Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Advertisement

Scientific Reports
  • View all journals
  • Search
  • My Account Login
  • Content Explore content
  • About the journal
  • Publish with us
  • Sign up for alerts
  • RSS feed
  1. nature
  2. scientific reports
  3. articles
  4. article
Brain-derived microvesicles induce activation of aspirin-treated platelets via the PLC/PKC pathway
Download PDF
Download PDF
  • Article
  • Open access
  • Published: 25 March 2026

Brain-derived microvesicles induce activation of aspirin-treated platelets via the PLC/PKC pathway

  • Yi-fan He1,
  • Jia-cheng Zhang2,
  • Yuan-zhi Wang2,
  • Xiao-chun Li2,
  • Rui-ting Zhao4,
  • Yi-dan Gou2,
  • Heng-jie Yuan3 &
  • …
  • Ye Tian2 

Scientific Reports , Article number:  (2026) Cite this article

  • 1586 Accesses

  • Metrics details

We are providing an unedited version of this manuscript to give early access to its findings. Before final publication, the manuscript will undergo further editing. Please note there may be errors present which affect the content, and all legal disclaimers apply.

Subjects

  • Cell biology
  • Diseases
  • Drug discovery
  • Neurology
  • Neuroscience

Abstract

Aspirin (Asp) therapy reduces the risk of arterial thrombotic events. However, aspirin-related adverse effects, particularly cerebral hemorrhage with its high mortality and disability rates, remain a significant concern. Interestingly, long-term antiplatelet therapy does not appear to exacerbate bleeding severity in patients with hemorrhagic cerebrovascular disease, suggesting potential compensatory mechanisms inducing platelet activation post-hemorrhage. Brain-derived microvesicles (BDMVs) have been implicated in platelet activation, although the underlying molecular mechanisms are unclear. This study employed flow cytometry (FCM), ELISA, and hopping probe ion conductance microscopy (HPICM) to demonstrate that BDMVs significantly activate and induce morphological changes in aspirin-treated platelets. We discovered the presence of cyclooxygenase-1 (COX-1) on BDMVs. Then, the phosphorylated proteomics was used to analyze the effect of BDMVs on aspirin-treated platelet quantitatively and validate the involvement of several signaling molecules. Biological validation showed that BDMVs increased phospholipase C (PLC), protein kinase C (PKC) and Akt phosphorylation. We also used PLC inhibitor U73122 to treat BDMV-intervened platelet and found reduced phosphorylation of the downstream signaling molecule PKC. These findings suggest that COX-1 within BDMVs may partially counteract the inhibitory effect of aspirin on platelets. Furthermore, BDMVs, combined with arachidonic acid (AA), activate aspirin-treated platelets and suggest the involvement of the PLC/PKC pathway. This study provides a theoretical basis for the early treatment of patients with clinical aspirin-related cerebral hemorrhage.

Data availability

All data generated or analyzed during this study are included in this published article.

References

  1. Assadian, A. et al. Aspirin resistance among long-term aspirin users after carotid endarterectomy and controls: flow cytometric measurement of aspirin-induced platelet Inhibition. J. Vasc. Surg. 45, 1142–1147. https://doi.org/10.1016/j.jvs.2007.01.064 (2007). discussion 1147.

    Google Scholar 

  2. Roffi, M. et al. ESC Guidelines for the management of acute coronary syndromes in patients presenting without persistent ST-segment elevation: Task Force for the Management of Acute Coronary Syndromes in Patients Presenting without Persistent ST-Segment Elevation of the European Society of Cardiology (ESC). Eur. Heart J. 37, 267–315. https://doi.org/10.1093/eurheartj/ehv320 (2016).

  3. Global, regional, and national burden of stroke and its risk factors, 1990–2019: a systematic analysis for the global burden of disease study 2019. Lancet Neurol. 20, 795–820. https://doi.org/10.1016/s1474-4422(21)00252-0 (2021).

