Abstract
Cerebral infarction is one of the most prevalent cerebrovascular disorders. Microglia and infiltrating macrophages play a key role in regulating the inflammatory response after ischemic stroke. Regulation of microglia/macrophages polarization contributes to the recovery of neurological function in cerebral infarction. In recent decades, human umbilical cord blood mononuclear cells (hUCBMNCs) have been considered a potential therapeutic alternative. However, the mechanism of action is yet unclear. Our study aimed to explore whether hUCBMNCs treatment for cerebral infarction is via regulation of microglia/macrophages polarization. Adult male Sprague–Dawley rats were subjected to middle cerebral artery occlusion (MCAO) and were treated by intravenous routine with or without hUCBMNCs at 24 h following MCAO. We evaluated the therapeutic effects of hUCBMNCs on cerebral infarction by measuring animal behavior and infarct volume, and further explored the possible mechanisms of hUCBMNCs for cerebral infarction by measuring inflammatory factors and microglia/macrophages markers using Elisa and immunofluorescence staining, respectively. We found that administration with hUCBMNCs improved behavioral functions and reduced infarct volume. Rats treated with hUCBMNCs showed a significant reduction in the level of IL-6, and TNF-α and increased the level of IL-4 and IL-10 compared to those treated without hUCBMNCs. Furthermore, hUCBMNCs inhibited M1 polarization and promoted M2 polarization of microglia/macrophages after MCAO. We conclude that hUCBMNCs could ameliorate cerebral brain injury by promoting microglia/macrophages M2 polarization in MCAO Rats. This experiment provides evidence that hUCBMNCs represent a promising therapeutic option for ischemic stroke.
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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
References
Breckwoldt MO, Chen JW, Stangenberg L, Aikawa E, Rodriguez E, Qiu S, Moskowitz MA, Weissleder R (2008) Tracking the inflammatory response in stroke in vivo by sensing the enzyme myeloperoxidase. Proc Natl Acad Sci 105(47):18584–18589. https://doi.org/10.1073/pnas.0803945105
Can A, Karahuseyinoglu S (2007) Concise review: human umbilical cord stroma with regard to the source of fetus-derived stem cells. Stem Cells 25(11):2886–2895. https://doi.org/10.1634/stemcells.2007-0417
Chen J, Sanberg PR, Li Y, Wang L, Lu M, Willing AE, Sanchez-Ramos J, Chopp M (2001) Intravenous administration of human umbilical cord blood reduces behavioral deficits after stroke in rats. Stroke 32(11):2682–2688. https://doi.org/10.1161/hs1101.098367
Chu HX, Broughton BRS, Ah Kim H, Lee S, Drummond GR, Sobey CG (2015) Evidence that Ly6Chi monocytes are protective in acute ischemic stroke by promoting M2 macrophage polarization. Stroke 46(7):1929–1937. https://doi.org/10.1161/STROKEAHA.115.009426
De Meyer SF, Denorme F, Langhauser F, Geuss E, Fluri F, Kleinschnitz C (2016) Thromboinflammation in stroke brain damage. Stroke 47(4):1165–1172. https://doi.org/10.1161/STROKEAHA.115.011238
Dong C, Chen M, Cai B, Zhang C, Xiao G, Luo W (2022) Mesenchymal stem cell-derived exosomes improved cerebral infarction via transferring miR-23a-3p to activate microglia. NeuroMol Med 24(3):290–298. https://doi.org/10.1007/s12017-021-08686-8
Ginhoux F, Greter M, Leboeuf M, Nandi S, See P, Gokhan S, Mehler MF, Conway SJ, Merad M (2010) Fate mapping analysis reveals that adult microglia derive from primitive macrophages. Science 330(6005):841–845. https://doi.org/10.1126/science.1194637
