氨茶碱对哮喘缓解期儿童TH相关细胞因子调节作用的实验研究

Modulatory effect of aminophylline on the cytokine excreted by TH of asthmatic children during remission period.

目的:探讨氨茶碱对哮喘缓解期患儿抗炎作用免疫机理。方法:采集20例缓解期哮喘儿童PBMCs与氨茶碱共同培养,按不同浓度(0、5、10、20mg/L)分为哮喘对照组(ACG)、处理组(ATG1)、ATG2和ATG3,同时设立正常对照组(NCG)。采用ELISA检测培养24h后各组PBMCs上清液中IL-4、IL-10、IFN-γ浓度和血清IgE浓度,同时计数外周血EOS。结果:哮喘缓解期患儿EOS和IgE水平均明显高于NCG;ACG组IL-4、IL-10明显高于NCG,IFN-γ明显低于NCG;20mg/L氨茶碱能显著抑制IL-4、IL-10分泌,提高IFN-γ的分泌,逆转IL-4/IFN-γ比值。结论:在哮喘缓解期患儿仍然存在TH2亚群功能亢进;20mg/L的氨茶碱可以通过纠正TH1和TH2细胞因子的失衡状态而发挥抗炎作用。

Objective： To explore the mechanism of anti-inflammatory action of aminophylline in asthmatic children. Methods： PBMCs, separated from vein blood of 20 asthmatic children, wero cultivated together with aminophylline and then divided into four groups： ACG, ATG1, ATG2, ATG3, according to aminophylline concentrations（0, 5, 10 and 20 mg/L）. At the same time, a control group of NCG was set up. The serum concentration of IgE, the concentrations of IL-4, IL-10 and IFN-γ/from supernatant of PBMCs were cultivated 24 h after the test of ELISA, meanwhile the number of EOS in vein blood was counted. Results： EOS and IgE in asthma groups were significantly higher than those of NCG; IL-4 level in ACG was significandy higher than in NCG, while IFN-γ level was significantly lower than that of NCG; IL-4/IFN-γ ratio was significantly higher than that of NCG; from ACG to ATG3, IL-4 concentration was gradually decreasing. Aminophylline mediated a slight inhibition which was only obvious at the concentrion of 20 mg/L （P 〈 0.01）; the IFN-γ level of ATG3 was significantly higher than that of ACG; anfinophylline reduced IL-4/IFN-γratio, but only significantly in aminophylline concentration of 20 mg/L. There was significantly positive correlation between EOS and IL-4/IFN-γ ratio and negative correlation between IL-4 and IFN-γ in ACG. Conclusions： There was TH2 subset hyperfunction during asthma remission period, characterized by higher IL-4 level, IL-10 level, IL-4/IFN-γ ratio. These data suggest that cytokines expressions of TH2 be inhibited at therapeutic concentration superior limit （20 rag/ L） of aminophylline, and its suppression to TH2 cytokines is likely to be an important mechanism of anti-inflammation at the therapeutic concentration of aminophylline in asthma.