NALP3及NF-κB信号通路在氧化应激反应中的作用及阿魏酸钠的干预效应

Sodium Ferulate Attenuates Oxidative Stress Induced Inflammation via Suppressing NALP3 and NF-κB Signal Pathway

中文关键词:  H2O2 人肺上皮细胞A549 NALP3 NF-κB IL-1β 阿魏酸钠

英文关键词:H2O2 Human lung epithelial cells A549 NACHT-PYD-containing protein 3 Nuclear factor kappa B Interleukin-1beta Sodium ferulate

中文摘要:

      目的 检测氧化应激时人肺上皮细胞（A549）炎性复合体NALP3、核转录因子-kappa B （NF-κB）基因和蛋白的表达，探讨阿魏酸钠对其的干预作用及可能的作用机制。方法 培养A549细胞，分为对照组、 H2O2（100 μmol/L）应激组、NF-κB阻断剂组（PDTC + H2O2组）、阿魏酸钠干预组（阿魏酸钠+H2O2组），其中PDTC(100 μmol/L)和阿魏酸钠（400 μg/mL）预处理30 min后加入H2O2（100 μmol/L），培养2 h后，采用实时荧光定量-PCR （qRT-PCR）检测NALP3、NF-κB（P65） mRNA的表达； Western blot检测NALP3、IκBα蛋白的表达；酶联免疫吸附（ELISA）测定细胞上清中白介素-1β（IL-1β）的表达。结果 与对照组比较，H2O2 可使A549细胞NALP3 mRNA、NALP3蛋白水平表达增强，NF-κB（P65） mRNA表达上调、IκBα降解增强，IL-1β分泌增加（P均<0.05）；而阿魏酸钠及NF-κB阻断剂PDTC可抵抗H2O2对A549细胞的作用，与H2O2应激组比较，下调NALP3 mRNA、NALP3蛋白、NALP3、NF-κB（P65） mRNA表达，IκBα降解减少，IL-1β分泌下降（P均<0.05），并且阿魏酸钠与NF-κB阻断剂PDTC上述作用差异无统计学意义。结论 阿魏酸钠可能通过抑制NF-κB的活化，减少NALP3及IL-1β的表达，从而减轻氧化应激导致的炎症反应。

英文摘要:

      Objective To study the effects of sodium ferulate on inflammation in human lung epithelial cells (A549) under oxidative stress and itsinfluence onthe expression of inflammasome NACHT-PYD-containing protein 3 (NALP3) and nuclear factor kappa B (NF-κB). Methods Human lung epithelial cells A549 cultured In vitro were divided into 4 groups, including control group, H ₂ O ₂ (100 μmol/L) stress group, NF-κB blockers group (PDTC 100 μmol/L+ H ₂ O ₂ 100 μmol/L), sodium ferulate （SF） intervention group (SF 400 μg/mL+H ₂ O ₂ 100 μmol/L). The expression of NALP3, IκBα protein were evaluated by Western blot, while mRNA levels of NALP3,NF-κB（P65） were measured by qRT-PCR.The level of interleukin-1beta (IL-1β) were detected by ELISA. Results H ₂ O ₂ not only increased the mRNA and protein expression levels of NALP3, but also enhanced the secretion of IL-1β in human lung epithelial cells A549 (P<0.05) when compared with control group.NF-κB blockers PDTC and sodium ferulateresisted the effects of H ₂ O ₂ on A549 cells, that decreased the mRNA and protein expression of NALP3 and the mRNA expression of NF-κB (P65), reduced the degeneration of IκBα and the secretion of IL-1β (P<0.05) when compared to H ₂ 2O ₂ stress group. Conclusion SF may reduce the expression of NALP3 and IL-1β by inhibiting NF-κB, so as to reduce the inflammation caused by oxidative stress.