Synergistic Effect of Nucleotide binding Oligomerization Domain 2 and Toll-like Receptor Agonists on Proliferation and Release of Proinflammatory Cytokines in Vascular Smooth Muscle Cells / 中华高血压杂志

ABSTRACT

Objective To investigate the expression of nucleotide-binding oligomerization domain 2(NOD2),an intracelluar pathogen pattern recognition receptor,and Toll like receptor(TLR) 2,4 in human vascular smooth muscle cells(VSMC),and its effect on production of proinflammatory cytokines in VSMC.Methods Human coronary artery smooth muscle cells were in vitro stimulated with NOD2 agonist Muramyl dipeptide(MDP),TLR2 agonist Pam3CSK4(PAM3)and TLR4 agonist lipopolysaccharides(LPS) alone or MDP in cocultured with either PAM3 or LPS.The mRNA expression of NOD2 and fibroblast growth factor-2(FGF-2) were measured by real time RT-PCR.The concentration in the culture supernatants of interleukin-8(IL-8) and tumor necrosis factor-?(TNF-?) was determined by ELISA.VSMC proliferation was analyzed by the MTT assay.Results MDP up-regulate the expression of NOD2 mRNA in VSMC in a time-dependent manner(0 h:0.028?0.001;3 h:0.045?0.002;6 h:0.053?0.002;24 h:0.162?0.013).It up-regulate the expression of FGF-2 mRNA(MDP 9.3?0.4 vs control 7.4?0.2;P