2. Academic Validation
  3. THBS1 in macrophage-derived exosomes exacerbates cerebral ischemia-reperfusion injury by inducing ferroptosis in endothelial cells

THBS1 in macrophage-derived exosomes exacerbates cerebral ischemia-reperfusion injury by inducing ferroptosis in endothelial cells

  • J Neuroinflammation. 2025 Feb 24;22(1):48. doi: 10.1186/s12974-025-03382-x.
Chang Liu 1 2 3 4,  Haijing Sui 2 5,  Zhixi Li 2 3 4 5,  Zhenyu Sun 2 5,  Chenglong Li 6,  Guangmin Chen 7,  Zhaoxue Ma 2 5,  Hang Cao 2 5,  Hongjie Xi 8 9
Affiliations

Affiliations

  • 1 Department of Anesthesiology, Harbin Medical University Cancer Hospital, Harbin, 150081, People's Republic of China.
  • 2 The Key Laboratory of Anesthesiology and Intensive Care Research of Heilongjiang Province, Harbin, 150001, People's Republic of China.
  • 3 The Key Laboratory of Myocardial Ischemia Organization, Chinese Ministry of Education, Harbin, 150001, People's Republic of China.
  • 4 State Key Laboratory of Frigid Zone Cardiovascular Diseases, Harbin, 150001, People's Republic of China.
  • 5 Department of Anesthesiology, Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, People's Republic of China.
  • 6 Department of Anesthesiology, Fourth Affiliated Hospital of Harbin Medical University, 37 Yiyuan Road, Harbin, 150001, People's Republic of China.
  • 7 Department of Anesthesiology, First Affiliated Hospital of Harbin Medical University, 199 Dazhi Road, Harbin, 150001, People's Republic of China.
  • 8 The Key Laboratory of Anesthesiology and Intensive Care Research of Heilongjiang Province, Harbin, 150001, People's Republic of China. xihongjie@hrbmu.edu.cn.
  • 9 Department of Anesthesiology, Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, People's Republic of China. xihongjie@hrbmu.edu.cn.
PMID: 39994679 DOI: 10.1186/s12974-025-03382-x
Abstract

Macrophages play a critical role in the development of acute ischemic stroke (AIS). Cerebral ischemia-reperfusion injury (CIRI) is a pivotal pathological process that exacerbates AIS, with exosomes act as crucial mediators. However, the effects and mechanisms of action of macrophage-derived exosomes on CIRI remain unclear. This study demonstrated that macrophage-derived exosomes induce endothelial Ferroptosis and barrier disruption during CIRI. Through proteomic Sequencing and the reanalysis of transcriptomic and single-cell Sequencing data, thrombospondin-1 (THBS1) was identified as a key exosomal molecule. Elevated THBS1 was observed in exosomes and monocytes from the peripheral blood of patients with AIS in oxygen-glucose deprivation/reoxygenation (OGD/R)-stimulated THP-1 and RAW264.7, in their secreted exosomes, and in macrophages within the brains of transient middle cerebral artery occlusion (tMCAO) mice. Additionally, THBS1 expression in exosomes was positively correlated with vascular barrier injury biomarkers, including MMP-9 and S100B. Modulation of THBS1 in macrophage-derived exosomes affected exosome-induced Ferroptosis in endothelial cells. The mechanism by which THBS1 binds directly to OTUD5 and promotes GPX4 ubiquitination was elucidated using RNA interference, adeno-associated virus transfection, and endothelial-specific Gpx4 knockout mice. High-throughput screening of small-molecule compounds targeting THBS1 was performed. Molecular docking, molecular dynamics simulations, and cellular thermal shift assays further confirmed that salvianolic acid B (SAB) has a potent binding affinity for THBS1. SAB treatment inhibited the interaction between THBS1 and OTUD5, leading to reduced GPX4 ubiquitination. Further research revealed that SAB treatment enhanced the cerebral protective effects of THBS1 inhibition. In conclusion, this study explored the role of exosome-mediated signaling between macrophages and cerebral vascular endothelial cells in CIRI, highlighting the THBS1-OTUD5-GPX4 axis as a driver of endothelial Ferroptosis and brain injury. Targeting this signaling axis represents a potential therapeutic strategy for treating CIRI.

Keywords

Cerebral ischemia–reperfusion injury; Endothelial cells; Exosomes; Ischemic stroke; Macrophage.

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