粉尘螨抗原对支气管哮喘患者外周血单个核细胞T-bet和GATA3及FOXP3 mRNA表达水平的影响

Expression of T-bet,GATA-3 and FOXP3 mRNA in asthmatic peripheral blood mononuclear cells stimulated by dermatophagoides farinae antigen

目的对支气管哮喘(简称哮喘)患者外周血单个核细胞(PBMC)经粉尘螨刺激前、后T-bet mRNA、GATA-3 mRNA、FOXP3 mRNA 的表达水平及相互关系进行研究。方法选择25例哮喘患者和15名健康人为研究对象,每例受试者又分为变应原刺激组和未经变应原刺激的自体对照组,采用半定量-逆转录聚合酶链反应(RT-PCR)检测各组受试者 PBMC 培养后 T-bet mRNA、GATA-3mRNA、FOXP3 mRNA 的表达水平。结果哮喘患者 PBMC 中 FOXP3、T-bet 的表达水平分别为0.3±0.4、0.42±0.24,与健康人(0.4±0.3、0.39±0.37)比较差异无统计学意义(t 值分别为0.78、0.38,P均>0.05),哮喘患者 GATA-3(1.0±0.3)的表达与健康人(0.8±0.3)比较差异有统计学意义(t=2.27,P<0.05);健康人 PBMC 经变应原刺激后 FOXP3的表达(0.72±0.61)与未经变应原刺激的自体对照组(0.27±0.21)比较差异有统计学意义(t=3.12,P<0.01);而 T-bet、GATA-3的表达分别为0.3±0.4、0.9±1.0,与未经变应原刺激的自体对照组(0.3±0.6、0.8±0.6)比较差异无统计学意义(t 值分别为0.27、0.77,P 均>0.05);哮喘患者 PBMC 经变应原刺激后 T-bet、GATA-3的表达分别为0.49±0.42、2.1±1.7,与未经变应原刺激的自体对照组(0.16±0.06、0.6±0.3)比较差异有统计学意义(t 值分别为3.72、5.15,P 均<0.01);但哮喘患者 PBMC 中经变应原刺激后 FOXP3的表达为0.5±0.4,与未经变应原刺激的自体对照组(0.4±0.4)比较差异无统计学意义(t=1.60,P>0.05);经变应原刺激后哮喘患者 PBMC 中 AT-bet、AGATA-3、AFOXP3的表达分别为0.33±0.39、1.58±1.44、0.11±0.32,与健康人(0.03±0.40、0.11±0.53、0.43±0.66)比较差异有统计学意义(t 值分别为2.30、3.79、2.08,P 分别<0.05、<0.01、<0.05);哮喘患者ΔFOXP3与ΔGATA-3、ΔT-bet 表达水平呈负相关(r 值分别为-0.62、-0.46,P 分别<0.01、<0.05)。结论哮喘患者经变应原刺激后,既存在 Th2特定转录因子 GATA-3的优势表达,也存在 Th1特定转录因子 T-bet 的优势表达,同时还存在调节性 T 淋巴细胞(Treg)特定转录因子 FOXP3的表达低下。

Objective To study the expression of T-bet, GATA-3 and FOXP3 mRNA in asthmatic peripheral blood mononuclear cells （PBMC） before and after stimulation by Dermatophagoides farinae antigen （Df） , and the relationship among the three transcription factors. Methods Twenty-five patients of asthma and 15 healthy controls were included. The mRNA levels of T-bet, GATA-3, and FOXP3 in PBMC before and after Df stimulation were detected by semiquantitative reverse transcription-polymerase chair reaction （RT-PCR）. Results There was no statistical difference （ t = 0.78,0.38 respectively, all P 〉 0. 05） in the levels of T-bet and FOXP3 mRNA between the patients（0. 3±0. 4,0.42±0. 24 respectively） and the healthy controls（0.4 ±0. 3,0.39±0.37 respectively）. However, GATA-3 mRNA was statistically different （ t = 2.27, P 〈 0.05 ） between the patients （ 1.0 ±0.3 ） and the healthy controls （ 0.8±0.3）. After the stimulation of PBMC by Df, the levels of T-bet, GATA-3 and FOXP3 mRNA showed statistical difference （ t = 2.30, 3.79,2.08 respectively, P 〈 0.05, 〈 0.01, 〈 0.05 respectively） between the patients （0.33±0. 39, 1.58 ±1.44,0.11±0.32 respectively） and the healthy controls （0.03±0.40, 0.11±0.53,0.43±0.66 respectively）. The FOXP3 mRNA level in the asthmatics was negatively correlated with T-bet and GATA-3 mRNA （r= -0.46,P〈0.05,r= -0.62,P〈0.01 respectively）. Conclusions There are a predominant expression of GATA-3 and T-bet, but a decreased expression of FOXF3 in Dr-stimulated PBMCs from patients with asthma.