Research On Inhibition Effect Of Methylprednisolone On Activation Of NF-κB And Expression Of Interleukin 6,Interleukin 17 In Peripheral Blood Mononuclear Cell In Acute Stage Of Kawasaki Disease

Obsjective: |Kawasaki disease(KD) is an acute vasculitis disease of children, manife- sted by fever and signs of mucocutaneous inflammation, Its etiology is unkown, but it involves an obvious immunological regulation dysfuction in the acute stage. Glucoco- rtisteroids(GC) is no longer a ban to treat KD.This study aimed to investigate the inhibinition effect of methylprednisolone on nuclear-factor-kappaB (NF-κB) activa- tion in peripheral blood mononuclear cells(PBMCs) and the expression of interleukin-6 (IL-6),IL-17 in the culture supernatant in the acute stage of KD so that to explore the antiinflammatory mechanism of GC in the treatment of KD and to explore the pathogenesis of KD.Methods:PBMCs were isolated and purified from the blood of 20 KD children and 20 healthy children by Ficoll density gradient centrifugation and then divided the sa- mples into four groups and then cultured for 48 hours. The first group was cultured n- aturally;The second group was only stimulated by PMA (Phorbol 12-Myristate 13-a- cetete); The third group was cultured with PMA﹢methylprednisolone (MP, final con- centration 250μg/mL);The last group was cultured with PMA﹢dexamethasone(DEX, final concentration 5μg/mL).The percentage of NF-κB activated in PBMCs was dete- cted by immunocytochemical straining, and the expression level of IL-6 or IL-17 was measured by enzyme-linked immunosorbentassay (ELISA).Results:1. Under natural culturing and PMA stimulating, the expression level of IL-17,IL-6 and the activation level of NF-κB in KD patients were markedly increased when compared to healthy subjects (P<0.01).2. In acute KD patients, the activation level of NF-κB and the expression of IL-17,IL-6 in MP group and in DEX group were markedly decreased when compared to that in PMA group (P<0.01).3. In KD patients, there was no evident difference of the activation level of NF-κB and the expression level of IL-17,IL-6 between MP group and DEX group (P> 0.05). 4. After PMA stimulation, there was obvious positive linear correlation between the activation level of NF-κB and the expression level of IL-17,IL-6 in KD patients (r =0.587,p<0.01, r=0.453,p<0.01).5. After PMA stimulation, there was obvious positive linear correlation between the expression level of IL-17 and IL-6 in KD patients (r=0.738,p<0.01).Conclusion:1. The high secretion of IL-17,IL-6 by PBMCs and activation of NF-κB in PBMCs play important roles in endothelial immunity damage of acute KD.2. The antiinflammatory mechanism of MP plays an evident role in inhibiting activation of NF-κB and high secretion of IL-17,IL-6 in PBMCs in acute KD.3. There was no evident difference of anti-inflammatory effect between MP and DEX in acute KD patients.