关键词: NOD1, 肝窦内皮细胞, 二氨基庚二酸, 信号通路 

Abstract: Objective To investigate the effects of the immune response mediated by NOD1 receptor in liver sinusoidal cells ( LSECs ) of mice. Method Firstly, We used the method of the collagenase perfusion, percoll density gradient centrifugation and anti-LSEC MicroBeads separation and purification to obtain the high purity LSECs. The constitutive expression of NOD1 in LSEC was detected by real-time RT-PCR. Then the LSECs were stimulated with the diaminopimelic acid ( C12-ieDAP ) , the cells and cell supernatant were collected for 6 hours and 20 hours. Total RNA was isolated from LSECs for detecting the mRNA of NOD1, RIP2, IL-6, TNF-α, CXCL-1, CXCL-2, CXCL-9, CXCL-10, CCL-2 and CCL-5 by qRT-PCR. The cytokines ( IFN-α, IFN-β, IFN-γ, TNF-α and IL-6 ) in the cells supernatant were detected by quantitative ELISA. Result Our data showed that NOD1 mRNA was highly expressed at baseline in primary LSEC, and that the activation of LSEC by the NOD1 ligand C12-ieDAP could induce up-regulation of NOD1 mRNA and its connector RIP2 mRNA. The mRNA levels of cytokines IL-6 and chemokines CXCL-1, CXCL-2, CXCL-9, CCL-2 and CCL-5 were up-regulated. Stimulation of NOD1 ligand C12-ieDAP with the concentration of 10 μg / mL induced the production of IFN-γ and IL-6 effector molecules, but not other cytokines. Conclusion In conclusion, NOD1 was functionally expressed in LSEC, could be activated by its ligand, and mediated the activation of NF-κB signaling pathway to produce IL-6 and IFN-γ effector molecules as well as a series of cellular inflammatory cytokines and chemokines.

Key words: NOD1, liver sinusoidal endothelial cell, diaminopimelic acid, signaling pathway