通过NF-κB调控蛋白激酶SPAK通路促进NR8383细胞炎症介质释放及氧化应激发生  

NF-κB regulates the release of inflammatory mediators and occurrence of oxidative stress in NR8383 cells through SPAK pathway

作　　者：陈妍 薄丽艳 李聪聪 Chen Yan;Bo Liyan;Li Congcong(Department of Respiratory and Critical Care Medicine, General Hospital of Northern Theater Command, Shenyang 110016, China;Department of Respiratory and Critical Care Medicine, Xi′an chest Hospital of Shanxi Province 710100, China)

机构地区：[1]中国人民解放军北部战区总医院呼吸与危重症医学科,沈阳110001 [2]陕西省西安市胸科医院呼吸与危重症医学科,西安710100

出　　处：《中华肺部疾病杂志（电子版）》2022年第2期146-150,共5页Chinese Journal of Lung Diseases(Electronic Edition)

基　　金：国家自然科学基金青年项目(81900083&81800076);中国博士后科学基金第65批面上资助(2019M653911)。

摘　　要：目的分析SPAK在高渗透压应激所致大鼠肺泡世噬细胞系(NR8383)细胞炎症介质释放及氧化应激发生中的作用。方法在细胞培养液中加入核因子κB(nuclear factorκappa-B,NF-κB)的抑制剂吡咯烷二硫代氨基甲盐酸(1-Pyrrolidinecarbodithioic acid,PDTC)阻断其磷酸化,检测其对高渗刺激引起的SPAK表达升高的抑制作用。通过siRNA干扰技术下调SPAK的表达,检测NR8383细胞炎症介质TNF-α和IL-1的分泌以及MPO活性和LDH释放,阐明SPAK在细胞应对高渗透压刺激时的调控作用。结果高渗透压刺激可以导致NR8383细胞中的NF-κB的磷酸化并上调SPAK蛋白表达,而通过对NR8383细胞进行PDTC预处理,NF-κB的磷酸化及SPAK蛋白表达水平可以显著受到抑制。而SPAK表达下调可以减轻高渗刺激所致的炎症因子释放增加,降低MPO活性以及H_(2)O_(2)浓度。结论磷酸化NF-κB可以调控SPAK转录及翻译,而SPAK可以通过促进炎性介质释放、诱导氧化应激参与到高渗透压刺激的调控中。Objective To investigate the role of STE20/SPS1-related proline/alanine-rich kinase(SPAK)in the release of inflammatory mediators and occurrence of oxidative stress in rat alveolar macrophage cells(NR8383)cells exposed hyperosmotic stress.Methods 1-Pyrrolidinecarbodithioic acid(PDTC),a specific inhibitor of NF-κB,was added to inhibit its phosphorylating,and the increase of SPAK expression induced by hypertonic stimulation was also detected.After down regulating the expression of SPAK in NR8383 cells by siRNA interference technology,the secretion of TNF-αand IL-1,MPO activity and H_(2)O_(2) concentration were detected,which further elucidated the role of SPAK in hyperosmotic stress regulation.Results PDTC pretreatment significantly inhibited the phosphorylation of NF-κB and the expression of SPAK protein in NR8383 cells.However,the down-regulation of SPAK expression could reduce the increased release of inflammatory factors,MPO activity and H_(2)O_(2) concentration caused by hypertonic stimulation.Conclusion The transcription of SPAK is regulated by the activation of NF-κB,and plays an important role in promoting the inflammatory response,the release of inflammatory mediators and occurrence of oxidative stress.

关 键 词：大鼠肺泡世噬细胞 核因子-ΚB 高渗透压刺激 蛋白激酶SPAK 氧化应激 

分 类 号：R563[医药卫生—呼吸系统]