氨茶碱对TNF-α诱导下大鼠气道平滑肌细胞中RANTES的影响

Effects of Theophylline on the Expression of RANTES in the Airway Smooth Muscle Cells of Rats Induced by TNF-alpha

目的:观察临床治疗范围内不同浓度的氨茶碱对肿瘤坏死因子-α(TNF-α)诱导下大鼠气道平滑肌细胞(ASMC)中活化正常T细胞表达和分泌的调节物(RANTES)的影响及相应浓度的氨茶碱作用下细胞内环磷酸腺苷(cAMP)的变化,了解趋化因子在哮喘发病机制中的作用,进一步阐明茶碱类药物在治疗哮喘中的抗炎机制。方法:将分离的大鼠ASMC进行传代培养,培养液中加入TNF-α(终浓度为10μg/L)诱导后,再加入不同浓度的氨茶碱,用ELISA方法检测上清液中RANTES的浓度,用放射免疫法检测培养液中cAMP的浓度。结果:经TNF-α诱导后,不加氨茶碱组中大鼠ASMC上清液中RANTES的浓度最高,为(13.799±0.250)×10-9g/L,不同浓度氨茶碱作用后,RANTES的浓度均有所减少,其中5(即氨茶碱终浓度为5 mg/L,下同),7.5,10,12.5,15和17.5 mg/L组RANTES的浓度分别为(13.088±0.252),(12.555±0.100),(10.425±0.502),(7.943±0.500),(5.465±0.500),(4.581±0.25)ng/L。除5 mg/L组和对照组相比无显著性差异(P>0.05)外,其余氨茶碱组与对照组组差异均具有统计学意义(P<0.05)。各氨茶碱组之间相比较,除5 mg/L组和7.5 mg/L组相比无显著性意义(P>0.05)外,其余氨茶碱组两两相比均有统计学意义(P<0.05)。TNF-α诱导下,细胞内cAMP的浓度随着氨茶碱浓度的增加而增高。结论:茶碱类药物可抑制大鼠ASMC中RANTES的产生,并可能呈浓度依赖性,而且这种抑制作用与细胞内cAMP的变化有一定的关系。

Objective： To explore the roles of theophylline on the expression of RANTES and cAMP in the airway smooth muscle cells （ASMCs） of rats induced by TNF-α, and to investigate RANTES and theophylline in the pathogenesis of bronchial asthma. Methods： The trachea smooth muscle was obtained from Wistar rat, and ASMCs were dissected, purified, and cultured in medium. Then were treated with TNF-α （the final concentration was 10 μg/L） and different concentration of theophylline and ELISA method was used to determine the concentration of RANTES. The level of cAMP was measured by radioimmunoassay. Results： After treated with both TNF-α and theophylline, the expression of RANTES in rat ASMCs was lower than in the group without theophylline, and the higher concentration of theophylline, the lower of expression of RANTES.Except the 5 mg/L group, a significant positive correlation existed between the theophylline groups and control group（P〈0.05）. Comparing among different dose of theophylline group, the 5 mg/L group had no significant difference with 7.5 mg/L（P〉0.05）. Conclusion： Theophylline, of which the concentration is above 7.5 mg/L, can inhibit the expression of RANTES in ASMCs of rats. And the inhibition of theophylline on RANTES was related with cAMP.