摘要
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 T cell-specific transcription factors T-bet and GATA-3 in asthmatic rats and discuss the regulation of the Astragalus membranaceus acts on them.Methods：Fifty male SD rats were randomly divided into Group C（control group）,Group A（asthmatic group）,Group H1（low-dose Astragalus membranaceus group）,Group H2（medium-dose Astragalus membranaceus group） and Group H3（high-dose Astragalus membranaceus group）,which were observed the changes of airway histology.CD4＋T cells were isolated from twenty male SD rats spleens and incubated with RPMI-1640 at 37℃ for 24 hours,and were divided into 5 groups as noted above.The concentrations of interleukin（IL）-4,IL-5,IFN-γ in serum and CD4＋T cells supernatant were measured by enzyme-linked immunosorbent assay（ELISA）,while the mRNA expressions of T-bet and GATA-3 were detected by reverse transcription-polymerase chain reaction（RT-PCR）,and the protein expressions of T-bet and GATA-3 were detected by western blot respectively.Results：The numbers of eosinophils（EOS） and lymphocytes,the thicknesses of WA /Pi and ASM /Pi of Group A,were significantly elevated compared with those of Group C（P 〈 0.01）.The EOS and lymphocytes,the thicknesses of WA/Pi and ASM/Pi of Group H1-3 were lower than those of Group A.The IL-4 and IL-5 concentrations of Group A were significantly elevated compared with those of Group C,and those of Group H1-H3 were lower than Group A（P 〈 0.01）.Compared with Group C（P 〈 0.01）,the significantly lower concentration of IFN-γ in Group A was detected,and the Group H1-3 had the higher concentrantions of IFN-γ than Group A（P 〈 0.01）.In CD4＋T cells supernatant,the concentrations of IL-4 and IL-5 were distinctly decreased and the concentrantions of IFN-γ were increased in the Group H1-3.There were obvious differences in the concentrations of IL-4,IL-5 and IFN-γ between the Group H1-3 and Group A（P 〈 0.01）.In Group H1-3,the expressions of T-bet mRNA and protein in lungs and CD4＋T cells were obviously increased compared with those of Group A（P 〈 0.01）;and the Group H1-3 has the lower levels of GATA-3 mRNA and protein expressions than Group A（P 〈 0.01）.Conclusion：In asthma the imbalanced T cell-specific transcription factors T-bet and GATA-3 play a key role in Th2 polarization with over-production of Th2-like cytokines.Astragalus membranaceus inhibits the asthmatic airway inflammation with regulating the balance of transcription factors Tbet and GATA-3.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2010年第9期1249-1254,共6页
Journal of Nanjing Medical University(Natural Sciences)
基金
无锡市科技局项目(CSZ200716)
关键词
哮喘
转录因子
T淋巴细胞
黄芪
asthma； transcription factors； T-lymphocyte； Astragalus membranaceus
