Advances in Clinical and Experimental Medicine
2019, vol. 28, nr 4, April, p. 421–430
Publication type: original article
IL-4-polarized BV2 microglia cells promote angiogenesis by secreting exosomes
1 Department of Immunology, College of Basic Medical Sciences, Jilin University, Changchun, China
2 Key Laboratory of Molecular Enzymology and Engineering under the Ministry of Education, College of Life Sciences, Jilin University, Changchun, China
3 Department of Neurosurgery, Stanford University, USA
4 Department of Obstetrics, First Hospital of Jilin University, Changchun, China
5 Intensive Care Unit, the Affiliated Hospital of Qingdao University, China
Background. The microglia cell transfer has been shown to play a protective role in ischemic stroke. Microglia cells may play this nerve-protective role via the promotion of angiogenesis. However, the underlying mechanisms are largely unknown and need further investigation.
Objectives. The aim of this study was to investigate the pro-angiogenesis effects of unpolarized, interleukin-4 (IL-4)-polarized or lipopolysaccharide (LPS)-polarized BV2 microglia cells both in vivo and in vitro. We also investigated the potential mechanisms of these pro-angiogenesis effects.
Material and Methods. BV2 cells were polarized using phosphate-buffered saline (PBS), LPS or IL-4, respectively. The gene expression pattern was analyzed by reverse transcription polymerase chain reaction (RTPCR). The transfer of polarized BV2 cells was performed with an intravenous injection into mice 45 min after the middle cerebral artery (MCA) occlusion. Angiogenin expression was assessed by immunofluorescence. We also examined the angiogenesis effect of polarized BV2 cells and their exosomes through 3-dimensional co-cultures in vitro. Finally, the microRNA (miRNA) profiles of exosomes released by BV2 cells under different polarization conditions were examined using miRNA microarray.
Results. The IL-4-polarized BV2 transplantation promoted angiogenin expression in the ischemic brain. Interleukin-4-polarized microglia increased the tube formation of endothelial cells by secreting exosomes. The miRNA profiles of exosomes released by BV2 cells under different polarization conditions were different. Exosomes from IL-4-polarized BV2 cells contained higher amounts of miRNA-26a compared to those from the LPS-polarized and unpolarized BV2 cells.
Conclusion. Interleukin-4-polarized microglia cells might ameliorate the damage caused by ischemic stroke by promoting angiogenesis through the secretion of exosomes containing miRNA-26a.
angiogenesis, exosomes, microglia, interleukin-4
- Fan Y, Xie L, Chung CY. Signaling pathways controlling microglia chemotaxis. Mol Cells. 2017;40(3):163–168.
- Kanazawa M, Miura M, Toriyabe M, et al. Microglia preconditioned by oxygen-glucose deprivation promote functional recovery in ischemic rats. Sci Rep. 2017;7:42582.
- Narantuya D, Nagai A, Sheikh AM, et al. Human microglia transplanted in rat focal ischemia brain induce neuroprotection and behavioral improvement. PLoS ONE. 2010;5(7):e11746.
- Hu X, Li P, Guo Y, et al. Microglia/macrophage polarization dynamics reveal novel mechanism of injury expansion after focal cerebral ischemia. Stroke. 2012;43(11):3063–3070.
- Lee JA, Song HY, Ju SM, et al. Suppression of inducible nitric oxide synthase and cyclooxygenase-2 by cell-permeable superoxide dismutase in lipopolysaccharide-stimulated BV-2 microglial cells. Mol Cells. 2010;29(3):245–250.
- Korhonen P, Kanninen KM, Lehtonen S, et al. Immunomodulation by interleukin-33 is protective in stroke through modulation of inflammation. Brain Behav Immun. 2015;49:322–336.
- Liu X, Liu J, Zhao S, et al. Interleukin-4 is essential for microglia/macrophage M2 polarization and long-term recovery after cerebral ischemia. Stroke. 2016;47(2):498–504.
- Amantea D, Certo M, Petrelli F, et al. Azithromycin protects mice against ischemic stroke injury by promoting macrophage transition towards M2 phenotype. Exp Neurol. 2016;275(Pt 1):116–125.
- Chernykh ER, Shevela EY, Starostina NM, et al. Safety and therapeutic potential of M2-macrophages in stroke treatment. Cell Transplant. 2015;25(8):1461–1471.
- Desestret V, Riou A, Chauveau F, et al. In vitro and in vivo models of cerebral ischemia show discrepancy in therapeutic effects of M2 macrophages. PLOS ONE. 2013;8(6):e67063.
- Fumagalli S, Perego C, Pischiutta F, Zanier ER, De Simoni MG. The ischemic environment drives microglia and macrophage function. Front Neurol. 2015;6:81.
- Xin H, Li Y, Cui Y, Yang JJ, Zhang ZG, Chopp M. Systemic administration of exosomes released from mesenchymal stromal cells promote functional recovery and neurovascular plasticity after stroke in rats. J Cereb Blood Flow Metab. 2013;33(11):1711–1715.
- Jin Q, Cheng J, Liu Y, et al. Improvement of functional recovery by chronic metformin treatment is associated with enhanced alternative activation of microglia/macrophages and increased angiogenesis and neurogenesis following experimental stroke. Brain Behav Immun. 2014;40:131–142.