Gritti A, Bonfanti L, Doetsch F, Caille I, Alvarez-Buylla A, Lim DA, Galli R, Verdugo JMG, Vescovi AL (2002) Multipotent neural stem cells reside into the rostral extension and olfactory bulb of adult rodents. J Neurosci 22(2):437–445. https://doi.org/10.1523/JNEUROSCI.22-02-00437.2002
Gu BJ, Kung DK, Chen HI (2021) Cell therapy for stroke. Neurosurgery 88(4):733–745. https://doi.org/10.1093/neuros/nyaa531
Guan Y, Zhu Y, Liu X, Huang H, Wang Z, Liu B, Zhu Y, Wang Q (2014) Effect of human umbilical cord blood mesenchymal stem cell transplantation on neuronal metabolites in ischemic rabbits. BMC Neurosci. https://doi.org/10.1186/1471-2202-15-41
Hu X, Li P, Guo Y, Wang H, Leak RK, Chen S, Gao Y, Chen J (2012) Microglia/macrophage polarization dynamics reveal novel mechanism of injury expansion after focal cerebral ischemia. Stroke 43(11):3063–3070. https://doi.org/10.1161/STROKEAHA.112.659656
Jayaraj RL, Azimullah S, Beiram R, Jalal FY, Rosenberg GA (2019) Neuroinflammation: friend and foe for ischemic stroke. J Neuroinflamm. https://doi.org/10.1186/s12974-019-1516-2
Jiang L, Womble T, Saporta S, Chen N, Sanberg CD, Sanberg PR, Willing AE (2010) Human umbilical cord blood cells decrease microglial survival in vitro. Stem Cells Dev 19(2):221–228. https://doi.org/10.1089/scd.2009.0170
Jiang CT, Wu WF, Deng YH, Ge JW (2020) Modulators of microglia activation and polarization in ischemic stroke (review). Mol Med Rep. https://doi.org/10.3892/mmr.2020.11003
Jurcau A, Simion A (2022) Neuroinflammation in cerebral ischemia and ischemia/reperfusion injuries: from pathophysiology to therapeutic strategies. Int J Mol Sci 23(1):14. https://doi.org/10.3390/ijms23010014
Karlupia N, Manley NC, Prasad K, Schäfer R, Steinberg GK (2014) Intraarterial transplantation of human umbilical cord blood mononuclear cells is more efficacious and safer compared with umbilical cord mesenchymal stromal cells in a rodent stroke model. Stem Cell Res Therapy. https://doi.org/10.1186/scrt434
Kim G, Subash M, Yoon JS, Jo D, Han J, Hong JM, Kim S, Suh-Kim H (2020) Neurogenin-1 overexpression increases the therapeutic effects of mesenchymal stem cells through enhanced engraftment in an ischemic rat brain. Int J Stem Cells 13(1):127–141. https://doi.org/10.15283/ijsc19111
Kreutzberg GW (1996) Microglia: a sensor for pathological events in the CNS. Trends Neurosci 19(8):312–318. https://doi.org/10.1016/0166-2236(96)10049-7
Lan X, Han X, Li Q, Yang Q, Wang J (2017) Modulators of microglial activation and polarization after intracerebral haemorrhage. Nat Rev Neurol 13(7):420–433. https://doi.org/10.1038/nrneurol.2017.69
Lan Y, Xie H, Jin Q, Zhao X, Shi Y, Zhou Y, Hu Z, Ye Y, Xie Z (2022) Extracellular vesicles derived from neural EGFL-Like 1-modified mesenchymal stem cells improve acellular bone regeneration via the miR-25–5p-SMAD2 signaling axis. Bioact Mater 17:457–470. https://doi.org/10.1016/j.bioactmat.2022.01.019
Liu L, Cen J, Man Y, Li J, Zhang D, Wang F, Li J, Ma J, Ji B (2018) Transplantation of human umbilical cord blood mononuclear cells attenuated ischemic injury in MCAO rats via inhibition of NF-κB and NLRP3 inflammasome. Neuroscience 369:314–324. https://doi.org/10.1016/j.neuroscience.2017.11.027
Liu X, Zhang M, Liu H, Zhu R, He H, Zhou Y, Zhang Y, Li C, Huang G (2021) Bone marrow mesenchymal stem cell-derived exosomes attenuate cerebral ischemia-reperfusion injury-induced neuroinflammation and pyroptosis by modulating microglia M1/M2 phenotypes. Exp Neurol 341:113700. https://doi.org/10.1016/j.expneurol.2021.113700
