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目的研究布地奈德(BUD)对支气管哮喘大鼠支气管信号转导子和转录激活子6(STAT6)基因和蛋白表达的调控作用。方法30只清洁级幼年雄性SD大鼠随机分为对照组(A组)、哮喘组(B组)和BUD组。对支气管肺泡灌洗液(BALF)进行细胞总数、嗜酸性粒细胞(EOS)计数和分类计数;应用双抗体夹心酶联免疫吸附试验法测定BALF中IL-4、IL-12浓度;采用免疫组化法和原位杂交法分别检测STAT6蛋白和STAT6mRNA表达的变化。结果(1)B组BALF中细胞总数、EOS绝对值和EOS占细胞总数的百分比(EOS%)均显著高于A组(P<0.01),BUD组BALF中上述各项指标较B组均显著降低(P<0.01);(2)BALF中IL-4的浓度B组显著高于A组(P<0.01),BUD组较B组显著降低(P<0.01),而IL-12的浓度B组显著低于A组(P<0.01),BUD组较B组显著升高(P<0.01);(3)B组支气管上皮细胞STAT6蛋白和STAT6mRNA阳性表达较A组明显增强(均为P<0.01),BUD组较B组明显减弱(均为P<0.01);(4)支气管上皮细胞STAT6蛋白、STAT6mRNA分别与BALF中的IL-4浓度呈显著正相关,与BALF中EOS绝对值呈显著正相关;而与IL-12浓度呈显著负相关。结论哮喘大鼠支气管STAT6及其mRNA较强表达,上皮细胞是其主要表达细胞;BUD有抑制气道炎症的作用,下调STAT6及其基因表达,使IL-4合成减少可能为其重要作用机制。
Objective To investigate the regulatory effect of budesonide (BUD) on the gene and protein expression of bronchial astaxanthin and activator of transcription 6 (STAT6) in bronchial asthmatic rats. Methods Thirty clean male SD rats were randomly divided into control group (group A), asthma group (group B) and BUD group. The numbers of total cells, eosinophils (EOS) were counted and classified in bronchoalveolar lavage fluid (BALF). The concentrations of IL-4 and IL-12 in BALF were measured by double antibody sandwich enzyme-linked immunosorbent assay The changes of STAT6 protein and STAT6 mRNA expression were detected by hybridization and in situ hybridization. Results (1) The percentage of total cell number, absolute EOS and total cell percentage of EOS in BALF of group B were significantly higher than that of group A (P <0.01). The above indexes of BALF in group BUD were significantly higher than that of group B (P <0.01). (2) The concentration of IL-4 in BALF in group B was significantly higher than that in group A (P <0.01), and the level of IL-12 in group BUD was significantly lower than that in group B (P <0.01). (3) The positive expression of STAT6 and STAT6 in bronchial epithelial cells in group B were significantly higher than those in group A (all P < (P <0.01). (4) The STAT6 and STAT6 mRNA in bronchial epithelial cells were positively correlated with the concentration of IL-4 in BALF and the absolute value of EOS in BALF But negatively correlated with IL-12 concentration. Conclusions The expression of STAT6 and its mRNA in asthmatic rats is stronger than that in epithelial cells. BUD may play an important role in inhibiting airway inflammation, down-regulating STAT6 and its gene expression and decreasing IL-4 synthesis.