Abstract
To investigate the association of miR-106b-5p with neuroinflammation and microglial activation in a status epilepticus (SE) mouse model. We examined changes in the expression of microRNA-106b-5p (miRNA-106b-5p), repulsive guidance molecule A (RGMa), triggering receptor expressed on myeloid cells 2 (TREM2), and the microglia-related markers interleukin (IL)-1β, IL-4, IL-6, IL-10, inducible nitric oxide synthase (iNOS), and arginase-1 (Arg-1) in the mouse hippocampus of the lithium–pilocarpine-induced SE mouse model. Eighty-four female C57BL/6 mice were randomly divided into a normal control group (n = 12), and six SE groups (n = 12/group), which were monitored at 6 h and at 1, 3, 7, 14, and 21 days (d) post-SE induction. Unlike in the dentate gyrus, immunohistochemical staining revealed prominent neuronal swelling at 6 h, significant neuronal loss and apoptosis on day 3, and recovery by day 14 in the hippocampal cornu ammonis (CA)1 and CA3 pyramidal cells in SE mice. We noted elevated levels of miRNA-106b-5p and all microglia-related markers, which peaked at 3 days post-SE, except IL-4, which peaked at 7 days post-SE, indicating inflammation and microglial activation. RGMa and TREM2 levels decreased at 6 h post-SE. All markers but miRNA-106b-5p, RGMa, and TREM2 returned to baseline levels at 21 days post-SE. Dual luciferase reporter gene assay showed that microRNA-106b-5p can interact with RGMa. We observed that miR-106b-5p level increased while both RGMa and TREM2 levels decreased post-SE and showed associations with microglial activation and inflammation in the mouse hippocampus, suggesting their potential as SE therapeutic targets.
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The datasets generated and analyzed during the current study are available from the corresponding author on reasonable request.
References
An N, Zhao W, Liu Y, Yang X, Chen P (2016) Elevated serum miR-106b and miR-146a in patients with focal and generalized epilepsy. Epilepsy Res 127:311–316. https://doi.org/10.1016/j.eplepsyres.2016.09.019
Avignone E, Ulmann L, Levavasseur F, Rassendren F, Audinat E (2008) Status epilepticus induces a particular microglial activation state characterized by enhanced purinergic signaling. J Neurosci 28:9133–9144. https://doi.org/10.1523/jneurosci.1820-08.2008
Benson MJ, Manzanero S, Borges K (2015) Complex alterations in microglial M1/M2 markers during the development of epilepsy in two mouse models. Epilepsia 56:895–905. https://doi.org/10.1111/epi.12960
Brennan GP, Henshall DC (2020) MicroRNAs as regulators of brain function and targets for treatment of epilepsy. Nat Rev Neurol 16:506–519. https://doi.org/10.1038/s41582-020-0369-8
Brindley E, Hill TDM, Henshall DC (2019) MicroRNAs as biomarkers and treatment targets in status epilepticus. Epilepsy Behav 101:106272. https://doi.org/10.1016/j.yebeh.2019.04.025
Da Mesquita S, Kipnis J (2017) DAMed in (Trem) 2 steps. Cell 169:1172–1174. https://doi.org/10.1016/j.cell.2017.05.039
Deming Y, Li Z, Benitez BA, Cruchaga C (2018) Triggering receptor expressed on myeloid cells 2 (TREM2): a potential therapeutic target for Alzheimer disease? Expert Opin Ther Targets 22:587–598. https://doi.org/10.1080/14728222.2018.1486823
Fiest KM, Sauro KM, Wiebe S, Patten SB, Kwon CS, Dykeman J, Pringsheim T, Lorenzetti DL, Jetté N (2017) Prevalence and incidence of epilepsy: a systematic review and meta-analysis of international studies. Neurology 88:296–303. https://doi.org/10.1212/wnl.0000000000003509
Gisslén M, Heslegrave A, Veleva E, Yilmaz A, Andersson LM, Hagberg L, Spudich S, Fuchs D, Price RW, Zetterberg H (2019) CSF concentrations of soluble TREM2 as a marker of microglial activation in HIV-1 infection. Neurol Neuroimmunol Neuroinflamm 6:e512. https://doi.org/10.1212/nxi.0000000000000512
Gupta J, Bromwich M, Radell J, Arshad MN, Gonzalez S, Luikart BW, Aaron GB, Naegele JR (2019) Restrained dendritic growth of adult-born granule cells innervated by transplanted fetal GABAergic interneurons in mice with temporal lobe epilepsy. eNeuro. https://doi.org/10.1523/eneuro.0110-18.2019
Hiragi T, Ikegaya Y, Koyama R (2018) Microglia after seizures and in epilepsy. Cells. https://doi.org/10.3390/cells7040026
