麻黄碱对TNF-α诱导人支气管上皮细胞eotaxin表达的影响

Effect of ephedrine on expression of eotaxin in human bronchial epithelial cells stimulated by tumor necrosis factor-α

目的观察麻黄碱对肿瘤坏死因子-α(TNF-α)诱导的人支气管上皮细胞(16HBE)中嗜酸性粒细胞趋化因子(eotaxin)表达的影响,探讨中药麻黄治疗哮喘的机制。方法将体外培养的16HBE细胞随机分为对照组、TNF-α刺激组(TNF-α20ng/mL)、TNF-α+麻黄碱组(TNF-α20ng/mL+麻黄碱300μg/mL),每组细胞均设3个复孔培养18h;荧光定量PCR检测各组细胞eotaxin mRNA的表达;免疫细胞化学染色法和Western blot检测各组细胞eotaxin蛋白的表达;双抗体夹心酶联免疫吸附法(ELISA)测定各组细胞培养上清液中eotaxin的浓度。结果与对照组比较,TNF-α刺激组上皮细胞eotaxin mRNA及蛋白表达、细胞培养上清液中eotaxin的浓度明显增高,差异均有统计学意义(P<0.01);与TNF-α刺激组比较,TNF-α+麻黄碱组以上各项指标均明显降低,差异有统计学意义(P<0.01)。结论麻黄碱能抑制前炎症因子TNF-α诱导的16HBE中eotaxin的表达及分泌,这可能是中药麻黄治疗哮喘的机制之一。

Objective To observe the effect of ephedrine on the expression of eotaxin in human bronchial epithelial cells（16HBE）stimulated by tumor necrosis factor-α（TNF-α）and to explore the mechanism of Chinese medicine ephedra in treating asthma.Methods The in vitro cultured 16 HBE were randomly divided into the control group,TNF-αstimulation group（TNF-α20ng/mL）and TNF-αplus ephedrine group（TNF-α20ng/mL plus ephedrine 300μg/mL）.Three complex holes in each group were set to culture for 18 h,the eotaxin mRNA expression was measured by real time fluorescent quantified PCR and protein level was detected by immunocytochemical stain and Western blot.The eotaxin concentration in cells culture supernatant was quantified by ELISA.Results Compared with the the control group,the expression level of eotaxin mRNA and protein,and the concentration of eotaxin in cell culture supernatant in the TNF-αstimulation group were increased obviously,there being statisticaly significant difference between them（P〈0.01）;however,all above these parameters in the TNF-αplus ephedrine group showed decreased obviously as compared with the TNF-αgroup,the difference between them was statistically significant（P〈0.01）.Conclusion Ephedrine can inhibit the expression and secretion of eotaxin in TNF-αinduced 16 HBE inflammatory model,which may be one of the mechanisms of Chinese medicine ephedra in treating asthma.