KD患儿血清对细胞活力及NF-κB表达的影响和PDTC的干预  

Effects of Serum from KD Patients on HUVEC viability and Expression of NF-κB and the Intervention of PDTC

作　　者：薛超超 李丰[1] 仇慧仙[1] 陈其[1] 张园海[1] 

机构地区：[1]温州医科大学附属育英儿童医院,325027

出　　处：《医学研究杂志》2015年第7期83-87,共5页Journal of Medical Research

基　　金：浙江省医药卫生科技计划项目(201478482)

摘　　要：目的研究川崎病(KD)患儿血清对人脐静脉内皮细胞(HUVEC)细胞活力及核转录因子-κB(NF-κB)表达的影响,探讨KD及其并发症的发病机制。观察吡咯烷二硫氨基甲酸酯(PDTC)干预下HUVEC细胞活力及NF-κB表达的变化,探讨PDTC的作用机制和潜在的临床应用价值。方法根据不同的干预方法将培养的细胞分成4组:N组:RPMI1640培养基+正常儿童血清,K组:RPMI1640培养基+KD患儿血清,G组:RPMI1640培养基+KD患儿血清+IVIG;P组:RPMI1640培养基+KD患儿血清+PDTC。细胞培养24h后,MTT法检测细胞活力,RT-PCR法检测细胞内NF-κB p65m RNA的表达,Western blot法检测细胞核内NF-κB p65蛋白的合成。结果 K组HUVEC增殖受限,细胞活力较N组显著下降(P<0.05),NF-κB p65m RNA、NF-κB p65蛋白表达较N组显著升高(P<0.05);G组与P组细胞活力较K组显著增强(P<0.05),NF-κB p65m RNA、NF-κB p65蛋白表达均较K组显著降低。结论 KD患儿血清能促使细胞内NF-κB信号通路过度激活,导致细胞自身损伤,推测与KD的发生、发展密切相关。PDTC能有效抑制细胞内NF-κB信号通路,对细胞起保护作用,在KD的治疗上有潜在的临床价值。Objective To investigate the influence on the viability of human umbilical vein endothelial cell （ HUVEC） and its expression of NF-κB stimulated by serum from Kawasaki disease （ KD） patients, and discuss the possible pathogenesis of KD and its com-plications.To observe the intervention effect of Pyrrolidine dithiocarbamate （ PDTC） on the cell viability and the expression of NF-κB to explore its mechanism and potential clinical value.Methods HUVECs were divided into 4 groups,and each group was given different in-terfere respectively as following：Group N （ RPMI-1640 medium＋normal serum ）; Group K （ RPMI -1640 medium ＋KD serum ）;Group G（ RPMI-1640 medium＋KD serum＋IVIG）;Group P（ RPMI-1640 medium＋KD serum＋PDTC） .The cells in each group were cultured for 24 hours before cell viabilities were tested by MTT.The expressions of NF-κB p65mRNA were measured by RT-PCR and the expressions of the NF-κB p65 protein by Western blot.Results The cell growth in Group K was inhibited,and its viability was significantly lower than that in Group N （P〈0.05）.The expressions of NF-κB p65mRNA and NF-κB p65 protein in Group K were both significantly higher than that in Group N （P〈0.05）.The viabilities of cells in Group G and P both increased significantly than those in Group K （P〈0.05）.The expressions of NF-κB p65mRNA and NF-κB p65 protein in Group G and P were all much lower than those in Group K （P〈0.05）.Conclusion KD serum can make NF-κB signaling pathway excessive express and lead to cell-self le-sion, which may play an important role on the pathogenesis of KD.PDTC can effectively inhibit NF-κB signaling pathway excessive ex-pression and protect the cell, which indicates it has potential clinical value for KD.

关 键 词：川崎病 核因子 人脐静脉内皮细胞 吡咯烷二硫氨基甲酸酯 

分 类 号：R72[医药卫生—儿科]