Jay TR, von Saucken VE, Landreth GE (2017) TREM2 in neurodegenerative diseases. Mol Neurodegener 12:56. https://doi.org/10.1186/s13024-017-0197-5
Jiang Y, Li Z, Ma H, Cao X, Liu F, Tian A, Sun X, Li X, Wang J (2018) Upregulation of TREM2 ameliorates neuroinflammatory responses and improves cognitive deficits triggered by surgical trauma in Appswe/PS1dE9 Mice. Cell Physiol Biochem 46:1398–1411. https://doi.org/10.1159/000489155
Jin X, Liu MY, Zhang DF, Zhong X, Du K, Qian P, Gao H, Wei MJ (2019) Natural products as a potential modulator of microglial polarization in neurodegenerative diseases. Pharmacol Res 145:104253. https://doi.org/10.1016/j.phrs.2019.104253
Kitayama M, Ueno M, Itakura T, Yamashita T (2011) Activated microglia inhibit axonal growth through RGMa. PLoS ONE 6:e25234. https://doi.org/10.1371/journal.pone.0025234
Li P, Shen M, Gao F, Wu J, Zhang J, Teng F, Zhang C (2017) An antagomir to microRNA-106b-5p ameliorates cerebral ischemia and reperfusion injury in rats via inhibiting apoptosis and oxidative stress. Mol Neurobiol 54:2901–2921. https://doi.org/10.1007/s12035-016-9842-1
Liu JT, Wu SX, Zhang H, Kuang F (2018) Inhibition of MyD88 signaling skews microglia/macrophage polarization and attenuates neuronal apoptosis in the hippocampus after status epilepticus in mice. Neurotherapeutics 15:1093–1111. https://doi.org/10.1007/s13311-018-0653-0
Liu AH, Chu M, Wang YP (2019) Up-regulation of Trem2 inhibits hippocampal neuronal apoptosis and alleviates oxidative stress in epilepsy via the PI3K/Akt pathway in mice. Neurosci Bull 35:471–485. https://doi.org/10.1007/s12264-018-0324-5
Liu W, Taso O, Wang R et al (2020) Trem2 promotes anti-inflammatory responses in microglia and is suppressed under pro-inflammatory conditions. Hum Mol Genet 29:3224–3248. https://doi.org/10.1093/hmg/ddaa209
Luo C, Koyama R, Ikegaya Y (2016) Microglia engulf viable newborn cells in the epileptic dentate gyrus. Glia 64:1508–1517. https://doi.org/10.1002/glia.23018
Morris G, Reschke CR, Henshall DC (2019) Targeting microRNA-134 for seizure control and disease modification in epilepsy. EBioMedicine 45:646–654. https://doi.org/10.1016/j.ebiom.2019.07.008
Mosser CA, Baptista S, Arnoux I, Audinat E (2017) Microglia in CNS development: shaping the brain for the future. Prog Neurobiol 149–150:1–20. https://doi.org/10.1016/j.pneurobio.2017.01.002
Müller CJ, Bankstahl M, Gröticke I, Löscher W (2009) Pilocarpine vs. lithium-pilocarpine for induction of status epilepticus in mice: development of spontaneous seizures, behavioral alterations and neuronal damage. Eur J Pharmacol 619:15–24. https://doi.org/10.1016/j.ejphar.2009.07.020
Muramatsu R, Kubo T, Mori M et al (2011) RGMa modulates T cell responses and is involved in autoimmune encephalomyelitis. Nat Med 17:488–494. https://doi.org/10.1038/nm.2321
Nakatomi H, Kuriu T, Okabe S, Yamamoto S, Hatano O, Kawahara N, Tamura A, Kirino T, Nakafuku M (2002) Regeneration of hippocampal pyramidal neurons after ischemic brain injury by recruitment of endogenous neural progenitors. Cell 110:429–441. https://doi.org/10.1016/s0092-8674(02)00862-0
Racine RJ (1972) Modification of seizure activity by electrical stimulation. II. Motor seizure. Electroencephalogr Clin Neurophysiol 32:281–294. https://doi.org/10.1016/0013-4694(72)90177-0
Reddy DS, Kuruba R (2013) Experimental models of status epilepticus and neuronal injury for evaluation of therapeutic interventions. Int J Mol Sci 14:18284–18318. https://doi.org/10.3390/ijms140918284
Schafer DP, Lehrman EK, Stevens B (2013) The “quad-partite” synapse: microglia-synapse interactions in the developing and mature CNS. Glia 61:24–36. https://doi.org/10.1002/glia.22389
Shibley H, Smith BN (2002) Pilocarpine-induced status epilepticus results in mossy fiber sprouting and spontaneous seizures in C57BL/6 and CD-1 mice. Epilepsy Res 49:109–120. https://doi.org/10.1016/s0920-1211(02)00012-8
Siebold C, Yamashita T, Monnier PP, Mueller BK, Pasterkamp RJ (2017) RGMs: structural insights, molecular regulation, and downstream signaling. Trends Cell Biol 27:365–378. https://doi.org/10.1016/j.tcb.2016.11.009
Sirven JI (2015) Epilepsy: a spectrum disorder. Cold Spring Harb Perspect Med 5:a022848. https://doi.org/10.1101/cshperspect.a022848
