Chlorotyrosine promotes human aortic smooth muscle cell migration through increasing superoxide anion production and ERK1/2 activation

Abstract

Chlorotyrosine is an oxidative product of hypochlorous acid and L-tyrosine, and is considered as a biomarker for oxidative stress and cardiovascular disease. However, it is not clear whether chlorotyrosine could directly contribute to vascular pathogenesis. In this study, we investigated the effect and potential mechanisms of chlorotyrosine on human aortic smooth muscle cell (AoSMC) migration. With Boyden chamber and wound healing assays, chlorotyrosine significantly increased AoSMC migration in a concentration-and time-dependent manner. In addition, chlorotyrosine significantly increased the expression of several key molecules related to cell migration including PDGF receptor-B (PDGFR-B), matrix metalloproteinases (MMP-1 and MMP-2) and integrins (α3, αV, and β3) in AoSMC at both mRNA and protein levels. Furthermore, chlorotyrosine also increased superoxide anion generation in AoSMC with the fluorescent dye dihydroethidium (DHE) staining. Activation of mitogen-activated protein kinases (MAPKs) was analyzed with Bio-Plex Luminex immunoassay and western blotting. Chlorotyrosine induced a transient phosphorylation of ERK1/2, but not JNK and p38 MAPKs. Antioxidants including selenomethionine (SeMet) and Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) as well as ERK1/2 inhibitor PD98059 effectively blocked chlorotyrosine-induced AoSMC migration. Thus, these findings demonstrate new biological functions of chlorotyrosine in human SMC migration, which may play a crucial role in the vascular lesion formation.

Interstitial flow promotes vascular fibroblast, myofibroblast, and smooth muscle cell motility in 3-D collagen I via upregulation of MMP-1

“…Twodimensional shear stress suppresses SMC migration in Matrigel via the downregulation of MMP-2 activity (8,28). The upregulation of MMP-1 and -2 are responsible for chlorotyrosine-induced human aortic SMC migration (24). Other recent studies reported that fluid shear stress modulated endothelial cell invasion into 3-D collagen matrixes through MMP-2 activation and that collagen I but not Matrigel matrixes form an MMP-dependent barrier to ovarian cancer cell penetration (13,44).…”

Lactosylceramide promotes cell migration and proliferation through activation of ERK1/2 in human aortic smooth muscle cells

“…The MAPK phosphorylation state of AoSMC lysates was analyzed by Bio-Plex phosphoprotein and total target assays (Bio-Rad, Hercules, CA) as described in our previous publications (23,24). Briefly, serum-starved AoSMCs were treated with LacCer (10 M), and cell lysates were collected at different time points of 0, 5, 7, 10, 15, 20, 30, 45, and 60 min.…”

“…Serum-starved AoSMCs were treated with LacCer (10 M) or DMSO (0.1%) for 24 h, and total RNA was isolated for real-time PCR analysis, as described in our previous study (24). Human PDGFR-B, MMPs, and integrins as well as ␤-actin primers were designed using Beacon Designer software.…”

PEG-nanolized ultrasmall selenium nanoparticles overcome drug resistance in hepatocellular carcinoma HepG2 cells through induction of mitochondria dysfunction

“…The PEG200 spectrum showed characteristic bands of specific functional groups, Abbreviations: Se, selenium; TEM, transmission electron microscopy; SEM-EDX, scanning electron microscope energy-dispersive X-ray spectroscopy; PEG-SeNPs, polyethylene-glycol-nanolized selenium nanoparticles. such as the bands appearing at 2874.2 cm -1 assigned to the -CH group 23 and the band at 1103.9 cm -1 assigned to the -C-O-C group. 24 These two characteristic bands and some other bands near 1103.9 cm -1 also appeared in the PEG-SeNP spectrum, indicating the successful conjugation of PEG to the surface of the SeNPs.…”