    Google Scholar 

  4. Gross, B. A., Jankowitz, B. T. & Friedlander, R. M. Cerebral intraparenchymal hemorrhage: A review. Jama 321, 1295–1303. https://doi.org/10.1001/jama.2019.2413 (2019).

    Google Scholar 

  5. Hostettler, I. C., Seiffge, D. J. & Werring, D. J. Intracerebral hemorrhage: an update on diagnosis and treatment. Expert Rev. Neurother. 19, 679–694. https://doi.org/10.1080/14737175.2019.1623671 (2019).

    Google Scholar 

  6. Ivascu, F. A. et al. Predictors of mortality in trauma patients with intracranial hemorrhage on preinjury aspirin or clopidogrel. J. Trauma. 65, 785–788. https://doi.org/10.1097/TA.0b013e3181848caa (2008).

    Google Scholar 

  7. Nishijima, D. K., Zehtabchi, S., Berrong, J. & Legome, E. Utility of platelet transfusion in adult patients with traumatic intracranial hemorrhage and preinjury antiplatelet use: a systematic review. J. Trauma. Acute Care Surg. 72, 1658–1663. https://doi.org/10.1097/TA.0b013e318256dfc5 (2012).

    Google Scholar 

  8. Sansing, L. H. et al. Prior antiplatelet use does not affect hemorrhage growth or outcome after ICH. Neurology 72, 1397–1402. https://doi.org/10.1212/01.wnl.0000342709.31341.88 (2009).

    Google Scholar 

  9. Zhou, L. et al. Microparticles: new light shed on the Understanding of venous thromboembolism. Acta Pharmacol. Sin. 35, 1103–1110. https://doi.org/10.1038/aps.2014.73 (2014).

    Google Scholar 

  10. Schiro, A. et al. Endothelial microparticles as conveyors of information in atherosclerotic disease. Atherosclerosis 234, 295–302. https://doi.org/10.1016/j.atherosclerosis.2014.03.019 (2014).

    Google Scholar 

  11. Geddings, J. E. & Mackman, N. Tumor-derived tissue factor-positive microparticles and venous thrombosis in cancer patients. Blood 122, 1873–1880. https://doi.org/10.1182/blood-2013-04-460139 (2013).

    Google Scholar 

  12. Kaptan, K., Beyan, C., Ifran, A. & Pekel, A. Platelet-derived microparticle levels in women with recurrent spontaneous abortion. Int. J. Gynaecol. Obstet. 102, 271–274. https://doi.org/10.1016/j.ijgo.2008.04.007 (2008).

    Google Scholar 

  13. Park, M. S. et al. Quantification of hypercoagulable state after blunt trauma: microparticle and thrombin generation are increased relative to injury severity, while standard markers are not. Surgery 151, 831–836. https://doi.org/10.1016/j.surg.2011.12.022 (2012).

    Google Scholar 

  14. Booth, A. M. et al. Exosomes and HIV gag bud from endosome-like domains of the T cell plasma membrane. J. Cell Biol. 172, 923–935. https://doi.org/10.1083/jcb.200508014 (2006).

    Google Scholar 

  15. Laso-García, F. et al. Protein content of blood-derived extracellular vesicles: an approach to the pathophysiology of cerebral hemorrhage. Front. Cell. Neurosci. 16, 1058546. https://doi.org/10.3389/fncel.2022.1058546 (2022).

    Google Scholar 

  16. van Niel, G., D’Angelo, G. & Raposo, G. Shedding light on the cell biology of extracellular vesicles. Nat. Rev. Mol. Cell Biol. 19, 213–228. https://doi.org/10.1038/nrm.2017.125 (2018).

    Google Scholar 

  17. Urabe, F. et al. Extracellular vesicles as biomarkers and therapeutic targets for cancer. Am. J. Physiol. Cell Physiol. 318, C29–c39. https://doi.org/10.1152/ajpcell.00280.2019 (2020).

    Google Scholar 

  18. Ratajczak, M. Z. & Ratajczak, J. Extracellular microvesicles/exosomes: discovery, disbelief, acceptance, and the future? Leukemia 34, 3126–3135. https://doi.org/10.1038/s41375-020-01041-z (2020).

