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Upregulation of girdin delays endothelial cell apoptosis via promoting engulfment of platelets

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Abstract

Background

Endothelial cells are crucial in maintaining the homeostasis of the blood-brain barrier. Girders of actin filament (Girdin) and phosphor (p)-Girdin are essential for the engulfment of human brain microvascular endothelial cells (HBMECs) into platelets (PLTs), but the potential mechanism remains unclear and requires further study.

Methods

Following PLT and cytochalasin D treatment, Hoechst 33,342 detected apoptosis. The transfection efficiency of the short hairpin RNA targeting Girdin (sh-Girdin) or overexpressing Girdin (OE-Girdin) was determined using western blotting. Sh-Girdin, OE-Girdin, mutated Girdin (m-Girdin), and microfilament binding region deleted Girdin (Del-Girdin) were transfected into HBMECs under PLT conditions. Subsequently, the engulfment of HBMECs by PLTs was detected by flow cytometry and transmission electron microscopy. Girdin and phosphorylated (p)-Girdin levels were quantified by western blot. The positive expression of Girdin was measured by immunohistochemistry (IHC). The localization of PLT, Girdin, and p-Girdin and the engulfment of HBMECs in PLTs were analyzed by confocal microscopy.

Result

Cytochalasin D overturned the inhibitory effect of PLT on cell apoptosis. OE-Girdin enhanced the fluorescent intensity of PLT-labelling and the engulfment of HBMECs by PLTs, while sh-Girdin, m-Girdin, and Del-Girdin ran reversely. OE-Girdin elevated the Girdin and p-Girdin levels, while sh-Girdin and Del-Girdin were the opposite, but m-Girdin did not affect the p-Girdin and Girdin levels.

Conclusion

Girdin and p-Girdin were co-located with PLTs in HBMECs. The over-expression of Girdin was identified as being associated with the increasing engulfment of PTLs. Girdin may be an effective target to alleviate endothelial cell apoptosis.

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Data Availability

Data will be made available on reasonable request.

References

  1. Knox EG, Aburto MR, Clarke G, Cryan JF, O’Driscoll CM (2022) The blood-brain barrier in aging and neurodegeneration. Mol Psychiatry 27(6):2659–2673

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Teleanu RI, Preda MD, Niculescu AG, Vladâcenco O, Radu CI, Grumezescu AM, Teleanu DM (2022) Current strategies to enhance delivery of drugs across the blood–brain barrier. Pharmaceutics 14(5):987

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Trimm E, Red-Horse K (2023) Vascular endothelial cell development and diversity. Nat Reviews Cardiol 20(3):197–210

    Article  Google Scholar 

  4. Sturtzel C (2017) Endothelial cells. Adv Exp Med Biol 1003:71–91

    Article  CAS  PubMed  Google Scholar 

  5. Jia G, Aroor AR, Jia C et al (2019) Endothelial cell senescence in aging-related vascular dysfunction. Biochim et Biophys acta Mol basis disease 1865(7):1802–1809

    Article  CAS  Google Scholar 

  6. Sepúlveda C, Palomo I, Fuentes E (2017) Mechanisms of endothelial dysfunction during aging: predisposition to thrombosis. Mech Ageing Dev 164:91–99

    Article  PubMed  Google Scholar 

  7. Mussbacher M, Schossleitner K, Kral-Pointner JB, Salzmann M, Schrammel A, Schmid JA (2022) More than just a monolayer: the multifaceted role of endothelial cells in the pathophysiology of atherosclerosis. Curr Atheroscler Rep 24(6):483–492

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Aron L, Zullo J, Yankner BA (2022) The adaptive aging brain. Curr Opin Neurobiol 72:91–100

    Article  CAS  PubMed  Google Scholar 

  9. Shevyrev D, Tereshchenko V, Berezina TN, Rybtsov S (2023) Hematopoietic stem cells and the Immune System in Development and Aging. Int J Mol Sci 24(6):5862

