Generic placeholder image

Current Neurovascular Research

Author(s): Renbo Li, Jian Han, Bo Chen and Jingbo Shang*

DOI: 10.2174/1567202619666220601111715

DownloadDownload PDF Flyer Cite As
Homeodomain Interacting Protein Kinase 2-Modified Rat Spinal Astrocytes Affect Neurofunctional Recovery After Spinal Cord Injury

Page: [171 - 180] Pages: 10

  • * (Excluding Mailing and Handling)

Abstract

Background: Spinal cord injury (SCI) is regarded as an acute neurological disorder, and astrocytes play a role in the progression of SCI.

Objective: Herein, we investigated the roles of homeodomain-interacting protein kinase 2 (HIPK2)- modified rat spinal astrocytes in neurofunctional recovery after SCI.

Methods: Rat spinal astrocytes were cultured, isolated, and then identified through microscopic observation and immunofluorescence staining. Astrocytes were infected with the adenovirus vector overexpressing HIPK2 for modification, and proliferation and apoptosis of astrocytes were examined using Cell Counting Kit-8 method and flow cytometry. SCI rat models were established and treated with astrocytes or HIPK2-modified astrocytes. Subsequently, rat motor ability was analyzed via the Basso-Beattie-Bresnahan (BBB) scoring and inclined-plane test, and the damage to spinal cord tissues and neuronal survival were observed via Hematoxylin-eosin staining and Nissl staining. The levels of HIPK2, brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and nuclear factor erythroid 2- related transcription factor 2 (Nrf2)/antioxidant response element (ARE) pathway-related proteins were detected.

Results: Rat spinal astrocytes were harvested successfully. HIPK2 overexpression accelerated the proliferation and repressed the apoptosis of rat spinal astrocytes. Rat spinal astrocytes treatment increased BBB points and the maximum angle at which SCI rats remained stable, ameliorated damage to spinal cord tissues, increased the number of neurons, and attenuated neural damage and inflammation, while the treatment of HIPK2-modified rat spinal astrocytes imparted more pronounced effects to the neurofunctional recovery of SCI rats. Meanwhile, HIPK2-modified rat spinal astrocytes further activated the Nrf2/ARE pathway.

Conclusion: HIPK2-modified rat spinal astrocytes facilitated neurofunctional recovery and activated the Nrf2/ARE pathway after SCI.

Keywords: Spinal cord injury, astrocytes, HIPK2, neurofunctional recovery, Nrf2/ARE pathway, gene modification.

