Open AccessTransforming growth factor–β1 mediated CHST11 and CHSY1 mRNA expression is ROS dependent in vascular smooth muscle cells
Author(s) -
Mohamed Raafat,
Dayati Parisa,
Mehr Reyhaneh Niayesh,
Kamato Danielle,
Seif Faezeh,
BabaahmadiRezaei Hossein,
Little Peter J.
Publication year - 2018
Publication title -
journal of cell communication and signaling
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.329
H-Index - 44
eISSN - 1873-961X
pISSN - 1873-9601
DOI - 10.1007/s12079-018-0495-x
Subject(s) - phosphorylation , microbiology and biotechnology , vascular smooth muscle , signal transduction , kinase , transforming growth factor , chemistry , chondroitin sulfate , biochemistry , biology , glycosaminoglycan , endocrinology , smooth muscle
Transforming growth factor (TGF)‐β1 mediates glycosaminoglycan (GAG) chain hyperelongation on secreted proteoglycans and these modifications are associated with increased lipid binding in the vessel wall and the development of atherosclerosis. In vascular smooth muscle cells (VSMCs), TGF‐β1 regulated GAG elongation via extracellular signal‐regulated kinase (ERK) and p38 as well as Smad2 linker region phosphorylation. In this study, our aim was to identify the TGF‐β1 mediated signalling pathway involving reactive oxygen species (ROS) and Smad2 linker region phosphorylation that regulate the mRNA expression of GAG synthesizing enzymes, chondroitin 4‐O‐sulfotransferase 1 (CHST11) and chondroitin sulfate synthase 1 (CHSY1) which are the rate limiting enzymes involved in GAG chain elongation. Signalling molecules were assessed by western blotting, quantitative real‐time PCR was used for analysis of gene expression and intracellular ROS level was measured by a fluorescence based assay. TGF‐β1 induced ROS production in VSMCs. Nicotinamide adenine dinucleotide phosphate oxidase (Nox) inhibitors, diphenyleneiodonium (DPI) and apocynin blocked TGF‐β1 mediated Smad2 linker region phosphorylation. TGF‐β1 treatment increased the mRNA levels of CHST11 and CHSY1. Pharmacological inhibition of Nox blocked TGF‐β1 mediated mitogen activated protein kinases (MAPKs) phosphorylation and TGF‐β1 stimulated CHST11 and CHSY1 mRNA expression. These findings demonstrated that TGF‐β1 mediated expression of CHST11 and CHSY1 can occur via Nox‐dependent pathways and Smad2 linker region phosphorylation.