T淋巴细胞免疫球蛋白黏蛋白-3mRNA表达在支气管哮喘中的作用  

Expression of T cell immunogloblin and mucin domain-containing molecule 3 mRNA and its relationship with asthma

作　　者：韩锋[1] 陆小霞[1] 王莹[1] 

机构地区：[1]武汉市妇女儿童医疗保健中心呼吸内科,430016

出　　处：《中华实用儿科临床杂志》2014年第21期1614-1617,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金青年科学基金（81200006）;吴阶平医学基金（320.6750.1285）;湖北省自然科学基金（2010CDB08804）;武汉市卫生局科研项目（WX11B10）

摘　　要：目的观察支气管哮喘（哮喘）患儿和哮喘小鼠外周血单个核细胞（PBMC）中T淋巴细胞免疫球蛋白黏蛋白-3（Tim-3）mRNA的表达，探讨Tim-3在哮喘中的作用。方法选取2012年1月至2013年6月于武汉市妇女儿童医疗保健中心呼吸内科住院或哮喘门诊患儿97例，哮喘发作期47例（哮喘发作组），哮喘缓解期50例（哮喘缓解组），健康体检儿童42例为健康对照组。检测3组儿童PBMC中Tim-3mRNA的表达，采用酶联免疫吸附试验（ELISA）法检测血浆中白细胞介素-6（IL-6）、转化生长因-β（TGF-β）的水平，并分析Tim-3mRNA与IL-6水平的相关性，同时建立小鼠哮喘动物模型，分为正常对照组和哮喘组，每组10只，应用实时荧光定量PCR检测小鼠PBMC中Tim-3mRNA的水平，流式细胞分析技术检测CIM细胞中IL-17细胞比例反映Th17的水平，CD4细胞中CD25^＋FoxP3^＋反映调节性T淋巴细胞（Treg）的水平；并分析Tim-3的表达与Th17／Treg水平的相关性。结果儿童哮喘发作组PBMC中Tim-3mRNA吸光度值高于哮喘缓解组和健康对照组（P均〈0．05），儿童哮喘发作组血浆中IL-6水平高于哮喘缓解组和健康对照组（P均〈0．05），动物模型中哮喘组PBMC中Tim-3mRNA的水平高于正常对照组（P〈0．01），哮喘组CD4^＋IL-17^＋表达比例高于正常对照组，而CD4^＋CD25^＋FoxP3^＋低于正常对照组（P均〈0．05），哮喘组Th17／Treg的比值高于正常对照组（P〈0．05），哮喘组PBMC中Tim-3mRNA的表达水平与CD4^＋IL-17^＋及Th17／Treg的比例呈正相关，与CD4^＋CD25^＋FoxP^3＋的水平呈负相关。结论哮喘患儿PBMC中Tim-3mRNA表达增高，参与IL-6相关的呼吸道炎性反应。动物实验提示Tim-3与Th17／Treg失衡密切相关，表明Tim-3可能在哮喘的发生、发展中起着一定的作用。Objective To detect the expression of T cell immunogloblin and mucin domain-containing male- cule 3 （Tim-3） mRNA in peripheral blood mononuclear cells （PBMC） isolated from asthmatic children and asthmatic murine,and to explore the role of Tim-3 in bronchial asthma. Methods Recruiting 97 cases asthmatic children,47 children with acute asthma exacerbation（ acute attack group） ,50 children with asthma remission（ remittent group）and 42 children as the healthy control group. The Tim-3 mRNA expression was measured by using reverse transcription-pol- ymerase chain reaction （RT-PCR） method. The levels of interleukin-6 （IL-6） and transforming growth factor-~ （TGF-[3） were determined with enzyme-linked immunosorbent assay （ELISA）, and to analyze the correlation between Tim-3 mRNA and the level of IL-6. The asthmatic murine models were randomly divided into 2 groups ： normal control group and asthma group. There were 10 mice in each group. PBMC in each group were collected. Expression of Tim-3 mRNA was detected by using real-time PCR. The level of CD4 ^＋ IL-17 ^＋ and CD4 ^＋ CD25^＋ FoxP3 ^＋ which reflected Thl7 and regulatory T cells（Treg） expression respectively were detected by using flow cytometry, and the correlation between Tim-3 and Thl7/Treg level was analyzed. Results Expression of Tim-3 mRNA in children with acute asthma exacerbation was significantly higher than those in remission stage and healthy control group （ all P 〈 0.05 ）. The expression of Tim-3 mRNA in children with asthma remission was significantly higher than that in healthy control group （ P 〈 0.05 ）, and the level of IL-6 in children with acute asthma exacerbation was significantly higher than those in remission stage and healthy control group （ all P 〈 0.05 ）. In the murine model of asthma group, the level of Tim-3 mRNA in PBMC was significantly higher than that in normal control group （ P 〈 0.01 ）, and in asthma group CD4^＋ IL-17 ^＋ expression was significantly higher than

关 键 词：支气管哮喘 T细胞免疫球蛋白黏蛋白-3 MRNA T淋巴细胞 

分 类 号：R725.6[医药卫生—儿科]