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目的研究左旋精氨酸对NCI-H292细胞中白细胞介素6(IL-6)和IL-8的影响。方法细胞共分为5组:最低需要培养基(MEM)组、MEM+A组(MEM+左旋精氨酸)、RPMI组(RPMI1640培养基,含1 mmol.L-1精氨酸)、常规培养基组(精氨酸含量>1 mmol.L-1)、地塞米松组(MEM+10μg.L-1地塞米松)。通过TNF-α和脂多糖刺激NCI-H292细胞建立哮喘气道炎症微环境,观察不同浓度TNF-α和脂多糖对IL-6和IL-8释放的影响,并比较各组间IL-6和IL-8水平。采用ELISA法测定细胞IL-6和IL-8释放水平,斑点印迹法测定其mRNA水平。结果 IL-6和IL-8水平与TNF-α的浓度正相关(r=0.644、0.715,均P<0.01)。不论有无脂多糖或TNF-α的刺激,MEM组IL-6和IL-8水平高于MEM+A组、RPMI组和地塞米松组(P<0.01)。MEM组IL-6水平高于常规培养基组(P<0.01),TNF-α刺激下IL-8水平高于常规培养基组(P<0.01),脂多糖刺激下2组IL-8水平相当(P>0.05)。地塞米松组IL-6和IL-8水平与MEM+A组相当(P>0.05)。细胞培养4、8、24 h,MEM+A组IL-6 mRNA和IL-8 mRNA表达显著低于MEM组(P<0.05)。结论一定浓度左旋精氨酸可抑制NCI-H292细胞IL-6和IL-8的释放,可能通过降低IL-6和IL-8转录活性引起,在哮喘治疗中具有潜在的应用价值。
Objective To investigate the effects of L-arginine on interleukin-6 (IL-6) and IL-8 in NCI-H292 cells. Methods The cells were divided into five groups: MEM group, MEM + A group, RPMI group (RPMI1640 medium containing 1 mmol.L-1 arginine), routine culture Base group (arginine content> 1 mmol.L-1), dexamethasone group (MEM + 10 μg.L-1 dexamethasone). The airway inflammation microenvironment of NCI-H292 cells was stimulated by TNF-α and lipopolysaccharide, and the effects of different concentrations of TNF-α and lipopolysaccharide on the release of IL-6 and IL-8 were observed. The levels of IL-6 and IL- IL-8 levels. The levels of IL-6 and IL-8 in cells were measured by ELISA, and the mRNA levels were determined by dot blotting. Results The levels of IL-6 and IL-8 positively correlated with the concentration of TNF-α (r = 0.644, 0.715, all P <0.01). The levels of IL-6 and IL-8 in MEM group were higher than those in MEM + A group, RPMI group and dexamethasone group (P <0.01), with or without stimulation of lipopolysaccharide or TNF-α. The level of IL-6 in MEM group was higher than that in routine medium group (P <0.01). The level of IL-8 in TNF-α group was higher than that in routine medium group (P <0.01) (P> 0.05). The levels of IL-6 and IL-8 in dexamethasone group were similar to those in MEM + A group (P> 0.05). At 4, 8 and 24 h after culture, the expression of IL-6 mRNA and IL-8 mRNA in MEM + A group was significantly lower than that in MEM group (P <0.05). Conclusion L-arginine can inhibit the release of IL-6 and IL-8 in NCI-H292 cells, which may be caused by decreasing the transcriptional activity of IL-6 and IL-8 and has potential value in the treatment of asthma.