DDX3X Deficiency Attenuates Pyroptosis Induced by Oxygen-glucose Deprivation/Reoxygenation in N2a Cells

作者全名："Liu, Yong; Gui, Yanlin; Tang, Hao; Yu, Jianping; Yuan, Zhengzhou; Liu, Lei; Ma, Xuntai; Li, Changqing"

作者地址："[Liu, Yong; Li, Changqing] Chongqing Med Univ, Dept Neurol, Affiliated Hosp 2, Chongqing 400010, Peoples R China; [Liu, Yong; Yu, Jianping; Yuan, Zhengzhou; Liu, Lei; Ma, Xuntai] Chengdu Med Coll, Dept Neurol, Affiliated Hosp 1, Chengdu 610500, Sichuan, Peoples R China; [Gui, Yanlin] Chengdu Med Coll, Chengdu 610500, Sichuan, Peoples R China; [Tang, Hao] Chongqing Med Univ, Dept Neurol, Affiliated Hosp 1, Chongqing 400016, Peoples R China"

通信作者："Li, CQ (通讯作者)，Chongqing Med Univ, Dept Neurol, Affiliated Hosp 2, Chongqing 400010, Peoples R China."

来源：CURRENT NEUROVASCULAR RESEARCH

ESI学科分类：NEUROSCIENCE & BEHAVIOR

WOS号：WOS:001078214200005

JCR分区：Q3

影响因子：2

年份：2023

卷号：20

期号：2

开始页：197

结束页：206

文献类型：Article

关键词：DDX3X; NLRP3; OGD/R; pyroptosis; cell viability; immunofluorescence

摘要："Background: NOD-like receptor protein 3 (NLRP3) inflammasome-mediated pyroptosis is strongly related to cerebral ischemia/reperfusion (I/R) injury. DDX3X, the DEAD-box family's ATPase/RNA helicase, promotes NLRP3 inflammasome activation. However, whether DDX3X deficiency attenuates NLRP3 inflammasome-mediated pyroptosis induced by cerebral I/R injury.Objectives: This study investigated whether DDX3X deficiency attenuates NLRP3 inflammasome-mediated pyroptosis in N2a cells after oxygen-glucose deprivation/ reoxygenation (OGD/R) treatment.Methods: In vitro model of cerebral I/R injury, mouse neuro2a (N2a) cells subjected to OGD/R were treated with the knockdown of DDX3X. Cell counting kit-8 (CCK-8) assay and Lactate dehydrogenase (LDH) cytotoxicity assay were conducted to measure cell viability and membrane permeability. Double immunofluorescence was performed to determine the pyroptotic cells. Transmission electron microscopy (TEM) was used to observe morphological changes of pyroptosis. Pyroptosis-associated proteins were analyzed by Western blotting.Results: The OGD/R treatment reduced cell viability, increased pyroptotic cells and released LDH compared to the control group. TEM showed membrane pore formation of pyroptosis. Immunofluorescence showed that GSDMD was translocated from the cytoplasm to the membrane after OGD/R treatment. Western blotting showed that the expression of DDX3X, and pyroptosis-related proteins (NLRP3, cleaved-Caspase1, and GSDMD-N) were increased after OGD/R treatment. Nevertheless, DDX3X knockdown markedly improved cell viability and reduced LDH release, expression of pyroptosis-related proteins, and N2a cells pyroptosis. DDX3X knockdown significantly inhibited membrane pore formation and GSDMD translocation from cytoplasm to membrane.Conclusion: This research demonstrates for the first time that DDX3X knockdown attenuates OGD/R-induced NLRP3 inflammasome activation and pyroptosis, which implies that DDX3X may become a potential therapeutic target for cerebral I/R injury."

基金机构：Declared none.

基金资助正文：Declared none.