- Fan Y, Xiong X, Zhang Y, et al. MKEY, a peptide inhibitor of CXCL4-CCL5 heterodimer formation, protects against stroke in mice. J Am Heart Assoc. 2016;5(9):e003615.
- Nie L, Wang S, Wang X, et al. In vivo volumetric photoacoustic molecular angiography and therapeutic monitoring with targeted plasmonic nanostars. Small. 2014;10(8):1585–1593.
- Nie L, Huang P, Li W, et al. Early-stage imaging of nanocarrier-enhanced chemotherapy response in living subjects by scalable photoacoustic microscopy. ACS Nano. 2014;8(12):12141–12150.
- Qian X, Zhao P, Li W, et al. MicroRNA-26a promotes tumor growth and angiogenesis in glioma by directly targeting prohibitin. CNS Neurosci Ther. 2013;19(10):804–812.
- Li P, Mao L, Zhou G, et al. Adoptive regulatory T-cell therapy preserves systemic immune homeostasis after cerebral ischemia. Stroke. 2013; 44(12):3509–3515.
- Jiang C, Wang J, Yu L, et al. Comparison of the therapeutic effects of bone marrow mononuclear cells and microglia for permanent cerebral ischemia. Behav Brain Res. 2013;250:222–229.
- Wang H, Nagai A, Sheikh AM, et al. Human mesenchymal stem cell transplantation changes proinflammatory gene expression through a nuclear factor-kappaB-dependent pathway in a rat focal cerebral ischemic model. J Neurosci Res. 2013;91(11):1440–1449.
- Patkar S, Tate R, Modo M, Plevin R, Carswell HV. Conditionally immortalized neural stem cells promote functional recovery and brain plasticity after transient focal cerebral ischemia in mice. Stem Cell Res. 2012;8(1):14–25.
- Arai K, Jin G, Navaratna D, Lo EH. Brain angiogenesis in developmental and pathological processes: Neurovascular injury and angiogenic recovery after stroke. FEBS J. 2009;276(17):4644–4652.
- Jetten N, Verbruggen S, Gijbels MJ, Post MJ, De Winther MP, Donners MM. Anti-inflammatory M2, but not pro-inflammatory M1 macrophages promote angiogenesis in vivo. Angiogenesis. 2014;17(1):109–118.
- Chen J, Ning R, Zacharek A, et al. MiR-126 contributes to human umbilical cord blood cell-induced neurorestorative effects after stroke in type-2 diabetic mice. Stem Cells. 2016;34(1):102–113.
- Kang K, Ma R, Cai W, et al. Exosomes secreted from CXCR4 overexpressing mesenchymal stem cells promote cardioprotection via Akt signaling pathway following myocardial infarction. Stem Cells Int. 2015;2015:659890.
- Mineo M, Garfield SH, Taverna S, et al. Exosomes released by K562 chronic myeloid leukemia cells promote angiogenesis in a Src-dependent fashion. Angiogenesis. 2012;15(1):33–45.
- Lazar I, Clement E, Ducoux-Petit M, et al. Proteome characterization of melanoma exosomes reveals a specific signature for metastatic cell lines. Pigment Cell Melanoma Res. 2015;28(4):464–475.
- Gleissner CA, Shaked I, Little KM, Ley K. CXC chemokine ligand 4 induces a unique transcriptome in monocyte-derived macrophages. J Immunol. 2010;184(9):4810–4818.
- Hannafon BN, Carpenter KJ, Berry WL, Janknecht R, Dooley WC, Ding WQ. Exosome-mediated microRNA signaling from breast cancer cells is altered by the anti-angiogenesis agent docosahexaenoic acid (DHA). Mol Cancer. 2015;14:133.
- Ma DN, Chai ZT, Zhu XD, et al. MicroRNA-26a suppresses epithelial-mesenchymal transition in human hepatocellular carcinoma by repressing enhancer of zeste homolog 2. J Hematol Oncol. 2016;9:1.
- Zgheib C, Liechty KW. Shedding light on miR-26a: Another key regulator of angiogenesis in diabetic wound healing. J Mol Cell Cardiol. 2016;92:203–205.
- Garcia NA, Ontoria-Oviedo I, Gonzalez-King H, Diez-Juan A, Sepulveda P. Glucose starvation in cardiomyocytes enhances exosome secretion and promotes angiogenesis in endothelial cells. PLOS ONE. 2015;10(9):e0138849.
- King HW, Michael, MZ, Gleadle JM. Hypoxic enhancement of exosome release by breast cancer cells. BMC Cancer. 2012;12:421.
- Kucharzewska P, Christianson HC, Welch JE, et al. Exosomes reflect the hypoxic status of glioma cells and mediate hypoxia-dependent activation of vascular cells during tumor development. Proc Natl Acad Sci U S A. 2013;110(18):7312–7317.
- Mayo JN, Bearden SE. Driving the hypoxia inducible pathway in human pericytes promotes vascular density in an exosome dependent manner. Microcirculation. 2015;22(8):711–723.
- Taylor DD, Gercel-Taylor C. Exosome platform for diagnosis and monitoring of traumatic brain injury. Philos Trans R Soc Lond B Biol Sci. 2014; 369(1652). doi:10.1098/rstb.2013.0503
- Xia CY, Zhang S, Gao Y, Wang ZZ, Chen NH. Selective modulation of microglia polarization to M2 phenotype for stroke treatment. Int Immunopharmacol. 2015;25(2):377–382.