[1]
Rodríguez-Barrera R, Rivas-González M, García-Sánchez J, Mojica-Torres D, Ibarra A. Neurogenesis after spinal cord injury: State of the art. Cells 2021; 10(6): 1499.
[http://dx.doi.org/10.3390/cells10061499] [PMID: 34203611]
[2]
Sun F, Li SG, Zhang HW, Hua FW, Sun GZ, Huang Z. MiRNA-411 attenuates inflammatory damage and apoptosis following spinal cord injury. Eur Rev Med Pharmacol Sci 2020; 24(2): 491-8.
[PMID: 32016950]
[3]
Hachem LD, Fehlings MG. Pathophysiology of spinal cord injury. Neurosurg Clin N Am 2021; 32(3): 305-13.
[http://dx.doi.org/10.1016/j.nec.2021.03.002] [PMID: 34053718]
[4]
Eli I, Lerner DP, Ghogawala Z. Acute traumatic spinal cord injury. Neurol Clin 2021; 39(2): 471-88.
[http://dx.doi.org/10.1016/j.ncl.2021.02.004] [PMID: 33896529]
[5]
Zimmermann R, Vieira Alves Y, Sperling LE, Pranke P. Nanotechnology for the treatment of spinal cord injury. Tissue Eng Part B Rev 2021; 27(4): 353-65.
[http://dx.doi.org/10.1089/ten.teb.2020.0188] [PMID: 33135599]
[6]
Zhou CL, Li F, Wu XW, et al. Overexpression of miRNA-433-5p protects acute spinal cord injury through activating MAPK1. Eur Rev Med Pharmacol Sci 2020; 24(6): 2829-35.
[PMID: 32271400]
[7]
Sancho L, Contreras M, Allen NJ. Glia as sculptors of synaptic plasticity. Neurosci Res 2021; 167: 17-29.
[http://dx.doi.org/10.1016/j.neures.2020.11.005] [PMID: 33316304]
[8]
Molofsky AV, Deneen B. Astrocyte development: A guide for the perplexed. Glia 2015; 63(8): 1320-9.
[http://dx.doi.org/10.1002/glia.22836] [PMID: 25963996]
[9]
Lukacova N, Kisucka A, Kiss Bimbova K, et al. Glial-neuronal interactions in pathogenesis and treatment of spinal cord injury. Int J Mol Sci 2021; 22(24): 13577.
[http://dx.doi.org/10.3390/ijms222413577] [PMID: 34948371]
[10]
Huang X, Gu YK, Cheng XY, Su ZD. Astrocytes as therapeutic targets after spinal cord injury. Sheng Li Xue Bao 2017; 69(6): 794-804.
[PMID: 29270596]
[11]
Li L, Acioglu C, Heary RF, Elkabes S. Role of astroglial toll-like receptors (TLRs) in central nervous system infections, injury and neurodegenerative diseases. Brain Behav Immun 2021; 91: 740-55.
[http://dx.doi.org/10.1016/j.bbi.2020.10.007] [PMID: 33039660]
[12]
Hashimoto K, Tsuji Y. Arsenic-induced activation of the homeodomain-interacting protein kinase 2 (HIPK2) to cAMP-response element binding protein (CREB) axis. J Mol Biol 2017; 429(1): 64-78.
[http://dx.doi.org/10.1016/j.jmb.2016.11.015] [PMID: 27884605]
[13]
Shang Y, Zhang J, Huang EJ. HIPK2-mediated transcriptional control of NMDA receptor subunit expression regulates neuronal survival and cell death. J Neurosci 2018; 38(16): 4006-19.
[http://dx.doi.org/10.1523/JNEUROSCI.3577-17.2018] [PMID: 29581378]
[14]
Li R, Shang J, Zhou W, Jiang L, Xie D, Tu G. Overexpression of HIPK2 attenuates spinal cord injury in rats by modulating apoptosis, oxidative stress, and inflammation. Biomed Pharmacother 2018; 103: 127-34.
[http://dx.doi.org/10.1016/j.biopha.2018.03.117] [PMID: 29649627]
[15]
Vasconcelos AR, Dos Santos NB, Scavone C, Munhoz CD. Nrf2/ARE pathway modulation by dietary energy regulation in neurological disorders. Front Pharmacol 2019; 10: 33.
[http://dx.doi.org/10.3389/fphar.2019.00033] [PMID: 30778297]
[16]
Chu SF, Zhang Z, Zhou X, et al. Ginsenoside Rg1 protects against ischemic/reperfusion-induced neuronal injury through miR-144/Nrf2/ARE pathway. Acta Pharmacol Sin 2019; 40(1): 13-25.
[http://dx.doi.org/10.1038/s41401-018-0154-z] [PMID: 30262824]
[17]
Wang X, de Rivero Vaccari JP, Wang H, et al. Activation of the nuclear factor E2-related factor 2/antioxidant response element pathway is neuroprotective after spinal cord injury. J Neurotrauma 2012; 29(5): 936-45.
[http://dx.doi.org/10.1089/neu.2011.1922] [PMID: 21806470]
[18]
Dang X, Zhang R, Peng Z, et al. HIPK2 overexpression relieves hypoxia/reoxygenation-induced apoptosis and oxidative damage of cardiomyocytes through enhancement of the Nrf2/ARE signaling pathway. Chem Biol Interact 2020; 316: 108922.
[http://dx.doi.org/10.1016/j.cbi.2019.108922] [PMID: 31837296]
[19]
Li J, Lian ZG, Xu YH, et al. Downregulation of nuclear protein-1 induces cell cycle arrest in G0/G1 phase in glioma cells in vivo and in vitro via P27. Neoplasma 2020; 67(4): 843-50.
