Experimental study on the NF-κB-mediated matrix metalloproteinase-9 expression in peripheral blood mononuclear cells in patients with Kawasaki disease

TAN Cheng; HAN Qin; ZHU Chun-liang; LI Zhe-qian; LIU Li-sha

doi:10.3969/j.issn.1002-266X.2015.06.004

Abstract：Objective To explore the activation of nuclear factor-kappaB ( NF-κB ) and matrix metalloproteases 9 (MMP-9) in peripheral blood mononuclear cells (PBMCs)in children with Kawasaki disease and the relationship between NF-κB and MMP-9.Methods PBMCs were isolated and purified from blood of fifty-six children including 21children with KD, 18 children with general inflammatory and 17 normal ones by density gradient centrifugation ,and were cultured for one hour after cell concentration adjusted to 106/mL.Then both KD group and Control group were divided into three groups ,the first group was cultured naturally , the second one was stimulated by phorbol 12-myristate 13-acetate(PMA), and the third one was stimulated by PMA and pyrrolidine dithiocarbamate ( PDTC) .PBMCs were cultured for 2 h.The mRNA and pro-tein levels of NF-κB p65 in PBMCs in each groups were measured by RT-PCR and Western blot , and the protein levels of MMP-9 were measured by ELISA .Results The mRNA and protein levels of NF-κB p65 and MMP-9 in KD group were much higher than that in other groups;Stimulating PBMCs by PMA ,the mRNA and protein levels of NF-κB p65 and MMP-9 were markedly increased in each group , and there was a significant difference in each group (P<0.05); Using PDTC can reduce the activation of NF-κB in PBMCs, and the protein levels of MMP-9 were significantly reduced by using PDTC in KD group.Conclusions The activity of NF-κB and MMP-9 in PBMCs in patients with acute KD is distinctly increased;The specific inhibitor of NF-κB can inhibit it's activation and the expression of MMP-9 obviously.It suggested that the NF-κB pathway can lead to the activation of MMP-9 in acute KD, and proper application of specific inhibitor of NF-κB may be a new choice for treating KD .