川崎病患者外周血淋巴细胞凋亡降低及其机理的研究

Study on the Mechanism of Inhibited Peripheral Blood Lymphocyte Apoptosis in Children with Acute Kawasaki Disease

为探讨外周血单个核细胞（ＰＢＭＣ）凋亡在川病（ＫＤ）发病中的作用。以正常儿童ＰＢＭＣ为对照，观察ＫＤ患者ＰＢＭＣ体外经ａｎｔｉ－ＣＤ３刺激培养后的凋亡细胞百分率和片断ＤＮＡ分析；测定ＰＢＭＣ体外ＰＨＡ导培养产生ＩＬ－６水平，并利用流式细胞你和斑点杂交检测外周血淋巴细胞Ｐ＾５３基因表达水平。与正常对照比较，ＫＤ患者ＰＤＭＣ体外培养产生ＩＬ－６水平明显增高，凋亡细胞百分率降低，片断ＤＮＡ出现时间延迟，

To observe the role of peripheral blood lymphocyte apoptosis in the pathogenesis of Kawasaki disease (KD). Calculating percentage of apoptotic cells and assaying DNA fragmentation in peripheral blood mononuclear cell (PBMC)stimulated by anti-CD3 mAb for oh, 12h, 24h, 48h, 72h in vitro from 30 KD patients and 20 age-matched healthy children,measuring in vitro production of IL -- 6 by ELISA, Determining the expression of PDMC p53 gene mRNA and P53 protein using dot--blot and flow cytometry(FCM) were carried out. The results revealed that apoptotic cell percentage was markedly decreased (P < 0.001 ) and DNA fragmentation delayed,compared with those in normal controls, which could reach the states of control group only by 72h culture,Remarkedely increased production of IL--6 in cultrued supernatant of PBMC induced by phytohemagglutinin (PEA) from KD patients was found,and the expression of PBMC p53 gene mRNA and P53 protein was decreased in KD patients. Adding antiIL-6mAb to the culture led to significantly decreased production of IL-6 (P <0. 001). and reversed decreased apoptotic cell percentage and delayed DNA fragmentation,and the expression of PBMC p53 gene mRNA and P53 protein increased. The data suggested that peripheral blood lymphocyte apoptosis was delayed in KD patient. It might be related to over-production of IL-6 which inhibites the expression of p53 gene.