麻黄碱对人支气管上皮细胞IL-8表达的影响

Effect of ephedrine on expression of interleukin-8 in human bronchial epithelial cells

目的观察麻黄碱对人支气管上皮细胞(16HBE)白细胞介素8(IL-8)表达的影响,探讨麻黄治疗哮喘的机制。方法将16HBE随机分为3组,A、B组分别加入20 ng/m L TNF-α、20 ng/m L TNF-α+300μg/m L麻黄碱,C组不处理,培养18 h。收集各组细胞,分别采用Real-time PCR、Western blot法检测IL-8 mRNA及蛋白;收集各组细胞上清液,ELISA法检测IL-8蛋白。结果 A、B组细胞IL-8 mRNA及蛋白表达量均较C组升高,A组较B组IL-8mRNA及蛋白表达量均降低,P均<0.05;A、B组上清液中IL-8蛋白分别为(27.24±1.91)、(23.62±3.40)pg/m L,均高于C组的(17.45±2.32)pg/m L,P均<0.05;A、B组上清液中IL-8蛋白比较,P<0.05。结论麻黄碱能抑制促炎因子TNF-α诱导的16HBE中IL-8的表达及分泌,这可能是麻黄治疗哮喘的机制之一。

Objective To observe the effect of ephedrine on the expression of interleukin-8（ IL-8） in human bronchial epithelial cells（ 16 HBE cells） and to explore the mechanism of ephedra in treatment of asthma. Methods The 16 HBE cells were randomly divided into three groups： group A,group B and group C. Groups A and B were respectively added with TNF-α（ 20 ng / m L） and TNF-α（ 20 ng / m L） ＋ ephedrine（ 300μg / m L） in vitro for culture,but no treatment in group C. After cells were incubated for 18 hours,IL-8 mRNA level was measured by real-time PCR and protein expression by Western blotting. The IL-8 in cell culture supernatant was quantified by ELISA. Results The mRNA and protein expression levels of IL-8 in 16 HBE cells of groups A and B were higher than those of group C（ B ﹥ A ﹥ C）,and the statistical difference was significant among the three groups（ all P 0. 05）. The protein expression of IL-8 in the supernatant of groups A and B was（ 27. 24 ± 1. 91） and（ 23. 62 ± 3. 40） pg / m L,which was higher than that of group C（ 17. 45 ± 2. 32）pg / m L（ all P 0. 05）. Significant difference was found in the IL-8 protein of the supernatant between groups A and B（ P 0. 05）. Conclusion Ephedrine can inhibit the expression and secretion of IL-8 in 16 HBE cells stimulated by TNF-α,which may be one of the mechanisms of ephedra in treatment of asthma.