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SLC15A3 is transcriptionally regulated by HIF1α and p65 to worsen neuroinflammation in experimental ischemic stroke

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Abstract

Systemic inflammatory stimulus is a risk factor for the incidence of ischemic stroke and contributes to poorer clinical outcomes. Solute carrier 15A3 (SLC15A3) is a peptide/histidine transporter that is implicated in regulating inflammatory responses. However, whether SLC15A3 affects the progression of ischemic stroke associated with systemic inflammation is unclear. The transient middle cerebral artery occlusion (tMCAO) mice with LPS administration (LPS/tMCAO) were prepared as an in vivo model, and LPS-induced BV2 cells under oxygen–glucose deprivation (OGD) exposure were utilized as an in vitro model. We found that SLC15A3 was highly expressed in the ischemic penumbra of LPS/tMCAO mice, and its inhibition reduced infarct area, attenuated neurological deficit, recovered motor function, and mitigated apoptotic neurons. Knockdown of SLC15A3 suppressed the proinflammatory M1-type markers and promoted the levels of M2-associated genes. The in vitro results confirmed that SLC15A3 overexpression promoted microglia polarizing towards M1 subtypes, while SLC15A3 inhibition exerted an opposite effect. In addition, we demonstrated that the p65 signaling pathway and HIF1α were activated by LPS/OGD. Luciferase reporter assay showed that inhibiting p65 using its specific inhibitor BAY 11–7082 or silencing HIF1α using siRNAs reduced the transcriptional activity of SLC15A3 in LPS/OGD-induced BV2 cells. Results in NIH 3T3 cells also confirmed that p65 and HIF1α directly bound to the SLC15A3 promoter to activate SLC15A3 transcription. In conclusion, this work shows that SLC15A3, transcriptionally activated by p65 and HIF1α, contributes to poor outcomes in ischemic stroke associated with systemic inflammation by promoting microglial cells polarizing towards M1 types.

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Data availability

The data are available from the corresponding author on reasonable request.

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Acknowledgements

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Funding

This study was supported by the Youth Fund Project of National Natural Science Foundation of China (82001226) and the Natural Science Foundation of Jilin Province (20210101357JC).

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  1. Shan Yu and Jinghui Yang contributed equally to this work.

Authors and Affiliations

  1. Department of Neurology, China-Japan Union Hospital of Jilin University, No. 126, Xiantai Street, Changchun, 130033, Jilin, China

    Shan Yu, Rui Zhang, Qian Guo & Lu Wang

  2. Department of Hepatobiliary and Pancreatic Surgery, China-Japan Union Hospital of Jilin University, Changchun, 130033, Jilin, China

    Jinghui Yang

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  1. Shan Yu
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Contributions

SY and JHY designed research; SY, RZ, and JHY performed the research; SY, QG and LW analyzed data; SY and JHY wrote the paper. All authors read and approved the final paper.

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Correspondence to Shan Yu.

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The animal experiments were performed complied with the Guide for the Care and Use of Laboratory Animals, and the ethics were approved by the ethical committee of Jilin University (No. 202002031).

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Yu, S., Yang, J., Zhang, R. et al. SLC15A3 is transcriptionally regulated by HIF1α and p65 to worsen neuroinflammation in experimental ischemic stroke. Mol Neurobiol (2024). https://doi.org/10.1007/s12035-024-04191-8

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