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目的探讨转录因子T-bet和GATA-3对小儿喘息性支气管炎的免疫调控作用。方法对32例喘息性支气管炎(喘支组)和30例正常儿童(对照组)抽取抗凝静脉血5 mL,Ficoll分离外周血淋巴细胞,以终质量浓度为100 mg/L的植物血凝素(PHA)刺激48 h,采用酶联免疫吸附法(ELISA)检测培养上清液干扰素-γ(IFN-γ)和白细胞介素-4(IL-4)水平,用逆转录聚合酶链式反应(RT-PCR)检测淋巴细胞T-bet、GATA-3的mRNA表达水平。结果喘支组淋巴细胞培养上清液IFN-γ水平[(528±164)ng/L]低于对照组[(870±203)ng/L](P<0.05),而IL-4水平[(106±29)ng/L]明显高于对照组[(47±15)ng/L](P<0.01);喘支组和对照组淋巴细胞T-bet mRNA水平分别为(0.11±0.03)(、0.17±0.03),GATA-3 mRNA分别为(0.54±0.09)、(0.41±0.07),提示喘支组T-bet mRNA表达较对照组减少(P<0.05),而GATA-3 mRNA明显高于对照组(P<0.01);喘支组IFN-γ水平与T-bet mRNA表达I、L-4与GATA-3 mRNA表达均呈正相关(r=0.57,0.60 P均<0.01);IFN-γ与IL-4水平、T-bet mRNA与GATA-3 mRNA表达均呈负相关(r=-0.38,-0.46 P均<0.05)。结论1.喘支组患儿淋巴细胞合成IL-4水平增高,IFN-γ减少,存在以Th2细胞过度分化为特征的T辅助细胞分化失衡。2.喘支组患儿T细胞分化失衡受到核转录因子T-bet和GATA-3的调控,T-bet表达减少可能是一重要因素。
Objective To investigate the immunomodulatory effects of transcription factors T-bet and GATA-3 on asthmatic bronchitis in children. Methods 32 mL of asthmatic bronchitis (asthma group) and 30 normal children (control group) were drawn anticoagulated venous blood 5 mL. Peripheral blood lymphocytes were isolated by Ficoll, and the final concentration of 100 mg / L phytohemagglutination (IFN-γ) and interleukin-4 (IL-4) in culture supernatants were detected by enzyme-linked immunosorbent assay (ELISA) The mRNA expression of T-bet and GATA-3 in lymphocytes was detected by RT-PCR. Results Compared with the control group (870 ± 203 ng / L), the level of IFN-γ in the supernatant of the bronchial asthma group (528 ± 164 ng / L) was significantly lower than that of the control group (P <0.05) (106 ± 29) ng / L] were significantly higher than those in the control group [(47 ± 15) ng / L] (P 0. 01). The levels of T-bet mRNA in the asthmatic group and the control group were (0.11 ± 0.03) (0.17 ± 0.03) and GATA-3 mRNA (0.54 ± 0.09) and (0.41 ± 0.07), respectively, suggesting that the expression of T-bet mRNA in asthmatic group was significantly lower than that in control group (P <0.05) (P <0.01). The level of IFN-γ in the asthma group was positively correlated with the expression of T-bet mRNA, the mRNA expression of L-4 and GATA-3 in the asthmatic group (r = 0.57,0.60 P <0.01) -γ and IL-4 levels. There was a negative correlation between T-bet mRNA and GATA-3 mRNA expression (r = -0.38, -0.46 P <0.05). In the asthmatic group, the level of IL-4 in the lymphocytes was increased and the IFN-γ was decreased. There was imbalanced T helper cell differentiation characterized by Th2 cell hyper-differentiation. The imbalance of T cells in children with asthma was regulated by T-bet and GATA-3, and the decrease of T-bet might be an important factor.