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Endothelial Cell Derived MVs and Exosomes: Release and Functional Study

Liu, Langni
http://orcid.org/0000-0001-8069-6711
http://rave.ohiolink.edu/etdc/view?acc_num=wright1441015423

Abstract Details

2015, Master of Science (MS), Wright State University, Pharmacology and Toxicology.
Extracellular vesicles are membrane derived vesicles exists in all organs. The two major functioning extracellular vesicles are exosomes (EXMs) and microvesicles (MVs). EXMs released from cell membranes via exocytosis with size between 30-150 nm in diameter and MVs directly budded from plasma membrane and size between 100-1000 nm. Ceramide is one of the most important factor in lipid mechanism for EXM release and also associated with MV release. Ceramide generated from breakdown of sphingomyelin by neutral sphingomyelinase (nSMase). In this study, we used an nSMase inhibitor (GW4869) and an analogue of ceramide (C6-ceramide) to modulate the release of MVs and EXMs in human brain microvascular endothelial cells (ECs), and to test the effects of these modulations on EC proliferation, microRNA126 (miR-126) expression and MV uptake ability and the interaction between EC derived MVs/EXMs (EC-MVs/EXMs) with target cell, human brain smooth muscle cells (SMCs). Endothelial cells were treated with GW4869 in 10 µM and C6-ceramide in 10 µM for 24 h. MVs were isolated from by centrifuge and EXMs were isolated by centrifuge at 20,000 x g for 2 h and EXM were isolated by ultracentrifuge at 120,000 for 2 h with 100 nm filtration. MVs and EXMs size and concentration were analyzed by Nanoparticle Tracking Analysis (NTA) technique. Cell proliferation was detected by MTT. The levels of miRNA126 expression in ECs and EC-MVS/EXMs were detected by qRT-PCR. MV uptake measured by fluorescent microscope. Migration measured by wound healing method. GW4869 significantly decreased EXM release and C6-ceramide significantly increased both MV and EXM release in ECs. ECs treated with C6-ceramide and GW4869 had different miR-126 expression and MV uptake ability. Modulated EC-MVs/EXMs had no effect on normal SMC proliferation and migration, but EC-EXMs under both release regulation decreased angiotensin II (Ang II) induced SMC migration independent of miR-126 level in EXMs. Our data demonstrate that modulation of MV/EXM via C6-ceramide and GW4869 could affects EC function and it`s derived MVs/EXMs function, which may imply in vascular physiology and pathology.
Yanfang Chen, Ph.D. (Advisor)
Ji Chen Bihl, Ph.D. (Committee Member)
Norma Adragna, Ph.D. (Committee Member)
52 p.
Biomedical Research; Pharmacology
microvesicle, exosome, release study

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Citations

  • Liu, L. (2015). Endothelial Cell Derived MVs and Exosomes: Release and Functional Study [Master's thesis, Wright State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=wright1441015423

    APA Style (7th edition)

  • Liu, Langni. Endothelial Cell Derived MVs and Exosomes: Release and Functional Study . 2015. Wright State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=wright1441015423.

    MLA Style (8th edition)

  • Liu, Langni. "Endothelial Cell Derived MVs and Exosomes: Release and Functional Study ." Master's thesis, Wright State University, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=wright1441015423

    Chicago Manual of Style (17th edition)

wright1441015423
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This open access ETD is published by Wright State University and OhioLINK.