    Google Scholar 

  19. Alkhamis, T. M., Beissinger, R. L. & Chediak, J. R. Artificial surface effect on red blood cells and platelets in laminar shear flow. Blood 75, 1568–1575 (1990).

    Google Scholar 

  20. Schwager, S. C. & Reinhart-King, C. A. Mechanobiology of microvesicle release, uptake, and microvesicle-mediated activation. Curr. Top. Membr. 86, 255–278. https://doi.org/10.1016/bs.ctm.2020.08.004 (2020).

    Google Scholar 

  21. Raposo, G. & Stoorvogel, W. Extracellular vesicles: exosomes, microvesicles, and friends. J. Cell Biol. 200, 373–383. https://doi.org/10.1083/jcb.201211138 (2013).

    Google Scholar 

  22. Chen, Y. T., Yuan, H. X., Ou, Z. J. & Ou, J. S. Microparticles (Exosomes) and atherosclerosis. Curr. Atheroscler. Rep. 22, 23. https://doi.org/10.1007/s11883-020-00841-z (2020).

    Google Scholar 

  23. Porro, C., Trotta, T. & Panaro, M. A. Microvesicles in the brain: Biomarker, messenger or mediator? J. Neuroimmunol. 288, 70–78. https://doi.org/10.1016/j.jneuroim.2015.09.006 (2015).

    Google Scholar 

  24. Tian, Y. et al. Brain-derived microparticles induce systemic coagulation in a murine model of traumatic brain injury. Blood 125, 2151–2159. https://doi.org/10.1182/blood-2014-09-598805 (2015).

    Google Scholar 

  25. Zhao, Z. et al. Extracellular mitochondria released from traumatized brains induced platelet procoagulant activity. Haematologica 105, 209–217. https://doi.org/10.3324/haematol.2018.214932 (2020).

    Google Scholar 

  26. Leitner, A. Enrichment strategies in phosphoproteomics. Methods Mol. Biol. (Clifton N J). 1355, 105–121. https://doi.org/10.1007/978-1-4939-3049-4_7 (2016).

    Google Scholar 

  27. Zahedi, R. P., Begonja, A. J., Gambaryan, S. & Sickmann, A. Phosphoproteomics of human platelets: A quest for novel activation pathways. Biochim. Biophys. Acta. 1764, 1963–1976. https://doi.org/10.1016/j.bbapap.2006.08.017 (2006).

    Google Scholar 

  28. Zhao, R. T. et al. Circular ribonucleic acid expression alteration in exosomes from the brain extracellular space after traumatic brain injury in mice. J. Neurotrauma. 35, 2056–2066. https://doi.org/10.1089/neu.2017.5502 (2018).

    Google Scholar 

  29. Michelson, A. D. Flow cytometry: a clinical test of platelet function. Blood 87, 4925–4936 (1996).

    Google Scholar 

  30. McEver, R. P. Selectin-carbohydrate interactions during inflammation and metastasis. Glycoconj. J. 14, 585–591. https://doi.org/10.1023/a:1018584425879 (1997).

    Google Scholar 

  31. Serebruany, V. L., McKenzie, M. E., Levin, D. J. & Gurbel, P. A. Monitoring platelet Inhibition during chronic oral platelet glycoprotein IIb/IIIa blockade: are we missing something? Thromb. Haemost. 83, 356–357 (2000).

    Google Scholar 

  32. Liu, X. et al. Use of non-contact hopping probe ion conductance microscopy to investigate dynamic morphology of live platelets. Platelets 26, 480–485. https://doi.org/10.3109/09537104.2014.940888 (2015).

    Google Scholar 

  33. Cox, J. & Mann, M. MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat. Biotechnol. 26, 1367–1372. https://doi.org/10.1038/nbt.1511 (2008).

    Google Scholar 

  34. Cox, J. et al. Accurate proteome-wide label-free quantification by delayed normalization and maximal peptide ratio extraction, termed MaxLFQ. Mol. Cell. Proteomics: MCP. 13, 2513–2526. https://doi.org/10.1074/mcp.M113.031591 (2014).