Longa EZ, Weinstein PR, Carlson S, Cummins R (1989) Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 20(1):84–91. https://doi.org/10.1161/01.str.20.1.84
Ma Y, Wang J, Wang Y, Yang G (2017) The biphasic function of microglia in ischemic stroke. Prog Neurobiol 157:247–272. https://doi.org/10.1016/j.pneurobio.2016.01.005
Mendelson SJ, Prabhakaran S (2021) Diagnosis and management of transient ischemic attack and acute ischemic stroke. JAMA 325(11):1088. https://doi.org/10.1001/jama.2020.26867
Menon LG, Picinich S, Koneru R, Gao H, Lin SY, Koneru M, Mayer-Kuckuk P, Glod J, Banerjee D (2007) Differential gene expression associated with migration of mesenchymal stem cells to conditioned medium from tumor cells or bone marrow cells. Stem Cells 25(2):520–528. https://doi.org/10.1634/stemcells.2006-0257
Nalamolu KR, Venkatesh I, Mohandass A, Klopfenstein JD, Pinson DM, Wang DZ, Kunamneni A, Veeravalli KK (2019) Exosomes secreted by the cocultures of normal and oxygen–glucose-deprived stem cells improve post-stroke outcome. NeuroMol Med 21(4):529–539. https://doi.org/10.1007/s12017-019-08540-y
Pan Y, Nastav JB, Zhang H, Bretton RH, Panneton WM, Bicknese AR (2005) Engraftment of freshly isolated or cultured human umbilical cord blood cells and the effect of cyclosporin A on the outcome. Exp Neurol 192(2):365–372. https://doi.org/10.1016/j.expneurol.2004.11.011
Park HW, Moon H, Kim HR, Paek SL, Kim Y, Chang JW, Yang YS, Kim K, Paek SH (2015) Human umbilical cord blood-derived mesenchymal stem cells improve functional recovery through thrombospondin1, pantraxin3, and vascular endothelial growth factor in the ischemic rat brain. J Neurosci Res 93(12):1814–1825. https://doi.org/10.1002/jnr.23616
Perego C, Fumagalli S, De Simoni M (2011) Temporal pattern of expression and colocalization of microglia/macrophage phenotype markers following brain ischemic injury in mice. J Neuroinflamm. https://doi.org/10.1186/1742-2094-8-174
Pluta R, Januszewski S, Czuczwar SJ (2021) Neuroinflammation in post-ischemic neurodegeneration of the brain: friend, foe, or both? Int J Mol Sci 22(9):4405. https://doi.org/10.3390/ijms22094405
Pourmohammadi-Bejarpasi Z, Roushandeh AM, Saberi A, Rostami MK, Toosi SMR, Jahanian-Najafabadi A, Tomita K, Kuwahara Y, Roudkenar MH (2020) Mesenchymal stem cells-derived mitochondria transplantation mitigates I/R-induced injury, abolishes I/R-induced apoptosis, and restores motor function in acute ischemia stroke rat model. Brain Res Bull 165:70–80. https://doi.org/10.1016/j.brainresbull.2020.09.018
Qin C, Fan W, Liu Q, Shang K, Murugan M, Wu L, Wang W, Tian D (2017) Fingolimod protects against ischemic white matter damage by modulating microglia toward M2 polarization via STAT3 pathway. Stroke 48(12):3336–3346. https://doi.org/10.1161/STROKEAHA.117.018505
Rawlinson C, Jenkins S, Thei L, Dallas ML, Chen R (2020) Post-ischaemic immunological response in the brain: targeting microglia in ischaemic stroke therapy. Brain Sci 10(3):159. https://doi.org/10.3390/brainsci10030159
Ringdén O, Okas M, Uhlin M, Uzunel M, Remberger M, Mattsson J (2008) Unrelated cord blood and mismatched unrelated volunteer donor transplants, two alternatives in patients who lack an HLA-identical donor. Bone Marrow Transplant 42(10):643–648. https://doi.org/10.1038/bmt.2008.239
