欢迎来到《四川大学学报(医学版)》

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

  • 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。

     

    Abstract: 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.

     

/

返回文章
返回