    Google Scholar 

  35. Ashburner, M. et al. Gene ontology: tool for the unification of biology. The gene ontology consortium. Nat. Genet. 25, 25–29. https://doi.org/10.1038/75556 (2000).

    Google Scholar 

  36. Cheng, A., Grant, C. E., Noble, W. S. & Bailey, T. L. MoMo: discovery of statistically significant post-translational modification motifs. Bioinf. (Oxford England). 35, 2774–2782. https://doi.org/10.1093/bioinformatics/bty1058 (2019).

    Google Scholar 

  37. Edbauer, D. et al. Identification and characterization of neuronal mitogen-activated protein kinase substrates using a specific phosphomotif antibody. Mol. Cell. Proteomics: MCP. 8, 681–695. https://doi.org/10.1074/mcp.M800233-MCP200 (2009).

    Google Scholar 

  38. Seyfert, U. T., Haubelt, H., Vogt, A. & Hellstern, P. Variables influencing Multiplate(TM) whole blood impedance platelet aggregometry and turbidimetric platelet aggregation in healthy individuals. Platelets 18, 199–206. https://doi.org/10.1080/09537100600944277 (2007).

    Google Scholar 

  39. Melki, I., Tessandier, N., Zufferey, A. & Boilard, E. Platelet microvesicles in health and disease. Platelets 28, 214–221. https://doi.org/10.1080/09537104.2016.1265924 (2017).

    Google Scholar 

  40. Duchez, A. C. et al. Platelet microparticles are internalized in neutrophils via the concerted activity of 12-lipoxygenase and secreted phospholipase A2-IIA. Proc. Natl. Acad. Sci. U.S.A. 112, E3564–3573. https://doi.org/10.1073/pnas.1507905112 (2015).

    Google Scholar 

  41. Vanhoutte, P. M. COX-1 and vascular disease. Clin. Pharmacol. Ther. 86, 212–215. https://doi.org/10.1038/clpt.2009.108 (2009).

    Google Scholar 

  42. Vane, J. R., Bakhle, Y. S. & Botting, R. M. Cyclooxygenases 1 and 2. Annu. Rev. Pharmacol. Toxicol. 38, 97–120. https://doi.org/10.1146/annurev.pharmtox.38.1.97 (1998).

    Google Scholar 

  43. Dec, K. et al. Long-term exposure to fluoride as a factor promoting changes in the expression and activity of cyclooxygenases (COX1 and COX2) in various rat brain structures. Neurotoxicology 74, 81–90. https://doi.org/10.1016/j.neuro.2019.06.001 (2019).

    Google Scholar 

  44. Hankey, G. J. & Eikelboom, J. W. Aspirin resistance. Lancet (London England). 367, 606–617. https://doi.org/10.1016/s0140-6736(06)68040-9 (2006).

    Google Scholar 

  45. Macchi, L., Sorel, N. & Christiaens, L. Aspirin resistance: definitions, mechanisms, prevalence, and clinical significance. Curr. Pharm. Des. 12, 251–258. https://doi.org/10.2174/138161206775193064 (2006).

    Google Scholar 

  46. Khismatullin, R. R. et al. Pathology of lung-specific thrombosis and inflammation in COVID-19. J. Thromb. Haemostasis: JTH. 19, 3062–3072. https://doi.org/10.1111/jth.15532 (2021).

    Google Scholar 

  47. Denorme, F. & Campbell, R. A. Procoagulant platelets: novel players in thromboinflammation. Am. J. Physiol. Cell Physiol. 323, C951–c958. https://doi.org/10.1152/ajpcell.00252.2022 (2022).

    Google Scholar 

  48. Kulkarni, P. P. & McCrae, K. R. Comprehensive analysis of procoagulant platelets exhibiting features of Necrosis, apoptosis and platelet activation. J. Vis. Exp. JoVE. https://doi.org/10.3791/68116 (2025).

