丹参酮ⅡA对川崎病急性期外周血单个核细胞核转录因子-κB活化及炎性细胞因子表达影响研究  

Effect of different concentration of Tanshinone IIA on the activation of nuclear factor-kappa B and the expression of inflammatory cytokines in peripheral blood mononuclear cells (PBMC) of the acute phase of Kawasaki disease

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

机构地区：[1]重庆医科大学附属儿童医院血液科,儿童发育疾病研究省部共建教育部重点实验室,儿科学重庆市重点实验室,重庆市儿童发育重大疾病诊治与预防国际合作科技基地,重庆400014 [2]成都市妇女儿童中心医院血液科(原成都市儿童医院),成都610091

出　　处：《中国实用儿科杂志》2012年第10期757-759,共3页Chinese Journal of Practical Pediatrics

基　　金：四川省卫生厅科研课题[项目编号110061]

摘　　要：目的探讨不同浓度丹参酮ⅡA(TanⅡA)对急性期川崎病(KD)患儿外周血单个核细胞(PBMC)核转录因子-κB(NF-κB)活化及肿瘤坏死因子-α(TNF-α)、白介素-8(IL-8)表达的影响。同时探讨KD发病机制和了解TanⅡA抗炎作用效果。方法研究对象为2010年12月至2011年5月成都市妇女儿童中心医院住院的KD患儿20例(KD组),同期门诊健康体检儿童20例为健康对照组,采用Ficoll密度梯度离心法分离静脉血PBMC,并分组进行培养,即自然培养组(空白)、12-肉豆蔻酰-13-乙酸佛波酯(PMA)、PMA+不同浓度TanⅡA(TanⅡA终浓度分别为1、3、10mg/L),采用免疫细胞化学法观察培养后各组细胞中NF-κB的活化率,通过双抗体酶联免疫吸附法(ELISA)检测各组培养上清液中TNF-α、IL-8的表达量。结果 KD组不同培养条件各指标均高于健康对照组(P<0.05)。加入PMA培养后,KD组和健康对照组各指标均明显高于空白条件,差异有统计学意义(P均<0.05)。加入PMA+3mg/LTanⅡA培养后,健康对照组TNF-α的表达量无明显下降(P>0.05);NF-κB的活化率与TNF-α和IL-8的表达量呈正直线相关关系(r=0.817,P<0.01;r=0.782,P<0.01)。TNF-α与IL-8的表达改变相一致,亦呈正直线相关关系(r=0.709,P<0.01)。结论急性期KD患儿PBMCs中NF-κB活化增强及TNF-α、IL-8表达量增加,参与了急性期KD免疫炎性反应,可能介导了急性期KD免疫血管损伤;TanⅡA在急性期KD患儿PBMCs中的抗炎作用具有剂量依赖性,其抗炎作用机制可能是通过抑制NF-κB的活化,进而抑制其下游TNF-α、IL-8的表达而实现。Objective To explore the inhibition effect of Tanshinone IIA on the activation of nuclear factor-kappa B （NF-κB） and the expression of TNF-α, IL-8 in peripheral blood mononuclear cells （PBMC） of the acute phase of κawasaki disease （κD） and to explore the pathogenesis of κD and to explore the anti-inflammatory effect of Tanshinone IIA. Methods PBMCs were isolated and purified from peripheral blood of 20 κD children and 20 healthy children by density gradient centrifugation. In every sample PBMC were divided into five groups.The first group was cultured naturally, the second one was stimulated by phorbol 12-myristate-13-acetate （PMA）, while other groups were stimulated by PMA and Tanshi-noneIIA which was treated with different concentrations. Each group was cultured in carbon dioxide incubator. Activation of NF-κB in PBMCs was determined by immuno- cytochemistry, and ELISA was used to measure the concentration of TNF-α, IL-8 in supernatant. Results Under PMA culturing, the activation of NF-κB was significantly higher than the blank group both in κD and control group（P〈 0.05） ; in supernatant of the two, the ex-pression of TNF-a, IL-8 was significantly higher than the blank group（P 〈 0.05, P 〈 0.01 ）.Under PMA＋ final concentration of 3mg / 1 Tan II A culturing, the activation of NF-κB in κD and control group was significantly lower than the PMA group （P 〈 0.05 ） ; the expression of IL-8 in κD and control group and the expression of TNF-α in κD group were significantly lower than the PMA group（P 〈 0.05 ）, while the control group, concerning of TNF-α, there was no significant decline （P 〉 0.05）. Under PMA＋ final concentration of 10mg/l Tan II A culturing, the activation of NF-κB in κD and control group was significantly lower than the PMA group （P 〈 0.05 ） ; in supernatant of the κD and control group the expression of TNF-α and IL-8 was significantly lower too, and the PMA＋ final concentration of 10mg/l Tan II A group was lower than the PM

关 键 词：川崎病 核转录因子-ΚB 丹参酮ⅡA 

分 类 号：R72[医药卫生—儿科]