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

作　　者：何高燕 谢敏[1] 高艳[1] 黄建戈[1] 

机构地区：[1]四川大学华西医院呼吸内科,成都610041

出　　处：《四川大学学报（医学版）》2015年第3期367-371,共5页Journal of Sichuan University(Medical Sciences)

基　　金：四川省科技厅科技支撑计划项目(No.2013SZ0001)资助

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(100μmol/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(100μmol/L)和阿魏酸钠(400μg/mL)预处理30min后加入H2O2(100μmol/L),培养2h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NALP3mRNA、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NALP3mRNA、NALP3蛋白、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。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 4groups,including control group,H2O2（100μmol/L）stress group,NF-κB blockers group（PDTC100μmol/L＋ H2O2100μmol/L）,sodium ferulate（SF）intervention group（SF 400μg/mL＋H2O2100μmol/L）.The expression of NALP3,IκBαprotein 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 H2O2 not only increased the mRNA and protein expression levels of NALP3,but also enhanced the secretion of IL-1βin human lung epithelial cells A549（P〈0.05）when compared with control group.NF-κB blockers PDTC and sodium ferulateresisted the effects of H2O2 on A549cells,that decreased the mRNA and protein expression of NALP3 and the mRNA expression of NF-κB（P65）,reduced the degeneration of IκBαand the secretion of IL-1β（P〈0.05）when compared to H2O2 stress group.Conclusion SF may reduce the expression of NALP3 and IL-1βby inhibiting NF-κB,so as to reduce the inflammation caused by oxidative stress.

关 键 词：H2O2 人肺上皮细胞A549 NALP3 NF-κB IL-1β 阿魏酸钠 

分 类 号：R96[医药卫生—药理学]