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目的:观察T细胞特异转录因子T-bet/GATA-3在支气管哮喘中失衡表达,探讨黄芪对其调节作用。方法:50只雄性SPF级SD大鼠随机分为对照组(C组)、哮喘组(A组)、黄芪低(H1组)、中(H2组)、高(H3组)剂量组,20只雄性SPF级致敏SD大鼠脾脏中分离获得CD4+T细胞,分为对照组(C组),OVA干预组(A组),黄芪干预组低(H1组)、中(H2组)、高(H3组)。HE染色观察气道病理改变;酶联免疫吸附测定(ELISA)法检测细胞因子(IL)-4,5和IFN-γ含量;逆转录-聚合酶链反应(RT-PCR)测定法检测T-bet和GATA-3mRNA表达;免疫印迹(Westernblot)法检测T-bet和GATA-3蛋白表达。结果:在大鼠肺组织中嗜酸粒细胞(EOS)、淋巴细胞、管壁面积/支气管管腔内周长(WA/Pi)和支气管平滑肌面积/支气管管腔内周长(ASM/Pi)A组比C组明显增多或增厚(P均<0.01),H1、H2、H3组中不同程度的减少或减轻(P>0.05);血清中IL-4和IL-5含量A组高于C组(P均<0.01),H1、H2、H3组剂量组均低于A组(P均<0.01),血清中IFN-γ含量A组远低于C组(P均<0.01),H1、H2、H3组明显高于A组(P均<0.01),CD4+T细胞培养上清液中H1、H2、H3组IL-4和IL-5含量均降低,同时增加IFN-γ含量,与A组比较,差异均有统计学意义(P均<0.01);肺组织和CD4+T细胞中T-betmRNA和蛋白表达H1、H2、H3组高于A组(P均<0.01);而GATA-3mRNA和蛋白表达H1、H2、H3组低于A组(P均<0.01)。结论:T细胞特异转录因子T-bet/GATA-3在哮喘中失衡表达,黄芪抑制哮喘气道炎症可能通过双向调节T-bet和GATA-3平衡来实现。
OBJECTIVE: To observe the imbalanced expression of T-cell transcription factor T-bet / GATA-3 in bronchial asthma and to investigate the regulation effect of astragalus. Methods: Fifty male SPF SD rats were randomly divided into control group (C group), asthma group (A group), astragalus low (H1 group), medium (H2 group) and high (H3 group) CD4 + T cells were isolated from spleen of male SPF-sensitized SD rats and divided into control group (group C), OVA intervention group (group A), astragalus intervention group low (group H1), medium (group H2) (Group H3). The pathological changes of airway were observed by HE staining; the contents of cytokines (IL) -4 and 5 and IFN-γ were detected by enzyme linked immunosorbent assay (ELISA) bet and GATA-3mRNA expression. The expression of T-bet and GATA-3 protein were detected by Western blotting. RESULTS: Eosinophils (EOS), lymphocytes, wall area / bronchial lumen perimeter (WA / Pi) and bronchial smooth muscle area / bronchial lumenal perimeter (ASM / Pi) The levels of IL-4 and IL-5 in serum in group A were significantly higher than those in group C (P <0.01), but were decreased in groups of H1, H2 and H3 The levels of IFN-γ in serum of group A and group C were significantly lower than those of group C (all P <0.01), and the levels of H1, H2 and H3 were lower than those of group A (all P <0.01) (P <0.01). The contents of IL-4 and IL-5 in H1, H2 and H3 in the culture supernatant of CD4 + T cells were decreased, while the content of IFN- Compared with group A, the difference was statistically significant (all P <0.01). The expression of T-bet mRNA and protein in lung tissue and CD4 + T cells were higher than those in group A (all P <0.01) The expression of GATA-3mRNA and protein in H1, H2 and H3 groups were lower than those in A group (all P <0.01). CONCLUSION: T-cell-specific transcription factor T-bet / GATA-3 is imbalanced in asthma. Astragalus may play an important role in airway inflammation in asthma by bidirectional regulation of T-bet and GATA-3 balance.