小剂量茶碱对大鼠气道平滑肌细胞表达NF-κB与ICAM-1的影响

目的:观察小剂量茶碱对培养的大鼠气道平滑肌细胞表达核因子κB(NF-κB)与细胞间黏附分子1(ICAM-1)的影响,探讨小剂量茶碱在气道发挥抗炎作用的可能机制。方法:体外培养正常大鼠鼠婴的气道平滑肌细胞,将培养出的细胞随机分为3组:空白对照组、肿瘤坏死因子α(TNF-α)刺激组、茶碱干预组。采用免疫组织化学SP法检测经TNF-α刺激的大鼠的气道平滑肌细胞中加入不同浓度的茶碱共同培养4 h,以及加入同一浓度茶碱共同培养不同时间,NF-κB活性和ICAM-1表达的变化。结果:TNF-α刺激组与空白对照组的NF-κB活性和ICAM-1表达相比较,差异均有显著性(P<0.05)。经TNF-α刺激的大鼠的气道平滑肌细胞中加入5,10,15 mg/L的茶碱共同培养4 h,NF-κB活性和ICAM-1表达均有所下降。与TNF-α刺激组相比,5 mg/L组无统计学差异(P>0.05),余两组差异均有显著性(P<0.05)。且茶碱干预组内两两比较,差异均有显著性(P<0.05)。经TNF-α刺激的大鼠的气道平滑肌细胞中加入10 mg/L茶碱共同培养2,4,24 h。与TNF-α刺激组相比,茶碱处理组的差异均有显著性(P<0.05)。但茶碱处理组内不同处理时间两两比较,差异无显著性(P>0.05)。结论:茶碱可能通过抑制气道平滑肌细胞NF-κB活性和ICAM-1的表达,进而减少炎症细胞在气道的浸润。

Effect of Low-dose Theophylline on Expression of NF-kappa B and ICAM-1 in Airway Smooth Muscle Cells of Rats

Objective: To observe the effect of low-dose theophylline on expression of NF-κB and ICAM-1 in cutured airway smooth muscle cells of rats,and to understand the mechanisms of low-dose theophylline on anti-inflammation in airway.Methods: Airway smooth muscles of baby rats were cultured in vitro.The cutured cells were divided into three groups randomly:the control group,the TNF-α stimulation group and the theophylline treatment group.Airway smooth muscle cells of the theophylline treatment group were stimulated by TNF-α in advance,then they were added in 5 mg/L,10 mg/L and 15 mg/L theophylline respectively,cutured for 4 hours or added in 10 mg/L theophylline,cutured for 2,4 and 24 hours respectively.Immunohistochemical stain were used to evaluate the activity of NF-κB and the expression of ICAM-1.Results: The level of nuclear NF-κB protain expression and ICAM-1 protain expression all showed a significant difference between the TNF-α stimulation group and the control group(P<0.05).The cells stimulated by TNF-α were cultured with 5 mg/L,10 mg/L and 15 mg/L theophylline for 4 hours.Compared with the TNF-α stimulation group,these three groups all showed a significant difference(all P<(0.05)) except for 5 mg/L group.The cells stimulated by TNF-α were cultured with 10 mg/L theophylline for 2,4 and 24 hours,no significant difference(all P>0.05) was observed among them.Compared with the TNF-α stimulation group,these three groups all showed a significant difference(all P<0.05).(Conclusion:) Theophylline can reduce the infiltration of inflammatory cells through inhibiting the activity of NF-κB and the expression of ICAM-1 in airway smooth muscle cells.