    Google Scholar 

  49. Veuthey, L., Aliotta, A., Bertaggia Calderara, D., Pereira Portela, C. & Alberio, L. Mechanisms underlying dichotomous procoagulant COAT platelet Generation-A conceptual review summarizing current knowledge. Int. J. Mol. Sci. 23 https://doi.org/10.3390/ijms23052536 (2022).

  50. Prus, G., Hoegl, A., Weinert, B. T. & Choudhary, C. Analysis and interpretation of protein Post-Translational modification site stoichiometry. Trends Biochem. Sci. 44, 943–960. https://doi.org/10.1016/j.tibs.2019.06.003 (2019).

    Google Scholar 

  51. Cox, J. et al. Andromeda: a peptide search engine integrated into the MaxQuant environment. J. Proteome Res. 10, 1794–1805. https://doi.org/10.1021/pr101065j (2011).

    Google Scholar 

  52. Lee, E. E., Winston-Gray, C., Barlow, J. W., Rissman, R. A. & Jeste, D. V. Plasma levels of neuron- and astrocyte-derived exosomal amyloid Beta1-42, amyloid Beta1-40, and phosphorylated Tau levels in schizophrenia patients and Non-psychiatric comparison subjects: relationships with cognitive functioning and psychopathology. Front. Psychiatry. 11, 532624. https://doi.org/10.3389/fpsyt.2020.532624 (2020).

    Google Scholar 

  53. Küry, S. et al. De novo mutations in protein kinase genes CAMK2A and CAMK2B cause intellectual disability. Am. J. Hum. Genet. 101, 768–788. https://doi.org/10.1016/j.ajhg.2017.10.003 (2017).

    Google Scholar 

  54. Bill, C. A. & Vines, C. M. Phospholipase C. Adv. Exp. Med. Biol. 1131, 215–242. Doi:https://doi.org/10.1007/978-3-030-12457-1_9 (2020).

  55. Stabel, S. & Parker, P. J. Protein kinase C. Pharmacol. Ther. 51, 71–95. https://doi.org/10.1016/0163-7258(91)90042-k (1991).

  56. Moore, D. T., Berger, B. W. & DeGrado, W. F. Protein-protein interactions in the membrane: sequence, structural, and biological motifs. Structure (London, England). 16, 991–1001. https://doi.org/10.1016/j.str.2008.05.007 (2008).

  57. Ardito, F., Giuliani, M., Perrone, D., Troiano, G. & Lo Muzio, L. The crucial role of protein phosphorylation in cell signaling and its use as targeted therapy (Review). Int. J. Mol. Med. 40, 271–280. https://doi.org/10.3892/ijmm.2017.3036 (2017).

    Google Scholar 

  58. Rubenstein, D. A. & Yin, W. Platelet-activation mechanisms and vascular remodeling. Compr. Physiol. 8, 1117–1156. https://doi.org/10.1002/cphy.c170049 (2018).

    Google Scholar 

  59. Li, Z., Delaney, M. K., O’Brien, K. A. & Du, X. Signaling during platelet adhesion and activation. Arterioscler. Thromb. Vasc. Biol. 30, 2341–2349. https://doi.org/10.1161/atvbaha.110.207522 (2010).

    Google Scholar 

Download references

Funding

This study was funded by major project of the Tianjin Natural Science Foundation (Tianjin Municipal Science and Technology Commission) (No. 21ZXJBSY00050) and the National Natural Science Foundation of China (No. 81971173 and 82371386).

