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目的 :探讨哮喘小鼠模型中白细胞介素(interleukin,IL)-25的表达,研究其对肥大细胞IL-6分泌的影响。方法 :6~8周龄BALB/c小鼠随机分为对照组和哮喘组,分别用等量磷酸盐缓冲液(phosphate buffer solution,PBS)或卵清蛋白(ovalbumin,OVA)致敏,检测肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中细胞总数及嗜酸性粒细胞(eosnophils,EOS)计数验证模型的成功建立。两组小鼠肺组织中IL-25 m RNA、BALF及血清中IL-25的表达分别应用逆转录-聚合酶链反应法、酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)检测。小鼠肥大细胞P815分为对照组和IL-25组,分别予等量培养基和IL-25干预,收集细胞培养上清,用ELISA检测IL-6的水平。结果:哮喘组BALF中细胞总数及EOS计数较对照组显著升高(P<0.01),小鼠出现呼吸频率加快、烦躁不安等表现,模型建立成功。哮喘组肺组织中IL-25 m RNA、BALF及血清中IL-25表达较正常组显著升高(P<0.01)。细胞水平的研究发现IL-25组P815上清中IL-6表达较对照组显著升高(P<0.01)。结论:IL-25在OVA诱导哮喘小鼠模型中表达升高。IL-25刺激肥大细胞分泌IL-6,可能在哮喘的发病中起促进作用。
Objective: To investigate the expression of interleukin-25 (IL-25) in asthmatic mouse model and its effect on the secretion of IL-6 in mast cells. Methods: BALB / c mice (6-8 weeks old) were randomly divided into control group and asthma group, sensitized with phosphate buffered saline (PBS) or ovalbumin (OVA) The total number of cells in the bronchoalveolar lavage fluid (BALF) and eosinophils (EOS) count validation model was successfully established. The expression of IL-25 m RNA, BALF and serum IL-25 in lung tissue of the two groups were detected by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA) respectively. Mice mast cells P815 were divided into control group and IL-25 group, respectively, with equal medium and IL-25 intervention, cell culture supernatants were collected, and the level of IL-6 was detected by ELISA. Results: The total number of cells in BALF and the count of EOS in asthma group were significantly higher than those in control group (P <0.01). The mice showed respiratory rate and irritability. The model was established successfully. The levels of IL-25mRNA, IL-25 in BALF and serum in asthma group were significantly higher than those in normal group (P <0.01). The level of IL-25 in the supernatant of P815 group was significantly higher than that in the control group (P <0.01). Conclusion: IL-25 is up-regulated in OVA-induced asthma mouse model. IL-25 stimulates mast cells to secrete IL-6, which may play a role in the pathogenesis of asthma.