Song M, Tian F, Xia H, Xie Y (2019) Repulsive guidance molecule a suppresses seizures and mossy fiber sprouting via the FAK-p120RasGAP-Ras signaling pathway. Mol Med Rep 19:3255–3262. https://doi.org/10.3892/mmr.2019.9951
Sticht C, De La Torre C, Parveen A, Gretz N (2018) miRWalk: an online resource for prediction of microRNA binding sites. PLoS ONE 13:e0206239. https://doi.org/10.1371/journal.pone.0206239
Therajaran P, Hamilton JA, O’Brien TJ, Jones NC, Ali I (2020) Microglial polarization in posttraumatic epilepsy: potential mechanism and treatment opportunity. Epilepsia 61:203–215. https://doi.org/10.1111/epi.16424
Trinka E, Cock H, Hesdorffer D, Rossetti AO, Scheffer IE, Shinnar S, Shorvon S, Lowenstein DH (2015) A definition and classification of status epilepticus—report of the ILAE task force on classification of status epilepticus. Epilepsia 56:1515–1523. https://doi.org/10.1111/epi.13121
Ulrich JD, Ulland TK, Colonna M, Holtzman DM (2017) Elucidating the role of TREM2 in Alzheimer’s disease. Neuron 94:237–248. https://doi.org/10.1016/j.neuron.2017.02.042
Upadhya D, Kodali M, Gitai D et al (2019) A model of chronic temporal lobe epilepsy presenting constantly rhythmic and robust spontaneous seizures, co-morbidities and hippocampal neuropathology. Aging Dis 10:915–936. https://doi.org/10.14336/ad.2019.0720
Verrotti A, Iapadre G, Di Francesco L, Zagaroli L, Farello G (2020) Diet in the treatment of epilepsy: what we know so far. Nutrients. https://doi.org/10.3390/nu12092645
Vezzani A, French J, Bartfai T, Baram TZ (2011) The role of inflammation in epilepsy. Nat Rev Neurol 7:31–40. https://doi.org/10.1038/nrneurol.2010.178
Voet S, Prinz M, van Loo G (2019) Microglia in central nervous system inflammation and multiple sclerosis pathology. Trends Mol Med 25:112–123. https://doi.org/10.1016/j.molmed.2018.11.005
Walker DG, Lue LF (2015) Immune phenotypes of microglia in human neurodegenerative disease: challenges to detecting microglial polarization in human brains. Alzheimers Res Ther 7:56. https://doi.org/10.1186/s13195-015-0139-9
Wang J, Zhao J (2021) MicroRNA dysregulation in epilepsy: from pathogenetic involvement to diagnostic biomarker and therapeutic agent development. Front Mol Neurosci 14:650372. https://doi.org/10.3389/fnmol.2021.650372
Wang J, Yu JT, Tan L et al (2015) Genome-wide circulating microRNA expression profiling indicates biomarkers for epilepsy. Sci Rep 5:9522. https://doi.org/10.1038/srep09522
Xu X, Gao Y, Shan F, Feng J (2016) A novel role for RGMa in modulation of bone marrow-derived dendritic cells maturation induced by lipopolysaccharide. Int Immunopharmacol 33:99–107. https://doi.org/10.1016/j.intimp.2016.02.008
Yu T, Yu H, Zhang B, Wang D, Li B, Zhu J, Zhu W (2019) Promising neuroprotective function for M2 microglia in kainic acid-induced neurotoxicity via the down-regulation of NF-κB and caspase 3 signaling pathways. Neuroscience 406:86–96. https://doi.org/10.1016/j.neuroscience.2019.03.002
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This work was supported by the National Key Research and Development Plan (Grant No. 2016YFC1306203).
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HW and TY conceived and designed research; TY, HF and DRD collected data and conducted research; TY and JJS analyzed and interpreted data; TY wrote the initial paper; HW revised the paper; HW had primary responsibility for the final content. All authors read and approved the final manuscript.
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All animal experiments were performed according to the National Institutes of Health guidelines for the Care and Use of Laboratory Animals (NIH publication 80-23, revised in 1996) and the necessary approval (2020PS505K) was obtained from the Animal Ethics Committee of Shengjing Hospital Affiliated with China Medical University.
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Yu, T., Fu, H., Sun, JJ. et al. miR-106b-5p upregulation is associated with microglial activation and inflammation in the mouse hippocampus following status epilepticus. Exp Brain Res 239, 3315–3325 (2021). https://doi.org/10.1007/s00221-021-06208-3
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DOI: https://doi.org/10.1007/s00221-021-06208-3