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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。
Objective: To observe the effect of low dose of theophylline on the expression of NF-κB and ICAM-1 in cultured rat airway smooth muscle cells and to explore the effect of low dose of theophylline on airway anti-inflammation Possible mechanism of action. Methods: Airway smooth muscle cells of normal rats were cultured in vitro. The cultured cells were randomly divided into three groups: blank control group, tumor necrosis factor-α (TNF-α) -stimulated group and theophylline intervention group. The expression of NF-κB and ICAM-1 in the airway smooth muscle cells of rats stimulated with TNF-α were detected by immunohistochemical SP method after adding different concentrations of theophylline for 4 h, and incubated with the same concentration of theophylline for different time. 1 expression changes. Results: The difference between NF-κB activity and ICAM-1 expression in TNF-α stimulation group and blank control group was significant (P <0.05). The NF-κB activity and ICAM-1 expression in airway smooth muscle cells stimulated by TNF-α were decreased after adding 5, 10 and 15 mg / L theophylline for 4 h. Compared with the TNF-α stimulation group, there was no significant difference in the 5 mg / L group (P> 0.05). The differences between the two groups were significant (P <0.05). Theophylline intervention group within each comparison, the difference was significant (P <0.05). The airway smooth muscle cells of TNF-α-stimulated rats were treated with 10 mg / L theophylline for 2,4,24 h. Compared with the TNF-α stimulation group, theophylline treatment group differences were significant (P <0.05). However, there was no significant difference between different treatment time in theophylline treatment group (P> 0.05). Conclusion: Theophylline may reduce the infiltration of inflammatory cells in the airway by inhibiting NF-κB activity and ICAM-1 expression in airway smooth muscle cells.