Rocha V, Labopin M, Sanz G, Arcese W, Schwerdtfeger R, Bosi A, Jacobsen N, Ruutu T, Gluckman E (2004) Transplants of umbilical-cord blood or bone marrow from unrelated donors in adults with acute leukemia. N Engl J Med 351(22):2276–2285. https://doi.org/10.1056/NEJMoa041469
Rogers DC, Campbell CA, Stretton JL, Mackay KB (1997) Correlation between motor impairment and infarct volume after permanent and transient middle cerebral artery occlusion in the rat. Stroke 28(10):2060–2066. https://doi.org/10.1161/01.str.28.10.2060. (discussion 2066)
Sahota P, Vahidy F, Nguyen C, Bui T, Yang B, Parsha K, Garrett J, Bambhroliya A, Savitz S (2013) Changes in spleen size in patients with acute ischemic stroke: a pilot observational study. Int J Stroke 8(2):60–67. https://doi.org/10.1111/ijs.12022
Saino O, Taguchi A, Nakagomi T, Nakano-Doi A, Kashiwamura S, Doe N, Nakagomi N, Soma T, Matsuyama T (2010) Immunodeficiency reduces neural stem/progenitor cell apoptosis and enhances neurogenesis in the cerebral cortex after stroke. J Neurosci Res. https://doi.org/10.1002/jnr.22410
Shiao ML, Yuan C, Crane AT, Voth JP, Juliano M, Stone LLH, Nan Z, Zhang Y, Low WC (2019) Immunomodulation with human umbilical cord blood stem cells ameliorates ischemic brain injury – a brain transcriptome profiling analysis. Cell Transplant 28(7):864–873. https://doi.org/10.1177/0963689719836763
Song S, Luo L, Sun B, Sun D (2019) Roles of glial ion transporters in brain diseases. Glia 68(3):472–494. https://doi.org/10.1002/glia.23699
Stone LLH, Xiao F, Rotschafer J, Nan Z, Juliano M, Sanberg CD, Sanberg PR, Kuzmin-Nichols N, Low WC (2016) Amelioration of ischemic brain injury in rats with human umbilical cord blood stem cells: mechanisms of action. Cell Transplant 25(8):1473–1488. https://doi.org/10.3727/096368916X691277
Suda S, Nito C, Yokobori S, Sakamoto Y, Nakajima M, Sowa K, Obinata H, Sasaki K, Kimura K (2020) Recent advances in cell-based therapies for ischemic stroke. Int J Mol Sci 21(18):6718. https://doi.org/10.3390/ijms21186718
Tucker LB, Velosky AG, McCabe JT (2018) Applications of the Morris water maze in translational traumatic brain injury research. Neurosci Biobehav Rev 88:187–200. https://doi.org/10.1016/j.neubiorev.2018.03.010
Vendrame M, Cassady J, Newcomb J, Butler T, Pennypacker KR, Zigova T, Davis Sanberg C, Sanberg PR, Willing AE (2004) Infusion of human umbilical cord blood cells in a rat model of stroke dose-dependently rescues behavioral deficits and reduces infarct volume. Stroke 35(10):2390–2395. https://doi.org/10.1161/01.STR.0000141681.06735.9b
Venkat P, Shen Y, Chopp M, Chen J (2018) Cell-based and pharmacological neurorestorative therapies for ischemic stroke. Neuropharmacology 134:310–322. https://doi.org/10.1016/j.neuropharm.2017.08.036
Virani SS, Alonso A, Benjamin EJ, Bittencourt MS, Callaway CW, Carson AP, Chamberlain AM, Chang AR, Tsao CW (2020) Heart disease and stroke statistics—2020 update: a report from the American Heart Association. Circulation. https://doi.org/10.1161/CIR.0000000000000757
Wang Z, He D, Zeng Y, Zhu L, Yang C, Lu Y, Huang J, Cheng X, Tan X (2019) The spleen may be an important target of stem cell therapy for stroke. J Neuroinflamm. https://doi.org/10.1186/s12974-019-1400-0
Womble TA, Green S, Shahaduzzaman M, Grieco J, Sanberg PR, Pennypacker KR, Willing AE (2014) Monocytes are essential for the neuroprotective effect of human cord blood cells following middle cerebral artery occlusion in rat. Mol Cell Neurosci 59:76–84. https://doi.org/10.1016/j.mcn.2014.01.004
