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目的:探讨一氧化氮(NO)对香烟烟雾提取物(CSE)诱导的大鼠肺泡巨噬细胞(AM)中核因子κB(NF—κB)活化的影响及机 制。方法:将大鼠AM与不同浓度的NO前体左旋精氨酸(L-Arg)或iNOS特异性抑制剂N6-(1-亚氨乙基)赖氨酸(L-NIL)及CSE共同培养,用免疫细胞化学染色法检测NF-κB,用Western blot检测I-κB蛋白含量,用Griess法测定培养上清液中NO的水平。结果:CSE可使NF-κB活化细胞的百分率增加,I-κB的水平下降。加入CSE和低浓度L-Arg培养的AM,NF-κB活化细胞的百分率显著高于只加入CSE的AM;而I-κB的水平显著低于只加入CSE的AM。加入CSE和高浓度L-Arg培养的AM,NF-κB活化细胞的百分率显著低于只加入CSE的AM,而I-κB的水平无显著变化。加入CSE和不同浓度的L-NIL培养的AM,NF-κB活化细胞的百分率显著低于只加入CSE的AM;而I-κB的水平则显著高于只加入CSE的AM,并呈浓度依赖(P<0.01)。结论:内源性NO对香烟烟雾所致NF-κB的活化具有双向调控作用。
Objective: To investigate the effect of nitric oxide (NO) on activation of nuclear factor κB (NF-κB) in rat alveolar macrophages (AM) induced by cigarette smoke extract (CSE) and its mechanism. METHODS: Rat AM was co-cultured with different concentrations of NO precursor L-Arg or iNOS specific inhibitor N6- (1-iminoethyl) lysine (L-NIL) and CSE NF-κB was detected by immunocytochemical staining, the content of I-κB protein was detected by Western blot, and the level of NO in culture supernatant was determined by Griess method. Results: CSE increased the percentage of activated NF-κB cells and decreased the level of I-κB. The percentage of activated NF-κB-activated cells in AM with CSE and low concentration of L-Arg was significantly higher than that with CSE only, while the level of I-κB was significantly lower than AM with only CSE added. The percentage of NF-κB-activated cells in AMs incubated with CSE and high concentration of L-Arg was significantly lower than that of AMs with CSE only, while there was no significant change in the level of I-κB. The percentage of activated NF-κB-activated cells in CS cultured with L-NIL and different concentration of L-NIL was significantly lower than that of AM with only CSE added, while the level of I-κB was significantly higher than that of AM with CSE alone P <0.01). CONCLUSION: Endogenous nitric oxide can bidirectionally regulate the activation of NF-κB induced by cigarette smoke.