NOD2 stimulation enhances the innate immunity against Mycobacterium tuberculosis in human alveolar macrophages / 国际检验医学杂志

Objective To evaluate the role of nucleotide‐binding oligomerization domain 2(NOD2) stimulation in innate immuni‐ty against M ycobacterium tuberculosis .Methods Plate counting as used to evaluate the effect of resisting M ycobacterium tubercu‐losis in human alveolar macrophages .Intracellular NOD2 expression were detected by flow cytometry .Quantitative real‐time PCR was performed to determine the NOD2 ,inducible nitric oxide synthase(iNOS) ,and DEF4B mRNA expression levels using the com‐parative threshold cycle method of relative quantitation .Reactive oxygen species(ROS) were detected by the DFCH probe .Results NOD2 stimulation enhanced the control of intracellular mycobacterial growth in human alveolar macrophages .Although ROS con‐centration did not changed ,the secretion of Nitro Oxygen and the expression of cathelicidin DEFB4 were significantly increased fol‐lowing NOD2 stimulation in human alveolar macrophages .Conclusion NOD2 stimulation may be involved in the early innate con‐trol of Mycobacterium tuberculosis primary infections inducing the generation of Nitro Oxygen and the peptides cathelicidin DEFB4 .

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

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