[http://dx.doi.org/10.4149/neo_2020_190814N759] [PMID: 32266819]
[20]
Cong Y, Wang C, Wang J, Li H, Li Q. NT-3 promotes oligodendrocyte proliferation and nerve function recovery after spinal cord injury by inhibiting autophagy pathway. J Surg Res 2020; 247: 128-35.
[http://dx.doi.org/10.1016/j.jss.2019.10.033] [PMID: 31776022]
[21]
Lu T, Peng W, Liang Y, et al. PTEN-silencing combined with ChABC-overexpression in adipose-derived stem cells promotes functional recovery of spinal cord injury in rats. Biochem Biophys Res Commun 2020; 532(3): 420-6.
[http://dx.doi.org/10.1016/j.bbrc.2020.08.085] [PMID: 32888649]
[22]
Duan HQ, Wu QL, Yao X, et al. Nafamostat mesilate attenuates inflammation and apoptosis and promotes locomotor recovery after spinal cord injury. CNS Neurosci Ther 2018; 24(5): 429-38.
[http://dx.doi.org/10.1111/cns.12801] [PMID: 29352519]
[23]
Lukovic D, Stojkovic M, Moreno-Manzano V, et al. Concise review: Reactive astrocytes and stem cells in spinal cord injury: Good guys or bad guys? Stem Cells 2015; 33(4): 1036-41.
[http://dx.doi.org/10.1002/stem.1959] [PMID: 25728093]
[24]
Li X, Li M, Tian L, Chen J, Liu R, Ning B. Reactive astrogliosis: Implications in spinal cord injury progression and therapy. Oxid Med Cell Longev 2020; 2020: 9494352.
[http://dx.doi.org/10.1155/2020/9494352] [PMID: 32884625]
[25]
Yoon C, Tuszynski MH. Frontiers of spinal cord and spine repair: Experimental approaches for repair of spinal cord injury. Adv Exp Med Biol 2012; 760: 1-15.
[http://dx.doi.org/10.1007/978-1-4614-4090-1_1] [PMID: 23281510]
[26]
Alexander JK, Popovich PG. Neuroinflammation in spinal cord injury: Therapeutic targets for neuroprotection and regeneration. Prog Brain Res 2009; 175: 125-37.
[http://dx.doi.org/10.1016/S0079-6123(09)17508-8] [PMID: 19660652]
[27]
Liu H, Zhang J, Xu X, et al. SARM1 promotes neuroinflammation and inhibits neural regeneration after spinal cord injury through NF-κB signaling. Theranostics 2021; 11(9): 4187-206.
[http://dx.doi.org/10.7150/thno.49054] [PMID: 33754056]
[28]
Mulica P, Grünewald A, Pereira SL. Astrocyte-neuron metabolic crosstalk in neurodegeneration: A mitochondrial perspective. Front Endocrinol (Lausanne) 2021; 12: 668517.
[http://dx.doi.org/10.3389/fendo.2021.668517] [PMID: 34025580]
[29]
De Biase D, Valente V, Conte A, et al. Phenotypic effects of homeodomain-interacting protein kinase 2 deletion in mice. Int J Mol Sci 2021; 22(15): 8294.
[http://dx.doi.org/10.3390/ijms22158294] [PMID: 34361060]
[30]
Sims NR, Yew WP. Reactive astrogliosis in stroke: Contributions of astrocytes to recovery of neurological function. Neurochem Int 2017; 107: 88-103.
[http://dx.doi.org/10.1016/j.neuint.2016.12.016] [PMID: 28057555]
[31]
Yu G, Zhang Y, Ning B. Reactive astrocytes in central nervous system injury: Subgroup and potential therapy. Front Cell Neurosci 2021; 15: 792764.
[http://dx.doi.org/10.3389/fncel.2021.792764] [PMID: 35002629]
[32]
Lee KH, Yoon DH, Park YG, Lee BH. Effects of glial transplantation on functional recovery following acute spinal cord injury. J Neurotrauma 2005; 22(5): 575-89.
[http://dx.doi.org/10.1089/neu.2005.22.575] [PMID: 15892602]
[33]
Davies SJ, Shih CH, Noble M, Mayer-Proschel M, Davies JE, Proschel C. Transplantation of specific human astrocytes promotes functional recovery after spinal cord injury. PLoS One 2011; 6(3): e17328.
[http://dx.doi.org/10.1371/journal.pone.0017328] [PMID: 21407803]
[34]
Tan X, Tang H, Bi J, Li N, Jia Y. MicroRNA-222-3p associated with Helicobacter pylori targets HIPK2 to promote cell proliferation, invasion, and inhibits apoptosis in gastric cancer. J Cell Biochem 2018; 119(7): 5153-62.
[http://dx.doi.org/10.1002/jcb.26542] [PMID: 29227536]
[35]
Zhou Y, Li S, Chen P, et al. MicroRNA-27b-3p inhibits apoptosis of chondrocyte in rheumatoid arthritis by targeting HIPK2. Artif Cells Nanomed Biotechnol 2019; 47(1): 1766-71.
[http://dx.doi.org/10.1080/21691401.2019.1607362] [PMID: 31066587]
[36]
Zou F, Xu J, Fu H, et al. Different functions of HIPK2 and CtBP2 in traumatic brain injury. J Mol Neurosci 2013; 49(2): 395-408.
