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MMP-9 inhibitor SB-3CT improves neurological outcomes in ischemic stroke mice by modulation of astrocytic lipid metabolism

Acta Pharmacologica Sinica volume 46, pages 2120–2135 (2025)Cite this article

Abstract

The acute phase of ischemic stroke is marked by a surge in matrix metalloproteinase-9 (MMP-9) activity. While integral to natural repair processes, MMP-9 exacerbates injury by breaking down the blood-brain barrier (BBB) and promoting edema and inflammation. MMP-9 is predominantly secreted by inflammatory cells such as neutrophils, macrophages and microglia soon after stroke onset. In this study we investigated the effects of MMP-9 inhibition via SB-3CT on astrocytic lipid metabolism, and its potential to enhance neuronal survival and recovery following ischemic stroke. Mice were subjected to transient middle cerebral artery occlusion (tMCAO) for 60 min, mice then were injected with SB-3CT (25 mg/kg, i.v.). On D3 post tMCAO, neurological outcomes were assessed, and whole brains were collected for analysis. Lipidomic analysis of brain tissue showed that SB-3CT treatment significantly restrained astrocytic cholesterol metabolism by modulating the sphingolipid and glycerophospholipid pathways. Specifically, SB-3CT reduced ceramide accumulation and promoted an increase in neuroprotective hexosylceramides, leading to enhanced neuronal survival and synaptic integrity. In addition, SB-3CT treatment reduced astrocytic and microglial reactivity, thereby mitigating neuroinflammation. In order to optimize the timing and dosage of MMP-9 inhibition to maximize the therapeutic efficacy, tMCAO mice were given three injections of SB-3CT on D0, D2 and D4 within 7 days after modeling. We found that prolonged MMP-9 inhibition alleviated astrogliosis, concurrently impaired neurological recovery and inhibited angiogenesis. These results demonstrate the critical role of lipid metabolism in MMP-9-mediated brain injury and the potential of SB-3CT as a therapeutic strategy for ischemic stroke by targeting astrocytic lipid metabolism.

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Fig. 1: Lipidomic profiling reveals lipid metabolism changes following MMP-9 inhibition and neuroprotection.
Fig. 2: Modulation of glycerophospholipid and cholesterol metabolism by SB-3CT enhances neuronal recovery post-ischemia.
Fig. 3: MMP-9 inhibition mediates SB-3CT’s regulatory effects on cholesterol metabolism and astrocyte reactivity.
Fig. 4: Enhanced neurological recovery and neurogenesis post-ischemic event via MMP-9 inhibition by SB-3CT.
Fig. 5: MMP-9 inhibition enhances neurological function recovery after cerebral ischemia-reperfusion injury.
Fig. 6: Continuous MMP-9 inhibition: balancing neurological recovery and potential detrimental effects.

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Data availability

The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We would like to express our gratitude to Climb Technology Co., Ltd and Dr. Min Yao for their assistance with lipidomics analysis in our study.

Funding

This work was supported by the National Natural Science Foundation of China (31900704 to CF, 82273923 to YZM), Shandong Provincial Key Innovation Program (2019JZZY020909 to LDD) Guangdong Province Basic and Applied Basic Research Grant (2023A1515010467 to CF), Shenzhen Science and Technology Program (JCYJ20220531100203008 to YZM, JCYJ20230807140721043 to CF). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Author notes

  1. These authors contributed equally: Li-da Du, Cheng Fang, Yue-qing Wang

  2. These authors jointly supervised this work: Ju-yang Huang, Yin-zhong Ma

Authors and Affiliations

  1. Institute of Molecular Medicine & Innovative Pharmaceutics, Qingdao University, Qingdao, 266071, China

    Li-da Du

  2. Provincial Laboratory of Polymorphic Medicine, Tengzhou, 277599, China

    Li-da Du & Zhao-lin Gao

  3. Institute of Biomedicine and Biotechnology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China

    Cheng Fang, Yue-qing Wang, Zi-ying Feng, Ogunleye Femi Abiola & Yin-zhong Ma

  4. University of Chinese Academy of Sciences, Beijing, 101408, China

    Yue-qing Wang

  5. School of Pharmaceutical Sciences (Shenzhen). Sun Yat-sen University, Shenzhen, 518107, China

    Ju-yang Huang

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Contributions

YZM: Conceived and designed the research; supervised the project; contributed to data analysis and interpretation; wrote the manuscript. JYH: Contributed to experimental design; provided new reagents and resources; supervised lipidomic analysis. LDD: Performed animal surgeries and behavioral assessments; conducted immunofluorescence staining experiments; contributed to data collection. CF: Contributed to the design of lipidomic experiments; analyzed lipidomic data. ZYF: Assisted with experimental procedures and data collection; contributed to manuscript revision. YQW: Assisted with statistical analysis and interpretation of neurological recovery data. OFA: Conducted tissue collection and Western blot analysis; contributed to data interpretation. ZLG: Reviewed and commented on the manuscript; contributed to manuscript editing.

Corresponding authors

Correspondence to Ju-yang Huang or Yin-zhong Ma.

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The authors declare no competing interests.

Ethical approval

The experimental procedures performed on animals in this study were strictly in accordance with the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996), and approved by the Institutional Animal Care and Use Committee (IACUC) of Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences. All efforts were made to minimize animal suffering and to reduce the number of animals used.

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Du, Ld., Fang, C., Wang, Yq. et al. MMP-9 inhibitor SB-3CT improves neurological outcomes in ischemic stroke mice by modulation of astrocytic lipid metabolism. Acta Pharmacol Sin 46, 2120–2135 (2025). https://doi.org/10.1038/s41401-025-01505-x

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