hUMSC-derived exosomes alleviate hypoxic cerebrovascular injury via AMPK/NLRP3-mediated pyroptosis suppression and mitochondrial protection

Abstract

As the most prevalent cause of death worldwide, ischemic stroke urgently requires innovative therapeutic strategies. The present study demonstrated the therapeutic potential of human umbilical cord-derived mesenchymal stem cell-derived exosomes (hUMSC-Exos) in ameliorating hypoxia-induced cerebrovascular endothelial dysfunction through modulation of the AMPK/NLRP3 signaling pathway. Bioinformatics analysis of DisGeNET and exosomal cargo databases revealed 283 overlapping cerebral ischemia-related genes, implicating hUMSC-Exos in inflammatory regulation. In vitro experiments showed that hUMSC-Exos rescued oxygen-glucose deprivation (OGD)-induced endothelial dysfunction in bEnd.3 mouse brain endothelial cells, restoring viability, migration, and mitochondrial integrity. Mechanistically, hUMSC-Exos reversed OGD-induced AMPK inactivation while suppressing NLRP3 inflammasome activation, caspase-1 cleavage, and gasdermin D (GSDMD)-mediated pyroptosis. Molecular docking revealed that DL-3-n-butylphthalide acts as a dual-target ligand for AMPK/NLRP3, synergizing with hUMSC-Exos to enhance endothelial protection. In vivo, combined therapy in the transient middle cerebral artery occlusion mouse model reduced cerebral infarction and improved neurological outcomes, accompanied by NLRP3/GSDMD downregulation and hippocampal neuron preservation. These findings establish hUMSC-Exos as regulators of AMPK/NLRP3-mediated pyroptosis and propose a translatable combinatorial regimen for ischemic stroke therapy.

Keywords

stroke / hUMSC-Exos / AMPK/NLRP3 signaling pathway / pyroptosis / synergistic combination therapy

Funding

This work was supported by the National Key Research and Development Program of China (Grant No. 2022YFE0108600 to Y.M.L.), the National Natural Science Foundation of China (Grant Nos. 82460709 and 81960663 to Q.B.L.), the Key Project of Hainan Provincial Department of Education (Grant No. Hnky2019ZD-25 to Q.B.L.), and the Hainan Province High-level Talent Project (Grant No. 2019RC228 to Q.B.L.).

Acknowledgments

None.

Additional information

The online version contains supplementary materials available at http://www.jbr-pub.org.cn/article/doi/10.7555/JBR.39.20250189?pageType=en.

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