[http://dx.doi.org/10.1007/s12031-012-9906-2] [PMID: 23076816]
[37]
Huang R, Zhang Y, Han B, et al. Circular RNA HIPK2 regulates astrocyte activation via cooperation of autophagy and ER stress by targeting MIR124-2HG. Autophagy 2017; 13(10): 1722-41.
[http://dx.doi.org/10.1080/15548627.2017.1356975] [PMID: 28786753]
[38]
Gao R, Ren L, Zhou Y, et al. Recurrent non-severe hypoglycemia aggravates cognitive decline in diabetes and induces mitochondrial dysfunction in cultured astrocytes. Mol Cell Endocrinol 2021; 526: 111192.
[http://dx.doi.org/10.1016/j.mce.2021.111192] [PMID: 33545179]
[39]
Falnikar A, Li K, Lepore AC. Therapeutically targeting astrocytes with stem and progenitor cell transplantation following traumatic spinal cord injury. Brain Res 2015; 1619: 91-103.
[http://dx.doi.org/10.1016/j.brainres.2014.09.037] [PMID: 25251595]
[40]
Wang SJH, Sinclair DAR, Kim HY, et al. Homeodomain-interacting protein kinase (Hipk) plays roles in nervous system and muscle structure and function. PLoS One 2020; 15(3): e0221006.
[http://dx.doi.org/10.1371/journal.pone.0221006] [PMID: 32187190]
[41]
Sakamoto K, Huang BW, Iwasaki K, Hailemariam K, Ninomiya-Tsuji J, Tsuji Y. Regulation of genotoxic stress response by homeodomain-interacting protein kinase 2 through phosphorylation of cyclic AMP response element-binding protein at serine 271. Mol Biol Cell 2010; 21(16): 2966-74.
[http://dx.doi.org/10.1091/mbc.e10-01-0015] [PMID: 20573984]
[42]
Kim JH, Kim SH, Cho SR, et al. The modulation of neurotrophin and epigenetic regulators: Implication for astrocyte proliferation and neuronal cell apoptosis after spinal cord injury. Ann Rehabil Med 2016; 40(4): 559-67.
[http://dx.doi.org/10.5535/arm.2016.40.4.559] [PMID: 27606261]
[43]
Liu Z, Chopp M. Astrocytes, therapeutic targets for neuroprotection and neurorestoration in ischemic stroke. Prog Neurobiol 2016; 144: 103-20.
[http://dx.doi.org/10.1016/j.pneurobio.2015.09.008] [PMID: 26455456]
[44]
Zhao CL, Cui HA, Zhang XR. MiR-543-5p inhibits inflammation and promotes nerve regeneration through inactivation of the NF-κB in rats after spinal cord injury. Eur Rev Med Pharmacol Sci 2019; 23(3) (Suppl.): 39-46.
[PMID: 31389572]
[45]
Linnerbauer M, Rothhammer V. Protective functions of reactive astrocytes following central nervous system insult. Front Immunol 2020; 11: 573256.
[http://dx.doi.org/10.3389/fimmu.2020.573256] [PMID: 33117368]
[46]
Mancini F, Pieroni L, Monteleone V, et al. MDM4/HIPK2/p53 cytoplasmic assembly uncovers coordinated repression of molecules with anti-apoptotic activity during early DNA damage response. Oncogene 2016; 35(2): 228-40.
[http://dx.doi.org/10.1038/onc.2015.76] [PMID: 25961923]
[47]
Song C, Wu YS, Yang ZY, et al. Astrocyte-conditioned medium protects prefrontal cortical neurons from glutamate-induced cell death by inhibiting TNF-α expression. Neuroimmunomodulation 2019; 26(1): 33-42.
[http://dx.doi.org/10.1159/000495211] [PMID: 30699428]
[48]
Zhang J, Pho V, Bonasera SJ, et al. Essential function of HIPK2 in TGFbeta-dependent survival of midbrain dopamine neurons. Nat Neurosci 2007; 10(1): 77-86.
[http://dx.doi.org/10.1038/nn1816] [PMID: 17159989]
[49]
Zhou Z, Liu C, Chen S, et al. Activation of the Nrf2/ARE signaling pathway by probucol contributes to inhibiting inflammation and neuronal apoptosis after spinal cord injury. Oncotarget 2017; 8(32): 52078-93.
[http://dx.doi.org/10.18632/oncotarget.19107] [PMID: 28881715]
[50]
Torrente L, Sanchez C, Moreno R, et al. Crosstalk between NRF2 and HIPK2 shapes cytoprotective responses. Oncogene 2017; 36(44): 6204-12.
[http://dx.doi.org/10.1038/onc.2017.221] [PMID: 28692050]
[51]
Li D, Tian H, Li X, et al. Zinc promotes functional recovery after spinal cord injury by activating Nrf2/HO-1 defense pathway and inhibiting inflammation of NLRP3 in nerve cells. Life Sci 2020; 245: 117351.
[http://dx.doi.org/10.1016/j.lfs.2020.117351] [PMID: 31981629]
[52]
Liu L, Zhao Z, Yin Q, Zhang X. TTB protects astrocytes against oxygen-glucose deprivation/reoxygenation-induced injury via activation of Nrf2/HO-1 signaling pathway. Front Pharmacol 2019; 10: 792.
[http://dx.doi.org/10.3389/fphar.2019.00792] [PMID: 31379570]