丹参酮ⅡA和阿司匹林对急性期川崎病患儿外周血单个核细胞NF-κB活化及炎性细胞因子表达影响的研究  

The Effects of Tanshinone ⅡA and Aspirin on Nuclear Factor-kappa B and Inflammatory Cytokines in Peripheral Blood Mononuclear cells of Kawasaki Disease

作　　者：汪燕[1] 李晓静[1] 于洁[2] 周敏[1] 黄成[1] 王雪梅[1] 

机构地区：[1]重庆医科大学附属成都市妇女儿童中心医院血液科,成都610091 [2]重庆医科大学附属儿童医院血液科,重庆400014

出　　处：《血栓与止血学》2012年第6期251-256,共6页Chinese Journal of Thrombosis and Hemostasis

基　　金：四川省卫生厅科研课题;编号:110061

摘　　要：目的观察丹参酮ⅡA(TanⅡA)和阿司匹林(ASP)对急性期川崎病(KD)患儿外周血单个核细胞(PB-MCs)核转录因子-κB(NF-κB)活化及肿瘤坏死因子-α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素-8(IL-8)表达的影响。探讨丹参酮ⅡA对KD患儿的抗炎作用机制,并与传统药物阿司匹林的作用进行比较。方法病例选择为确诊KD的治疗前住院患儿20例,同期门诊同龄健康体检儿童20例作健康对照组。分别取外周静脉血5 ml,采用Ficoll密度梯度离心法分离出静脉血中的单个核细胞,并将试验分为4组进行培养,即自然培养组(空白组)、12-肉豆蔻酰-13乙酸佛波酯(PMA)组、PMA+ASP组和PMA+TanⅡA组,免疫细胞化学法观察培养后各组细胞中NF-κB表达率,ELISA法检测各组培养上清液中TNF-α、IL-1β、IL-8的表达量。结果 KD各组中NF-κB的表达率及炎性细胞因子TNF-α、IL-1β、IL-8的表达量均显著高于健康对照组(P=0.000、P=0.000、P=0.000、P=0.000)。KD组中PMA组NF-κB的表达率及炎性细胞因子TNF-α、IL-1β、IL-8的表达量显著高于空白组(P<0.01、P<0.01、P<0.01、P<0.01);PMA+ASP组和PMA+TanⅡA组NF-κB的表达率及炎性细胞因子TNF-α、IL-1β、IL-8的表达量均较PMA组显著降低(P<0.01、P<0.01、P<0.01、P<0.01);PMA+ASA组较PMA+TanⅡA组降低更明显(P<0.01、P<0.05、P<0.01、P<0.01)。NF-κB的表达率与TNF-α、IL-1β、IL-8的表达量呈正直线相关(r=0.842,P<0.01;r=0.898,P<0.01;r=0.801,P<0.01)。结论丹参酮ⅡA在KD急性期PBMCs中具有明显的抗炎作用,其机制可能与ASA相同,即通过抑制NF-κB的活化,进而抑制其下游TNF-α、IL-1β、IL-8的表达而实现。Objective To explore the effect of Tanshinone ⅡA （Tan Ⅱ A） and aspirin （ASP） on the mechanism of Kawasaki disease （ KD）, which effect on uclear factorkappa B （ NF- κB ） and TNF-α, IL- 1β, IL-8 in peripheral blood mononuclear cells （PBMCs） of KD. Methods 20 cases of hospitalized children with KD were involved, controled with 20 healthy children which had taken physical examination. PBMCs were isolated from peripheral blood and divided into four groups：the blank group, phorbol 12-myristate-13-acetate group （ PMA）, PMA ＋ ASA（5 mmol/L） ,PMA ＋ Tan Ⅱ A （ 10 mg/L）. Each group was cultured in carbon dioxide incubator for 48 hours. Activation rate of NF- κB in PBMCs was determined by immunocytochemical, and the concentrations of TNF-α, IL-1β, IL-8 in supernatant were dectected by ELISA method. Results The activation of NF-κB and the expression of TNF-α ,IL-1β,IL-8 in four KD groups were significant higher than those in four comparative control groups （ P = 0. 000, P = 0. 000, P = 0. 000, P = 0. 000）. In KD PMA group, the activation of NF-κB and the expression of TNF-α, IL-1β, IL-8 was significantly higher than those in control PMA group （P 〈0.01 ,P 〈0.01 ,P 〈0.01 ,P 〈0.01 ）;In KD PMA ＋ ASP group and KD PMA ＋ Tan A group, the levels of NF- κB and all cytokines were significantly lower than those in KD PMA group （ P 〈 0.01 ,P 〈 0.01, P 〈 0.01, P 〈 0.01 ）, while in KD PMA ＋ ASP group those were significant lower than those in KD PMA ＋ Tan ⅡA group（P 〈0.01 ,P 〈0.05 ,P 〈0.01 ,P 〈0.01 ）. AII of cytokines concentrations were positively correlated with the activation of NF- κB （ r = 0. 842, P 〈 0.01 ; r = 0. 898, P 〈 0.01 ; r = 0. 801,P 〈 0.01 ）. Conclusion Tan Ⅱ A has an anti-inflammatory effect in peripheral blood mononuclear ceils （PBMCs） of KD ,which might be through the inhibitation of NF-κB activation and decrease the expression of TNF-α, IL-1β, IL-8 ,just as the same as aspirin.

关 键 词：丹参酮ⅡA 阿司匹林 川崎病 核转录因子-ΚB 炎性细胞因子 

分 类 号：R725.9[医药卫生—儿科]