Effect Of Tanshinone ⅡA The Activation Of Nuclear Factor-kappa B And The Expression Of Inflammatory Cytokines In Peripheral Blood Mononuclear Cells (PBMCS) Of The Acute Phase Of Kawasaki Disease

Objective: To explore the inhibition effect of Tanshinone IIA (TanⅡA) on the activation of nuclear factor-kappa B (NF-κB) and the expression ofTNF-α、IL-1β、IL-8in peripheral blood mononuclear cells (PBMCs) of theacute phase of Kawasaki disease(KD) and to explore the pathogenesis ofKawasaki disease(KD) and to explore the anti-inflammatory effect ofTanshinone IIA and compare with Acetylsalicylic acid(ASA).Methods: PBMCs were isolated and purified from peripheral blood of20KD children and20healthy children by density gradient centrifugation.In every sample PBMCs were divided into six groups. The first group wascultured naturally, the second one was stimulated by phorbol12-myristate-13-acetate (PMA), the third one was stimulated by PMA andASA, other groups were stimulated by PMA and TanshinoneIIA which wastreated with different concentrations of TanⅡA (the final concentration ofTan II A are1mg/l、3mg/l、10mg/l). Every group was cultured in Carbon dioxide incubator. Activation of NF-κB in PBMCs were determined byimmunocytochemical, and ELISA was used to measure the concentration ofTNF-α、IL-1β、IL-8in supernatant.Results: In the conditions of natural culture、 PMA to stimulateculture、 PMA+different concentrations of TanⅡA culture (the finalconcentration of Tan II A are1mg/l、3mg/l、10mg/l) and PMA+ASAculture，the activation of nuclear factor-kappa B(NF-κB)and the expressionof TNF-α、IL-1β、IL-8in KD group, was significant higher than controlgroup(p=0.000). Under PMA culturing, the activation of NF-κB wassignificantly higher than the black group both in KD and control group（p＜0.01，p＜0.05）; in supernatant of the two group, the expression ofTNF-α、IL-1β and IL-8was significantly higher than the blak group（p＜0.01）. The activation of NF-κB and the expression of TNF-α、IL-1β andIL-8in KD and control group,there was no difference between the PMA+final concentration of1mg/l Tan II A group and the PMA group (p＞0.05)；Under PMA+final concentration of3mg/l Tan II A culturing, the activationof NF-κB in KD and control group were significantly lower than the PMAgroup（p＜0.01，p＜0.05）, the expression of IL-1β and IL-8in KD andcontrol group,and the expression of TNF-α in KD group were significantlylower than the PMA group（p＜0.01）,while in supernatant of the controlgroup, the expression of TNF-α, there was no significantly decline（p＞0.05）. Under PMA+final concentration of10mg/l Tan II A culturing, the activation of NF-κB in KD and control group were significantly lower thanthe PMA group（p＜0.01，p＜0.05）, in supernatant of the KD and controlgroup the expression of TNF-α、IL-1β and IL-8were significantly lower too.and the PMA+final concentration of10mg/l Tan II A group was lower thanthe PMA+final concentration of3mg/l Tan II A group both in the KD andcontrol group. Under PMA+ASA culturing, the activation of NF-κB and theexpression of TNF-α、IL-1β and IL-8in KD and control group weresignificantly lower than PMA group,and significantly lower than the PMA+final concentration of10mg/l Tan II A group too, There was obviouspositive correlation between the activation of NF-κB in PBMCs and theexpression of TNF-α、IL-1β and IL-8in the cultured supernatant（r=0.817，p＜0.01；r=0.665，p＜0.05；r=0.782，p＜0.01）.Conclusion:①The activation of NF-κB enhanced and the expressionof TNF-α、IL-1β and IL-8increased in peripheral blood mononuclear cells(PBMCs) of the acute phase of Kawasaki disease(KD) involved in immuneinflammatory response and may be mediated immune vasacular injury.②The anti-inflammatory effect of Tan II A in peripheral blood mononuclearcells (PBMCs) of the acute phase of Kawasaki disease(KD) has adose-dependence, and the anti-inflammatory mechanism same as ASA:inhibit the activation of NF-κB and the subsequent expression of TNF-α、IL-1β and IL-8.