Wu W, Chen X, Hu C, Li J, Yu Z, Cai W (2010) Transplantation of neural stem cells expressing hypoxia-inducible factor-1α (HIF-1α) improves behavioral recovery in a rat stroke model. J Clin Neurosci 17(1):92–95. https://doi.org/10.1016/j.jocn.2009.03.039
Xiong X, Liu L, Yang Q (2016) Functions and mechanisms of microglia/macrophages in neuroinflammation and neurogenesis after stroke. Prog Neurobiol 142:23–44. https://doi.org/10.1016/j.pneurobio.2016.05.001
Xu S, Lu J, Shao A, Zhang JH, Zhang J (2020) Glial Cells: role of the immune response in ischemic stroke. Front Immunol 11:294. https://doi.org/10.3389/fimmu.2020.00294
Yang J, Wang Z, Liu X, Lu P (2021) Modulation of vascular integrity and neuroinflammation by peroxiredoxin 4 following cerebral ischemia-reperfusion injury. Microvasc Res 135:104144. https://doi.org/10.1016/j.mvr.2021.104144
Ye Y, Chang Z, Wang P, Wang Y, Liang J, Chen C, Wang J, Sun H, Li X (2022) Infarct-preconditioning exosomes of umbilical cord mesenchymal stem cells promoted vascular remodeling and neurological recovery after stroke in rats. Stem Cell Res Therapy. https://doi.org/10.1186/s13287-022-03083-9
Zhai L, Maimaitiming Z, Cao X, Xu Y, Jin J (2019) Nitrogen-doped carbon nanocages and human umbilical cord mesenchymal stem cells cooperatively inhibit neuroinflammation and protect against ischemic stroke. Neurosci Lett 708:134346. https://doi.org/10.1016/j.neulet.2019.134346
Zhang BJ, Men XJ, Lu ZQ, Li HY, Qiu W, Hu XQ (2013) Splenectomy protects experimental rats from cerebral damage after stroke due to anti-inflammatory effects. Chin Med J (engl) 126(12):2354–2360
Zhang B, Zhang H, Shi S, Bai Y, Zhe X, Zhang S, Li Y (2019) Interleukin-11 treatment protected against cerebral ischemia/reperfusion injury. Biomed Pharmacother 115:108816. https://doi.org/10.1016/j.biopha.2019.108816
Zhang Z, Zou X, Zhang R, Xie Y, Feng Z, Li F, Han J, Sun H, Jiang X (2021) Human umbilical cord mesenchymal stem cell-derived exosomal miR-146a-5p reduces microglial-mediated neuroinflammation via suppression of the IRAK1/TRAF6 signaling pathway after ischemic stroke. Aging 13(2):3060–3079. https://doi.org/10.18632/aging.202466
Zhao Q, Hu J, Xiang J, Gu Y, Jin P, Hua F, Zhang Z, Liu Y, Ye X (2015) Intranasal administration of human umbilical cord mesenchymal stem cells-conditioned medium enhances vascular remodeling after stroke. Brain Res 1624:489–496. https://doi.org/10.1016/j.brainres.2015.08.003
Zhu L, He D, Han L, Cao H (2015) Stroke research in china over the past decade: analysis of NSFC funding. Transl Stroke Res 6(4):253–256. https://doi.org/10.1007/s12975-015-0404-z
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This study would like to thank Shandong Cord Blood Bank for providing human umbilical cord blood mononuclear cells.
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Conceptualization: SG; methodology: SG; material preparation: GL, GX, and XT. data collection and analysis: HL, GX, and XT; writing—original draft preparation: HL; writing—review and editing: SG, HL, and YX; supervision: SG. All authors commented on previous versions of the manuscript and approved the final manuscript.
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Li, H., Xiao, G., Tan, X. et al. Human umbilical cord blood mononuclear cells ameliorate ischemic brain injury via promoting microglia/macrophages M2 polarization in MCAO Rats. Exp Brain Res 241, 1585–1598 (2023). https://doi.org/10.1007/s00221-023-06600-1
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DOI: https://doi.org/10.1007/s00221-023-06600-1