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Blacher E, Tsai C, Litichevskiy L, Shipony Z, Iweka CA, Schneider KM, Chuluun B, Heller HC, Menon V, Thaiss CA, Andreasson KI (2022 Feb) Aging disrupts circadian gene regulation and function in macrophages. Nat Immunol 23(2):229–236

  11. Peradinovic J, Mohovic N, Bulic K, Markovinovic A, Cimbro R, Munitic I (2023) Ageing-Induced decline in primary myeloid cell phagocytosis is unaffected by optineurin insufficiency. Biology 12(2):240

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Duan W, Jin Y, Cui Y, Xi F, Liu X, Wo F, Wu J (2021) A co-delivery platform for synergistic promotion of angiogenesis based on biodegradable, therapeutic and self-reporting luminescent porous silicon microparticles. Biomaterials 272:120772

    Article  CAS  PubMed  Google Scholar 

  13. Catan A, Turpin C, Diotel N et al (2019) Aging and glycation promote erythrocyte phagocytosis by human endothelial cells: potential impact in atherothrombosis under diabetic conditions. Atherosclerosis 291:87–98

    Article  CAS  PubMed  Google Scholar 

  14. Kuckleburg CJ, McClenahan DJ, Czuprynski CJ (2008) Platelet activation by Histophilus somni and its lipooligosaccharide induces endothelial cell proinflammatory responses and platelet internalization. Shock (Augusta Ga) 29(2):189–196

    Article  CAS  PubMed  Google Scholar 

  15. Burlak C, Paris LL, Chihara RK et al (2010) The fate of human platelets perfused through the pig liver: implications for xenotransplantation. Xenotransplantation 17(5):350–361

    Article  PubMed  Google Scholar 

  16. Peng Q, Yeh H, Wei L et al (2012) Mechanisms of xenogeneic baboon platelet aggregation and phagocytosis by porcine liver sinusoidal endothelial cells. PLoS ONE 7(10):e47273

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Seta Y, Kawakatsu K, Degawa S, Goto T, Nishikata T (2023) Morphological evidence for Novel Roles of Microtubules in Macrophage phagocytosis. Int J Mol Sci 24(2):1373

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Paone S, Olivieri A (2022) Role of host small GTPases in Apicomplexan Parasite infection. Microorganisms 10(7):1370

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Zhang Y, Liu X, Zhao J, Wang J, Song Q, Zhao C (2022) The phagocytic receptors of β-glucan. Int J Biol Macromol 205:430–441

    Article  CAS  PubMed  Google Scholar 

  20. Lalnunthangi A, Dakpa G, Tiwari S (2023) Multifunctional role of the ubiquitin proteasome pathway in phagocytosis. Prog Mol Biol Transl Sci 194:179–217

    Article  PubMed  Google Scholar 

  21. Cheng SY, Chen NF, Lin PY, Su JH, Chen BH, Kuo HM, Sung CS, Sung PJ, Wen ZH, Chen WF (2019) Anti-invasion and antiangiogenic effects of stellettin B through inhibition of the Akt/Girdin signaling pathway and VEGF in glioblastoma cells. Cancers 11(2):220

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Pan S, Ren F, Li L, Liu D, Li Y, Wang A, Li W, Dong Y, Guo W (2020) MiR-328-3p inhibits cell proliferation and metastasis in colorectal cancer by targeting Girdin and inhibiting the PI3K/Akt signaling pathway. Exp Cell Res 390(1):111939

    Article  CAS  PubMed  Google Scholar 

  23. Wu L, Hua Y, Chang F, Kong D (2019) Girdin: an essential component of pre-replicative complex in human cells. bioRxiv. :634626

  24. Lan Y, Li Y, Li D et al (2018) Engulfment of platelets delays endothelial cell aging via girdin and its phosphorylation. Int J Mol Med 42(2):988–997

