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目的:研究固本防哮饮含药血清对白细胞介素-4(IL-4)刺激的人胚肺成纤维细胞(MRC-5)中解整合素金属蛋白酶(ADAM)33和E26转录因子(ETS)1表达的影响,探讨其可能的抗哮喘作用机制。方法:制备固本防哮饮含药血清,以IL-4刺激MRC-5细胞,采用固本防哮饮含药血清进行干预,24h后收集细胞,Real-time RT-PCR法检测各组细胞中ADAM33、ETS1 mRNA的表达;Western Blot法检测各组细胞中ADAM33、ETS1蛋白的表达;免疫荧光技术检测ADAM33和ETS1在细胞亚结构的定位及相对表达。结果:固本防哮饮含药血清能显著降低IL-4刺激的MRC-5细胞中ADAM33、ETS1 mRNA的高表达(P<0.01),且孟鲁司特钠含药血清对ADAM33 mRNA的下调作用显著优于固本防哮饮组(P<0.01),固本防哮饮含药血清对ETS1 mRNA的下调作用优于孟鲁司特钠组(P<0.05)。固本防哮饮含药血清能降低IL-4刺激的MRC-5细胞中ADAM33蛋白的高表达(P<0.05)和ETS1蛋白的高表达(P<0.01)。ADAM33、ETS1蛋白主要在细胞膜和细胞质中表达,而固本防哮饮含药血清能显著降低IL-4刺激的MRC-5细胞中ADAM33、ETS1平均荧光强度(P<0.01)。结论:固本防哮饮可通过调控ADAM33、ETS1的表达,减轻气道高反应性以及气道重塑,防治支气管哮喘。
Objective: To investigate the effect of Guben anti-asthma drink containing serum on the expression of ADAM33 and E26 transcription factor (IL-4) in human embryonic lung fibroblasts (MRC-5) stimulated by interleukin-4 ETS) 1 expression, explore its possible anti-asthma mechanism. Methods: The drug-containing serum of Guben anti-asthma drink was prepared, MRC-5 cells were stimulated by IL-4, and the drug-containing serum of Guben anti-asthma drink was used for intervention. After 24h, cells were harvested and cells of each group were detected by Real-time RT- The expression of ADAM33 and ETS1 mRNA was detected by Western Blot. The expression of ADAM33 and ETS1 protein was detected by Western Blot. The localization and relative expression of ADAM33 and ETS1 were detected by immunofluorescence. Results: Guben anti-asthma drink-containing serum could significantly reduce the expression of ADAM33 and ETS1 mRNA in IL-4-stimulated MRC-5 cells (P <0.01), and the decrease of ADAM33 mRNA The effect of Guben anti-asthma drink-containing serum on ETS1 mRNA was better than that of montelukast sodium group (P <0.05). Guben anti-asthma drink-containing serum can reduce high expression of ADAM33 protein (P <0.05) and high expression of ETS1 protein (P <0.01) in IL-4-stimulated MRC-5 cells. ADAM33 and ETS1 protein were mainly expressed in the cell membrane and cytoplasm. However, the CCK-containing anti-asthma drink-containing serum could significantly reduce the average fluorescence intensity of ADAM33 and ETS1 (P <0.01) in IL-4-stimulated MRC-5 cells. Conclusion: Guben anti-asthma drink can prevent and treat bronchial asthma by regulating the expression of ADAM33 and ETS1, reducing airway hyperresponsiveness and airway remodeling.