUVECs were treated with various concentrations of extracts. After 48 h, cell viability was measured using the MTT assay. *: p<0.05 compared with untreated control (0 μg ml−1). (b) VEGF-induced proliferation: HUVECs were co-treated with various concentrations of extracts and VEGF for 48 h. The number of viable cells was measured by using the MTT assay. *: p<0.05 compared with untreated control; #: p<0.05 compared the VEGF-treated group. All tested concentrations (10, 50, and 100 μg ml−1) of the water as well as the methanol extracts displayed statistically significant differences with respect to each other.

Mentions: The cytotoxic effects of N. nucifera leaf extracts were examined using the MTT assay to determine the effective concentrations required for the treatment. Exposing HUVECs to 1 to 1,000 μg ml−1 of the water extract for 48 h did not reduce cell viability; however, the viability of cells exposed to 500 and 1,000 μg ml−1 of the methanol extract for 48 h was reduced to 25% compared with the viability of control cells (Fig. 1A). These findings indicate that N. nucifera leaf extracts did not affect the viability of HUVEC cells at concentration lower than 100 μg ml−1 (Fig. 1A).


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)

Effects of water and methanol extracts of N. nucifera leaves on cell viability and VEGF-induced proliferation.(a) Cell viability: HUVECs were treated with various concentrations of extracts. After 48 h, cell viability was measured using the MTT assay. *: p<0.05 compared with untreated control (0 μg ml−1). (b) VEGF-induced proliferation: HUVECs were co-treated with various concentrations of extracts and VEGF for 48 h. The number of viable cells was measured by using the MTT assay. *: p<0.05 compared with untreated control; #: p<0.05 compared the VEGF-treated group. All tested concentrations (10, 50, and 100 μg ml−1) of the water as well as the methanol extracts displayed 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.g001: Effects of water and methanol extracts of N. nucifera leaves on cell viability and VEGF-induced proliferation.(a) Cell viability: HUVECs were treated with various concentrations of extracts. After 48 h, cell viability was measured using the MTT assay. *: p<0.05 compared with untreated control (0 μg ml−1). (b) VEGF-induced proliferation: HUVECs were co-treated with various concentrations of extracts and VEGF for 48 h. The number of viable cells was measured by using the MTT assay. *: p<0.05 compared with untreated control; #: p<0.05 compared the VEGF-treated group. All tested concentrations (10, 50, and 100 μg ml−1) of the water as well as the methanol extracts displayed statistically significant differences with respect to each other.
Mentions: The cytotoxic effects of N. nucifera leaf extracts were examined using the MTT assay to determine the effective concentrations required for the treatment. Exposing HUVECs to 1 to 1,000 μg ml−1 of the water extract for 48 h did not reduce cell viability; however, the viability of cells exposed to 500 and 1,000 μg ml−1 of the methanol extract for 48 h was reduced to 25% compared with the viability of control cells (Fig. 1A). These findings indicate that N. nucifera leaf extracts did not affect the viability of HUVEC cells at concentration lower than 100 μg ml−1 (Fig. 1A).

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