    CAS  PubMed  Google Scholar 

  25. Jiang P, Ren YL, Lan Y, Maywood et al (2015) NJ) 240(7):876–883

    CAS  Google Scholar 

  26. Wang W, Chen H, Gao W et al (2020) Girdin interaction with vimentin induces EMT and promotes the growth and metastasis of pancreatic ductal adenocarcinoma. Oncol Rep 44(2):637–649

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Cortes-Canteli M, Iadecola C (2020) Alzheimer’s Disease and Vascular Aging: JACC Focus Seminar. J Am Coll Cardiol 75(8):942–951

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Li G, Yang T, Liu Y, Su H, Liu W, Fang D, Jin L, Jin F, Xu T, Duan C (2023) The proteins derived from platelet-rich plasma improve the endothelialization and vascularization of small diameter vascular grafts. Int J Biol Macromol 225:574–587

    Article  CAS  PubMed  Google Scholar 

  29. Fujita M, Horio T, Kishimoto S et al (2013) Effects of platelet-rich plasma-containing fragmin/protamine microparticles in enhancing endothelial and smooth muscle cell growth and inducing collateral vessels in a rabbit model of hindlimb ischemia. J biomedical Mater Res Part B Appl biomaterials 101(1):36–42

    Article  Google Scholar 

  30. Ranzato E, Boccafoschi F, Mazzucco L et al (2010) Role of ERK1/2 in platelet lysate-driven endothelial cell repair. J Cell Biochem 110(3):783–793

    Article  CAS  PubMed  Google Scholar 

  31. Cho MS, Bottsford-Miller J, Vasquez HG et al (2012) Platelets increase the proliferation of ovarian cancer cells. Blood 120(24):4869–4872

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Lang D, Dohle F, Terstesse M et al (1950) Down-regulation of monocyte apoptosis by phagocytosis of platelets: involvement of a caspase-9, caspase-3, and heat shock protein 70-dependent pathway. Journal of immunology (Baltimore, Md: 2002,168(12):6152-8

  33. Ji S, Dong W, Qi Y et al (2020) Phagocytosis by endothelial cells inhibits procoagulant activity of platelets of essential thrombocythemia in vitro. J Thromb haemostasis: JTH 18(1):222–233

    Article  CAS  Google Scholar 

  34. Takahashi M, Asai N, Enomoto A (2015) Akt-girdin as oncotarget. Oncoscience 2(10):811–812

    Article  PubMed  PubMed Central  Google Scholar 

  35. Miyake H, Maeda K, Asai N et al (2011) The actin-binding protein Girdin and its akt-mediated phosphorylation regulate neointima formation after vascular injury. Circul Res 108(10):1170–1179

    Article  CAS  Google Scholar 

  36. Lansdown DA, Morrison C, Zaid MB et al (2019) Preoperative IDEAL (iterative decomposition of Echoes of asymmetrical length) magnetic resonance imaging rotator cuff muscle fat fractions are associated with rotator cuff repair outcomes. J Shoulder Elbow Surg 28(10):1936–1941

    Article  PubMed  Google Scholar 

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Funding

This work was supported by the National Natural Science Foundation of China no 31371160.

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Author notes

  1. Yong Lan and Min Dong contributed equally as co-first authors.

Authors and Affiliations

  1. Department of Vascular Surgery, Beijing hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing, P.R. China

    Yong Lan, Yongjun Li, Yongpeng Diao, Zuoguan Chen & Zhiyuan Wu

  2. Department of Cardiology, Beijing hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing, P.R. China

    Min Dong

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  1. Yong Lan
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Correspondence to Yong Lan.

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Lan, Y., Dong, M., Li, Y. et al. Upregulation of girdin delays endothelial cell apoptosis via promoting engulfment of platelets. Mol Biol Rep 50, 8111–8120 (2023). https://doi.org/10.1007/s11033-023-08625-9

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