DDX3X promotes endoplasmic reticulum protein reprogramming via interaction with ERN1 to amplify cerebral ischemia/reperfusion injury

Abstract Background Reperfusion therapy is a critical intervention to salvage acute cerebral ischemia. However, reperfusion itself induces cerebral ischemia/reperfusion (I/R) injury. Recent evidence suggests that DEAD-box helicase 3, X-linked (DDX3X) and Endoplasmic Reticulum to Nucleus Signaling 1...

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Main Authors: QiaoHui Du, YuQian Liu, JingLong Guo
Format: Article
Language:English
Published: BMC 2025-07-01
Series:European Journal of Medical Research
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Online Access:https://doi.org/10.1186/s40001-025-02813-y
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author QiaoHui Du
YuQian Liu
JingLong Guo
author_facet QiaoHui Du
YuQian Liu
JingLong Guo
author_sort QiaoHui Du
collection DOAJ
description Abstract Background Reperfusion therapy is a critical intervention to salvage acute cerebral ischemia. However, reperfusion itself induces cerebral ischemia/reperfusion (I/R) injury. Recent evidence suggests that DEAD-box helicase 3, X-linked (DDX3X) and Endoplasmic Reticulum to Nucleus Signaling 1 (ERN1) play pivotal roles in various diseases, particularly those affecting the nervous system. However, the exact roles of DDX3X and ERN1 in cerebral I/R injury remain unclear. Methods We used Sprague–Dawley (SD) rats subjected to middle cerebral artery occlusion (MCAO) surgery and PC12 cells exposed to oxygen–glucose deprivation/reoxygenation (OGD/R) to establish cerebral I/R injury models. Western blotting, immunohistochemistry, and co-immunoprecipitation were employed to assess the expression and interaction of DDX3X and ERN1. CCK-8 assay and flow cytometry were used to evaluate cell viability and apoptosis. Western blotting was performed to measure key endoplasmic reticulum stress (ERS) markers and downstream targets. Pro-inflammatory cytokines were quantified by ELISA. Results DDX3X and ERN1 were significantly upregulated in both OGD/R-treated PC12 cells and MCAO rat brain tissue. Importantly, DDX3X was shown to positively regulate ERN1 expression, leading to increased apoptosis, ERS, oxidative stress, and inflammation in both cellular and animal models. These processes collectively exacerbated cerebral injury, reinforcing the pathological role of DDX3X and ERN1 interaction in the progression of cerebral I/R injury. Conclusion DDX3X enhances cerebral I/R injury by interacting with ERN1, which triggers the reprogramming of endoplasmic reticulum protein responses, amplifying ERS, apoptosis, and inflammation.
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spelling doaj-art-2b2d962ace254400a4552c3a2527c8ea2025-08-20T03:45:24ZengBMCEuropean Journal of Medical Research2047-783X2025-07-0130111310.1186/s40001-025-02813-yDDX3X promotes endoplasmic reticulum protein reprogramming via interaction with ERN1 to amplify cerebral ischemia/reperfusion injuryQiaoHui Du0YuQian Liu1JingLong Guo2Health Management Center, Shandong Provincial Qianfoshan Hospital, The First Affiliated Hospital of Shandong First Medical UniversityHealth Management Center, Shandong Provincial Qianfoshan Hospital, The First Affiliated Hospital of Shandong First Medical UniversityDepartment of Hepatobiliary Surgery, Shandong Provincial Third Hospital, Shandong UniversityAbstract Background Reperfusion therapy is a critical intervention to salvage acute cerebral ischemia. However, reperfusion itself induces cerebral ischemia/reperfusion (I/R) injury. Recent evidence suggests that DEAD-box helicase 3, X-linked (DDX3X) and Endoplasmic Reticulum to Nucleus Signaling 1 (ERN1) play pivotal roles in various diseases, particularly those affecting the nervous system. However, the exact roles of DDX3X and ERN1 in cerebral I/R injury remain unclear. Methods We used Sprague–Dawley (SD) rats subjected to middle cerebral artery occlusion (MCAO) surgery and PC12 cells exposed to oxygen–glucose deprivation/reoxygenation (OGD/R) to establish cerebral I/R injury models. Western blotting, immunohistochemistry, and co-immunoprecipitation were employed to assess the expression and interaction of DDX3X and ERN1. CCK-8 assay and flow cytometry were used to evaluate cell viability and apoptosis. Western blotting was performed to measure key endoplasmic reticulum stress (ERS) markers and downstream targets. Pro-inflammatory cytokines were quantified by ELISA. Results DDX3X and ERN1 were significantly upregulated in both OGD/R-treated PC12 cells and MCAO rat brain tissue. Importantly, DDX3X was shown to positively regulate ERN1 expression, leading to increased apoptosis, ERS, oxidative stress, and inflammation in both cellular and animal models. These processes collectively exacerbated cerebral injury, reinforcing the pathological role of DDX3X and ERN1 interaction in the progression of cerebral I/R injury. Conclusion DDX3X enhances cerebral I/R injury by interacting with ERN1, which triggers the reprogramming of endoplasmic reticulum protein responses, amplifying ERS, apoptosis, and inflammation.https://doi.org/10.1186/s40001-025-02813-yProtein interactionEndoplasmic reticulum reprogrammingCerebral ischemia/reperfusion injuryGene silencing
spellingShingle QiaoHui Du
YuQian Liu
JingLong Guo
DDX3X promotes endoplasmic reticulum protein reprogramming via interaction with ERN1 to amplify cerebral ischemia/reperfusion injury
European Journal of Medical Research
Protein interaction
Endoplasmic reticulum reprogramming
Cerebral ischemia/reperfusion injury
Gene silencing
title DDX3X promotes endoplasmic reticulum protein reprogramming via interaction with ERN1 to amplify cerebral ischemia/reperfusion injury
title_full DDX3X promotes endoplasmic reticulum protein reprogramming via interaction with ERN1 to amplify cerebral ischemia/reperfusion injury
title_fullStr DDX3X promotes endoplasmic reticulum protein reprogramming via interaction with ERN1 to amplify cerebral ischemia/reperfusion injury
title_full_unstemmed DDX3X promotes endoplasmic reticulum protein reprogramming via interaction with ERN1 to amplify cerebral ischemia/reperfusion injury
title_short DDX3X promotes endoplasmic reticulum protein reprogramming via interaction with ERN1 to amplify cerebral ischemia/reperfusion injury
title_sort ddx3x promotes endoplasmic reticulum protein reprogramming via interaction with ern1 to amplify cerebral ischemia reperfusion injury
topic Protein interaction
Endoplasmic reticulum reprogramming
Cerebral ischemia/reperfusion injury
Gene silencing
url https://doi.org/10.1186/s40001-025-02813-y
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AT jinglongguo ddx3xpromotesendoplasmicreticulumproteinreprogrammingviainteractionwithern1toamplifycerebralischemiareperfusioninjury