title: Molecular mechanism of inhibition of the abnormal proliferation of human umbilical vein endothelial cells by hydroxysafflor-yellow A.
abstract: Context It has been found that hydroxysafflor-yellow A (HSYA) inhibits angiogenesis and the proliferation of abnormal human umbilical vein endothelial cells (HUVECs) in our previous study; however, the mechanism is still unclear. Objective This study investigates the mechanisms of HSYA inhibiting abnormal proliferation of HUVECs through detecting the expression of vascular endothelial growth factor (VEGF) and its receptor (KDR), and the protein expression in the Ras-Raf-MEK-ERK-signalling pathway. Materials and methods HepG2 cell cultural supernatant was used to culture HUVECs to make promote abnormal proliferation, and HSYA was added into the medium. The expression of VEGF, KDR, c-myc, N-ras and NF-κB1 in abnormal HUVEC was detected by RT-qPCR and ELISA at the mRNA and protein levels. Protein expression of ERK signal pathway was measured by Western blot. Results Compared with the abnormal proliferation of HUVECs without any treatment, HSYA inhibited the expression of VEGF and KDR in vitro. Similarly, the protein expression of Ras, p-raf, p-ERK and p-p38MARK in the abnormal HUVECs was reduced when they were treated by HSYA, especially in p-ERK, yet the total raf, ERK, p38MAPK and Akt were not changed whether HSYA existed or not. HSYA could also inhibit the expression of c-myc, N-ras, and NF-κB1. Conclusion When the abnormal HUVECs were treated with HSYA, the low expression of VEGF and KDR reduced the expression of oncogene and transcription factor through the Ras-Raf-MEK-ERK1/2 pathway of the MAPK family. This resulted in inhibiting the abnormal proliferation of HUVECs and angiogenesis.
authors.affiliation: a School of Basic Medicine, Beijing University of Chinese Medicine , Beijing , China ;
authors.forename: Ji
authors.initials: J
authors.lastname: Wang
authors.affiliation: a School of Basic Medicine, Beijing University of Chinese Medicine , Beijing , China ;
authors.forename: Jingjing
authors.initials: J
authors.lastname: Wang
authors.affiliation: a School of Basic Medicine, Beijing University of Chinese Medicine , Beijing , China ;
authors.forename: Xixi
authors.initials: X
authors.lastname: Wang
authors.affiliation: a School of Basic Medicine, Beijing University of Chinese Medicine , Beijing , China ;
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: a School of Basic Medicine, Beijing University of Chinese Medicine , Beijing , China ;
authors.forename: Jinghong
authors.initials: J
authors.lastname: Hu
authors.affiliation: a School of Basic Medicine, Beijing University of Chinese Medicine , Beijing , China ;
authors.forename: Xue
authors.initials: X
authors.lastname: Yu
authors.affiliation: a School of Basic Medicine, Beijing University of Chinese Medicine , Beijing , China ;
authors.forename: Yingying
authors.initials: Y
authors.lastname: Xu
authors.affiliation: b Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences , Beijing , China ;
authors.forename: Xuyan
authors.initials: X
authors.lastname: Niu
authors.affiliation: c Zhejiang Provincial Key Laboratory of Applied Enzymology , Yangtze Delta Region Institute of Tsinghua University , Jiaxing , China.
authors.forename: Zong
authors.initials: Z
authors.lastname: Lin
authors.affiliation: a School of Basic Medicine, Beijing University of Chinese Medicine , Beijing , China ;
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: a School of Basic Medicine, Beijing University of Chinese Medicine , Beijing , China ;
authors.forename: Xin
authors.initials: X
authors.lastname: Zhang
authors.affiliation: a School of Basic Medicine, Beijing University of Chinese Medicine , Beijing , China ;
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects*; D002599:Chalcone / Q000031:analogs & derivatives* / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D056945:Hep G2 Cells; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D052002:NF-kappa B p50 Subunit / Q000235:genetics / Q000378:metabolism; D009389:Neovascularization, Pathologic; D019899:Paracrine Communication; D010766:Phosphorylation; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D011809:Quinones / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism; D048490:raf Kinases / Q000235:genetics / Q000378:metabolism; D018631:ras Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D020533:Angiogenesis Inhibitors; D017077:Culture Media, Conditioned; C489427:MYC protein, human; D052002:NF-kappa B p50 Subunit; C489548:NFKB1 protein, human; D016271:Proto-Oncogene Proteins c-myc; D011809:Quinones; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C085278:hydroxysafflor yellow A; D002599:Chalcone; C000588866:KDR protein, human; D040301:Vascular Endothelial Growth Factor Receptor-2; D048490:raf Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D020929:Mitogen-Activated Protein Kinase Kinases; D018631:ras Proteins
keywords: Angiogenesis; MAPK signalling pathway; VEGF