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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 tube formation in HUVECs.(a) HUVECs (5 x 104 cells) were plated on wells that had been previously coated with 40 μL of growth factor-reduced Matrigel basement membrane matrix. Cells were then treated with extracts in the presence of VEGF (20 ng ml−1). After 14 h, cells were photographed with a digital camera under a phase contrast microscope at 40x magnification. (b)The bar graph represents the relative area covered by the tube network, and data are shown as the mean ± SD. *: p<0.05 compared with untreated control; #: p<0.05 compared with VEGF-treated group.
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pone.0118552.g002: Inhibitory effects of water and methanol extracts of N. nucifera leaves on tube formation in HUVECs.(a) HUVECs (5 x 104 cells) were plated on wells that had been previously coated with 40 μL of growth factor-reduced Matrigel basement membrane matrix. Cells were then treated with extracts in the presence of VEGF (20 ng ml−1). After 14 h, cells were photographed with a digital camera under a phase contrast microscope at 40x magnification. (b)The bar graph represents the relative area covered by the tube network, and data are shown as the mean ± SD. *: p<0.05 compared with untreated control; #: p<0.05 compared with VEGF-treated group.

Mentions: Furthermore, we examined the effects of both extracts on HUVEC tube formation. VEGF treatment significantly enhanced tube formation, which was inhibited by the water and methanol leaf extracts of N. nucifera in a dose-dependent manner (Fig. 2). According to the relevant literature, the inhibitory effects of many dietary polyphenols, including green tea polyphenols and mushroom polyphenols, on angiogenesis, metastasis, and tumor growth are believed to be mediated through the regulation of VEGFR-dependent signaling pathways [28]. Our results suggest that N. nucifera leaf extracts significantly blocked VEGF-induced angiogenesis in vitro. Moreover, the anti-angiogenic activities of the water and methanol leaf extracts were in accordance with their antioxidant activities, which may be attributed to the presence of large amounts of total phenolics and flavonoids in both the extracts.


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 tube formation in HUVECs.(a) HUVECs (5 x 104 cells) were plated on wells that had been previously coated with 40 μL of growth factor-reduced Matrigel basement membrane matrix. Cells were then treated with extracts in the presence of VEGF (20 ng ml−1). After 14 h, cells were photographed with a digital camera under a phase contrast microscope at 40x magnification. (b)The bar graph represents the relative area covered by the tube network, and data are shown as the mean ± SD. *: p<0.05 compared with untreated control; #: p<0.05 compared with VEGF-treated group.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0118552.g002: Inhibitory effects of water and methanol extracts of N. nucifera leaves on tube formation in HUVECs.(a) HUVECs (5 x 104 cells) were plated on wells that had been previously coated with 40 μL of growth factor-reduced Matrigel basement membrane matrix. Cells were then treated with extracts in the presence of VEGF (20 ng ml−1). After 14 h, cells were photographed with a digital camera under a phase contrast microscope at 40x magnification. (b)The bar graph represents the relative area covered by the tube network, and data are shown as the mean ± SD. *: p<0.05 compared with untreated control; #: p<0.05 compared with VEGF-treated group.
Mentions: Furthermore, we examined the effects of both extracts on HUVEC tube formation. VEGF treatment significantly enhanced tube formation, which was inhibited by the water and methanol leaf extracts of N. nucifera in a dose-dependent manner (Fig. 2). According to the relevant literature, the inhibitory effects of many dietary polyphenols, including green tea polyphenols and mushroom polyphenols, on angiogenesis, metastasis, and tumor growth are believed to be mediated through the regulation of VEGFR-dependent signaling pathways [28]. Our results suggest that N. nucifera leaf extracts significantly blocked VEGF-induced angiogenesis in vitro. Moreover, the anti-angiogenic activities of the water and methanol leaf extracts were in accordance with their antioxidant activities, which may be attributed to the presence of large amounts of total phenolics and flavonoids in both the extracts.

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