Role of nucleotide binding oligomerization domain receptor 1 as a new innate immune recognition receptor in activation of vascular smooth muscle cells / 中国组织工程研究

ABSTRACT

AIM: Nucleotide binding oligomerization domain receptor 1 (NOD1) is a recently identified intracellular pathogen pattern recognition receptor of innate immunity. This study was designed to explore the role of NOD1-mediated innate immune signal pathway in the activation of vascular smooth muscle cells (VSMCs), and investigate the effect of peptidoglycan (PGN) on the activation and expression of NOD1 in human VSMCs. METHODS: The experiment was performed in the central laboratory of the Second Affiliated Hospital of Dalian Medical University from June 2006 to March 2007.①Subject: Human coronary artery VSMCs were purchased from Cambrex company.② Methods: Human coronary artery VSMCs were cultured in vitro, and stimulated with NOD1 agonist PGN (10 mg/L) for 0, 3, 6 and 24 hours. ③ Evaluation: The mRNA expression of NOD1 in VSMCs was measured by real time quantitative reverse transcription-polymerase chain reaction. The concentration in the culture supernatants of interleukin-8 (IL-8) and tumor necrosis factor-? (TNF-?) was determined by enzyme linked immunosorbent assay. RESULTS: ①Human VSMCs constitutively expressed a low level of NOD1 at resting condition. Upon PGN stimulation, the expression of NOD1 mRNA was up-regulated in VSMCs, from 0.164?0.005 to 0.231?0.027 (P