Effects Of Aspirin On The Activation Of Nuclear Factor-kappa B And The Expression Of Interleukin-8 Induced By Tumor Necrosis Factor-αin Peripheral Blood Mononuclear Cells In Patients With Kawasaki Disease

Objective:Aspirin is the most common antiinflammatory drug used in the acute stage of Kawasaki disease(KD), but it's antiinflammatory mechanism is not yet very clear .In this study, we used tumor necrosis factor-α(TNF-α) to activate nuclear factor-kappa B(NF-κB) in the peripheral blood mononuclear cells(PBMCs) of KD and then evaluated the inhibition effect of aspirin on the activation of NF-κB and the subsequent expression of interleukin-8(IL-8). By this way, we hope that we might prove that the main antiinflammatory mechanism of aspirin in the treatment of acute Kawasaki disease(KD) is associated with NF-κB activation.Methods: PBMCs were isolated and purified from blood of twenty KD children by density gradient centrifugation and then divided into five groups and cultured for 2 hours. The first group was cultured naturally. The second group was only stimulated by TNF-αOther groups were"TNF-α+aspirin groups"which were treated with different concentration of aspirin and stimulated by TNF-α. Activation of NF-κB in PBMCs were detected by immunocytochemical straining, and enzyme-linked immuosorbent assy(ELISA)was used to measure the concentration of IL-8 in the supernanant.Results:.(1) Comparing with the group under natural culturing(50.59±9.02%,619.04±256.18pg/ml), the activition level of NF-κB (79.86±8.24%)and the expression level of IL-8(1324.53±300.06pg/ml)in TNF-αgroup were all much higher (P=0.000).(2) The NF-κB activation levels(70.75±7.36%,51.80±7.97%)in 5,10mmol/L aspirin+ TNF-αgroup were all inhibited significantly(P=0.002, P=0.000), and the expression of IL-8(1016.03±268.75 pg/ml,715.15±219.80pg/ml) in 5 or 10mmol/L aspirin+ TNF-αgroup were inhibited significantly too(P=0.003, P=0.000), while inhibition of NF-κB activation (75.30±8.62%)and IL-8 expression(1176.69±309.57pg/ml) in 2mmol/L aspirin+ TNF-αgroup were not evident(P=0.193,P=0.245).(3) There was obvious positive correlation between the activation of NF-κB in PBMCs and the expression of IL-8 in the cultured supernatant(r=0.764, P=0.000).Conclusion: Aspirin can inhibit the activation of NF-κB in PBMCs induced by TNF-αin the acute stage of KD, and aspirin can also inhibit the expression of IL-8 through the same pathway. Results of our study suggest that the inhibitory effect on the activation of NF-κB in PBMCs is the most important antiinflammatory mechanism of aspirin in the treatment of acute stage of KD.