Author information

Authors and Affiliations

  1. Department of Pharmacy, Hebei General Hospital, Shijiazhuang, 050051, Hebei, China

    Yi-fan He

  2. Department of Neurosurgery, Tianjin Neurological Institute, Tianjin Medical University General Hospital, Tianjin, 300052, China

    Jia-cheng Zhang, Yuan-zhi Wang, Xiao-chun Li, Yi-dan Gou & Ye Tian

  3. Department of Pharmacy, Tianjin Medical University General Hospital, Tianjin, 300052, China

    Heng-jie Yuan

  4. Department of Pharmacy, Tianjin Medical University General Hospital Airport Hospital, Tianjin, 300308, China

    Rui-ting Zhao

Authors
  1. Yi-fan He
    View author publications

    Search author on:PubMed Google Scholar

  2. Jia-cheng Zhang
    View author publications

    Search author on:PubMed Google Scholar

  3. Yuan-zhi Wang
    View author publications

    Search author on:PubMed Google Scholar

  4. Xiao-chun Li
    View author publications

    Search author on:PubMed Google Scholar

  5. Rui-ting Zhao
    View author publications

    Search author on:PubMed Google Scholar

  6. Yi-dan Gou
    View author publications

    Search author on:PubMed Google Scholar

  7. Heng-jie Yuan
    View author publications

    Search author on:PubMed Google Scholar

  8. Ye Tian
    View author publications

    Search author on:PubMed Google Scholar

Contributions

Y.H. performed the experiments, analyzed the data and wrote the paper; J.Z., Y.W., and X.L. participated in the execution of the experiment; R.Z. and Y.G. contributed to analysis and manuscript preparation; H.Y. and Y.T. contributed to the conception of the study. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Heng-jie Yuan or Ye Tian.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary Material 1 (download PDF )

Supplementary Material 2 (download XLSX )

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

He, Yf., Zhang, Jc., Wang, Yz. et al. Brain-derived microvesicles induce activation of aspirin-treated platelets via the PLC/PKC pathway. Sci Rep (2026). https://doi.org/10.1038/s41598-026-39509-0

Download citation

  • Received: 29 June 2025

  • Accepted: 05 February 2026

  • Published: 25 March 2026

  • DOI: https://doi.org/10.1038/s41598-026-39509-0

Share this article

Anyone you share the following link with will be able to read this content:

Sorry, a shareable link is not currently available for this article.

Provided by the Springer Nature SharedIt content-sharing initiative

Keywords

  • Platelet
  • Brain-derived microvesicles
  • Aspirin
  • Phosphoproteomics
  • PLC/PKC pathway
Download PDF

Advertisement

Explore content

  • Research articles
  • News & Comment
  • Collections
  • Subjects
  • Follow us on Facebook
  • Follow us on X
  • Sign up for alerts
  • RSS feed

About the journal

  • About Scientific Reports
  • Contact
  • Journal policies
  • Guide to referees
  • Calls for Papers
  • Editor's Choice
  • Journal highlights
  • Open Access Fees and Funding

Publish with us

  • For authors
  • Language editing services
  • Open access funding
  • Submit manuscript

Search

Advanced search

Quick links

  • Explore articles by subject
  • Find a job
  • Guide to authors
  • Editorial policies

Scientific Reports (Sci Rep)

ISSN 2045-2322 (online)

nature.com footer links

About Nature Portfolio

  • About us
  • Press releases
  • Press office
  • Contact us

Discover content

  • Journals A-Z
  • Articles by subject
  • protocols.io
  • Nature Index

Publishing policies

  • Nature portfolio policies
  • Open access

Author & Researcher services

  • Reprints & permissions
  • Research data
  • Language editing
  • Scientific editing
  • Nature Masterclasses
  • Research Solutions

Libraries & institutions

  • Librarian service & tools
  • Librarian portal
  • Open research
  • Recommend to library

Advertising & partnerships

  • Advertising
  • Partnerships & Services
  • Media kits
  • Branded content

Professional development

  • Nature Awards
  • Nature Careers
  • Nature Conferences

Regional websites

  • Nature Africa
  • Nature China
  • Nature India
  • Nature Japan
  • Nature Middle East
  • Privacy Policy
  • Use of cookies
  • Legal notice
  • Accessibility statement
  • Terms & Conditions
  • Your US state privacy rights
Springer Nature

© 2026 Springer Nature Limited

Nature Briefing: Translational Research

Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology, drug discovery and pharma.

Get what matters in translational research, free to your inbox weekly. Sign up for Nature Briefing: Translational Research