川崎病白细胞介素-4基因组蛋白乙酰化修饰改变及意义  

Modification and significance of histone acetylation associated with interleukin-4 gene in pediatric Kawasaki disease

作　　者：吕元红[1] 王国兵[1] 温鹏强[1] 刘琮[2] 徐明国[2] 梅洁花 赖恒 李成荣[1] Lyu Yuanhong;Wang Guobing;Wen Pengqiang;Liu Cong;Xu Mingguo;Mei Jiehua;Lai Heng;Li Chengrong(Shenzhen Institute of Pediatrics,Shenzhen Children′s Hospital,Shenzhen 518038,Guangdong Province,China;Department of Cardiology,Shenzhen Children′s Hospital,Shenzhen 518038,Guangdong Province,China)

机构地区：[1]深圳市儿童医院儿科研究所,广东深圳518038 [2]深圳市儿童医院心血管内科,广东深圳518038

出　　处：《中华实用儿科临床杂志》2020年第6期462-466,共5页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金(81102227);深圳市知识创新计划(JCYJ20180228175700233);深圳市卫生计生系统科研项目(201401053)。

摘　　要：目的探讨白细胞介素-4(IL-4)基因组蛋白乙酰化修饰水平改变及其在川崎病(KD)发病机制中的作用。方法选取2016年10月至2018年12月在深圳市儿童医院就诊的KD患儿36例为研究对象,同年龄健康儿童28例作为对照组。KD患儿分别于急性期及静脉用丙种球蛋白(IVIG)治疗有效后4~5 d取血备检。采用染色质免疫共沉淀-荧光定量PCR检测外周血CD4+T淋巴细胞IL-4基因启动子、增强子Va组蛋白H4乙酰化和p300、CREB结合蛋白(CBP)水平;流式细胞术检测外周血Ⅱ型辅助性T淋巴细胞(Th2)(CD4+IL-4+)比例及CD4+T淋巴细胞中磷酸化信号转导及转录活化因子6(pSTAT6)、GATA结合蛋白3(GATA3)、活化T细胞核因子1(NFAT1)、Ⅱ型转化生长因子β受体(TGF-βRⅡ)、磷酸化L型氨基酸转运蛋白1(pLAT1)蛋白表达水平;荧光定量PCR检测CD4+T淋巴细胞IL-4、IL-5、IL-13、IL-4受体α(IL-4Rα)、Ⅰ型转化生长因子β受体(TGF-βRⅠ)、性别决定区Y框蛋白4(SOX4)mRNA表达水平;酶联免疫吸附试验测定血浆IL-4、转化生长因子β(TGF-β)水平。结果1.KD患儿Th2细胞比例、功能相关分子(IL-4、IL-5和IL-13)表达及IL-4基因启动子、增强子Va组蛋白乙酰化水平均明显高于对照组,差异均有统计学意义(均P<0.05),其中冠状动脉损伤组(CAL)前述指标均高于无冠状动脉损伤组(NCAL),差异均有统计学意义(均P<0.05),经IVIG治疗后显著降低,差异均有统计学意义(均P<0.05)。2.与对照组比较KD患儿外周血CD4+T淋巴细胞p300、CBP与IL-4基因启动子、增强子Va结合水平明显上调,差异均有统计学意义(均P<0.05),且p300与IL-4基因启动子、增强子Va结合水平与后者表达均呈正相关(r=0.72、0.43,均P<0.05),经IVIG治疗后呈不同程度下降,差异均有统计学意义(均P<0.05)。其中CAL组p300、CBP与IL-4基因启动子、增强子Va结合水平明显高于NCAL组,差异均有统计学意义(均P<0.05)。3.与对照组比较,KD患儿血浆IL-4水平及Objective To investigate the histone acetylation of interleukin-4(IL-4)gene and its roles in immunological pathogenesis of Kawasaki disease(KD).Methods Thirty-six children with KD and 28 age-matched healthy children in Shenzhen Children′s Hospital from October 2016 to December 2018 were recruited in this study.Peripheral venous blood samples were collected from healthy controls(28 cases)and patients with KD during acute phase and 4 to 5 days after effective intravenous immunoglobulin(IVIG)treatment.Co-immunoprecipitation followed by real-time PCR was used to assess histone H4 acetylation levels of IL-4 promoter and Va enhancer,and binding abilities of p300 and CREB-binding protein(CBP)with promoter and Va enhancer of IL-4 gene in peripheral blood CD4+T cells.Flow cytometry was performed to analyze the proportion of CD4+IL-4+T cells(Th2)and protein le-vels of phosphorylated signal transducer and activator of transcription 6(pSTAT6),GATA binding protein 3(GATA3),nuclear factor 1 of activated T cells(NFAT1),transforming growth factor-βreceptorⅡ(TGF-βRⅡ),and phosphorylated L-type amino acid transporter 1(pLAT1).Quantitative real-time PCR was used to evaluate the transcription levels of IL-4,IL-5,IL-13,IL-4 receptorα(IL-4Rα),transforming growth factor-βreceptorⅠ(TGF-βRⅠ)and sex-determining region Y(SRY)-box 4(SOX4)in CD4+T cells.Plasma concentrations of IL-4 and transforming growth factor-β(TGF-β)were measured by enzyme-linked immunosorbent assay.Results(1)Compared with control group,the proportion of Th2 cells,expression levels of Th2-associated cytokines(IL-4,IL-5 and IL-13)and histone H4 acetylation levels associating with IL-4 promoter and Va enhancer,increased remarkably during acute KD(all P<0.05),and restored after IVIG therapy(all P<0.05).Meanwhile,all the former items in KD patients with coronary artery lesions(CAL)were higher than those in patients with non-coronary artery lesions(NCAL)(all P<0.05).(2)Compared with control group,binding abilities of p300 and CBP with IL-4 promoter and Va en

关 键 词：川崎病 Ⅱ型辅助性T淋巴细胞 细胞因子 组蛋白乙酰化 免疫 

分 类 号：R72[医药卫生—儿科]