N-benzyl-5-phenyl-1H-pyrazole-3-carboxamide Promotes Vascular Endothelial Cell Angiogenesis and Migration in the Absence of Serum and FGF-2

Overview

Journal Acta Pharmacol Sin

Specialty Pharmacology

Date 2011 Feb 5

PMID 21293473

Citations 4

Authors

Hai-Yan Zhang

Le Su

Bin Huang

Jing Zhao

Bao-Xiang Zhao

Shang-li Zhang

Jun-Ying Miao

Affiliations

Soon will be listed here.

Abstract

Aim: To investigate the effect of N-benzyl-5-phenyl-1H-pyrazole-3-carboxamide (BPC) on angiogenesis in human umbilical vein endothelial cells (HUVECs).

Methods: Capillary-like tube formation on matrigel and cell migration analyses were performed in the absence of serum and fibroblast growth factor (FGF-2). Reactive oxygen species (ROS) were measured using a fluorescent probe, 2', 7'- dichlorodihydrofluorescein (DCHF). The nitric oxide (NO) production of HUVECs was examined using a NO detection kit. Morphological observation under a phase contrast microscope, a viability assay using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl-tetrazolium (MTT) and a lactate dehydrogenase (LDH) activity analysis by a detection kit were performed to evaluate the toxicity of BPC on HUVECs in the presence of serum and FGF-2. The level of hypoxia-inducible factor 1α (HIF-1α) and the release of vascular endothelial growth factor (VEGF) were measured by Western blot and ELISA, respectively.

Results: In the absence of serum and FGF-2, cells treated with BPC (5-20 μmol/L) rapidly aligned with one another and formed tube-like structures within 12 h. In the presence of serum and FGF-2, cells treated with BPC for 24, 48 and 72 h had no changes in morphology, viability or LDH release compared with the control group. Cell migration in the BPC-treated group was significantly increased compared with the control group. During this process, NO production and ROS level were elevated dramatically, and the levels of HIF-1α and VEGF were increased dependent on the generation of ROS.

Conclusion: BPC most effectively promoted angiogenesis and migration in HUVECs in the absence of FGF-2 and serum.

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