地塞米松对哮喘大鼠T-bet?GATA-3及气道炎症的作用机制
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江苏省自然科学基金资助(BK2005152)


Mechanism of action of dexamethasone on expression of T-bet?GATA-3 and airway inflammation in asthmatic rats
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    摘要:

    目的:观察地塞米松对哮喘大鼠气道炎症的作用以及对转录因子T-bet-GATA-3表达的影响-方法:36只SPF级SD大鼠随机分为正常对照组(A组)-哮喘模型组(B组)和地塞米松组(C组),每组12只-以卵蛋白(OVA)腹腔注射并雾化吸入制备大鼠哮喘模型,观察3组大鼠气道病理改变;采用ELISA法测定肺泡灌洗液(BALF)中IL-2-IL-4-IL-5和IFN-γ含量;采用RT-PCR和Western blotting检测T-bet和GATA-3 mRNA和蛋白的表达-结果:①予OVA腹腔注射和雾化,哮喘大鼠模型制备成功;②地塞米松减少Th2型细胞因子(IL-4,IL-5)的表达(P < 0.01),对Th1型细胞因子(IL-2,IFN-γ)表达量无明显影响;③地塞米松抑制GATA-3表达(P < 0.01),对T-bet表达无明显影响;④相关分析显示B组T-bet蛋白表达量与EOS-L-WA/Pi和ASM/Pi呈负相关(P < 0.01);B组GATA-3蛋白表达量与EOS-L-WA/Pi和ASM/Pi呈正相关(P < 0.01);B组大鼠T-bet蛋白表达量与GATA-3蛋白表达量呈负相关(P < 0.01)-结论:支气管哮喘大鼠存在T-bet低表达,GATA-3高表达,并且与气道炎症关系密切;地塞米松抑制气道炎症,可抑制GATA-3表达,但对T-bet表达无明显影响-

    Abstract:

    Objective:To observe the effect of dexamethasone on expression of airway inflammation and transcription factor T-bet and GATA-3 in asthmatic rats. Methods:Thirty-six SD rats(SPF) were randomly divided into normal control group(group A),asthmatic model group(group B) and dexamethasone group(group C) with 12 rats in each group. The asthmatic model of SD rats was established by immunization with intraperitoneal injected and inhaled ovalbumin(OVA). The animals were executed under anesthesia within 24 h after the last atomization. The morphological changes in lung tissues of rats were measured. The thickness of airway wall(WA/Pi) and airway smooth muscle(ASM/Pi),and the numbers of eosinophils and lymphocytes were measured. The concentrations of interleukin-2(IL-2),IL-4,IL-5 and interferon-γ(IFN-γ)in bronchoalveolar lavage fluid(BALF) were detected by enzyme-linked immunosorbent assay(ELISA). The mRNA and protein expressions of T-bet and GATA-3 in the lungs were measured semiquantitatively by reverse transcription-polymerase chain reaction(RT-PCR) and Western blotting, respectively. Results:In group B and group C,the numbers of eosinophils and lymphocytes,and the WA/Pi and ASM/Pi were all significantly higher than those in group A(P < 0.01). The four indexes in group C were all significantly lower than those in group B(P < 0.01). In group B and group C,the concentrations of IL-4 and IL-5 in BALF were all significantly higher than those group A(P < 0.01),the concentrations of IL-2 and IFN-γ in BALF were all significantly lower than those in group A(P < 0.01). In group C,the concentrations of IL-4 and IL-5 in BALF were all significantly lower than those group B(P < 0.01),but the concentrations of IL-2 and IFN-γ in BALF were no significantly than those in group B. In group B and group C,expressions of T-bet were significantly lower than that in group A(P < 0.01),but expressions of T-bet in group C were no significantly than that in group B. In group B and group C,expression of GATA-3 were significantly higher than that in group A(P < 0.01). Expression of GATA-3 in group C were significantly lower than that in group B(P < 0.01). The correlation analysis indicated that in group B,protein expressions of T-bet was negatively correlated with the numbers of eosinophils and lymphocytes,and the WA/Pi and ASM/Pi(r = -0.82,-0.86,-0.78,-0.80,P < 0.01),But protein expression of GATA-3 was positively correlated with the numbers of eosinophils and lymphocytes,and the and ASM/Pi(r = 0.83,0.85,0.76,0.79,P < 0.01). Protein expression of T-bet was negatively correlated with protein expression of GATA-3 in group B(r = -0.88,P < 0.01). Conclusion:The expressions of T-bet were low in bronchial asthmatic rats,and the expressions of GATA-3 were high in bronchial asthmatic rats. Dexamethasone can lessen airway inflammation and expression of GATA-3,but there was no significant effect on expression of T-bet with dexamethasone.

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吉宁飞,周玉婷,何 畏,卞 涛,殷凯生.地塞米松对哮喘大鼠T-bet?GATA-3及气道炎症的作用机制[J].南京医科大学学报(自然科学版),2008,28(11):1421-

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