Generic placeholder image

Current Neurovascular Research

Editor-in-Chief

ISSN (Print): 1567-2026
ISSN (Online): 1875-5739

Research Article

Peptide5 Attenuates rtPA Related Brain Microvascular Endothelial Cells Reperfusion Injury via the Wnt/β-Catenin Signalling Pathway

Author(s): Weimin Ren, Chuyi Huang, Heling Chu, Yuping Tang and Xiaobo Yang*

Volume 18, Issue 2, 2021

Published on: 09 August, 2021

Page: [219 - 226] Pages: 8

DOI: 10.2174/1567202618666210809115305

Price: $65

conference banner
Abstract

Aims: Brain vascular endothelial cell dysfunction after rtPA treatment is a significant factor associated with poor prognosis, suggesting that alleviation of rtPA-related endothelial cell injury may represent a potential beneficial strategy along with rtPA thrombolysis.

Background: Thrombolysis with recombinant tissue plasminogen activator (rtPA) is beneficial for acute ischemic stroke but may increase the risk of Hemorrhagic Transformation (HT), which is considered ischemia-reperfusion injury. The underlying reason may contribute to brain endothelial injury and dysfunction related to rtPA against ischemic stroke. As previous studies have demonstrated that transiently blocked Cx43 using peptide5 (Cx43 mimetic peptide) during retinal ischemia reduced vascular leakage, it is necessary to know whether this might help decrease side effect of rtPA within the therapeutic time window.

Objective: This study aims to investigate the effects of peptide5 on rtPA-related cell injury during hypoxia/reoxygenation (H/R) within the therapeutic time window.

Methods: In this study, we established a cell hypoxia/reoxygenation H/R model in cultured primary Rat Brain Microvascular Endothelial Cells (RBMECs) and evaluated endothelial cell death and permeability after rtPA treatment with or without transient peptide5. In addition, we also investigated the potential signaling pathway to explore the underlying mechanisms preliminarily.

Results: The results showed that peptide5 inhibited rtPA-related endothelial cell death and permeability. It also slightly increased tight junction (ZO-1, occluding, claudin-5) and β-catenin mRNA expression, demonstrating that peptide5 might attenuate endothelial cell injury by regulating the Wnt/ β-catenin pathway. The following bioinformatic exploration from the GEO dataset GSE37239 was also consistent with our findings.

Conclusion: This study showed that the application of peptide5 maintained cell viability and permeability associated with rtPA treatment, revealing a possible pathway that could be exploited to limit rtPA-related endothelial cell injury during ischemic stroke. Furthermore, the altered Wnt/β- catenin signaling pathway demonstrated that signaling pathways associated with Cx43 might have potential applications in the future. This study may provide a new way to attenuate HT and assist the application of rtPA in ischemic stroke.

Keywords: rtPA, connexin43, mimetic peptide, microvascular endothelial cells, ischemic stroke, signalling pathway.

[1]
Zivin JA. Acute stroke therapy with tissue plasminogen activator (tPA) since it was approved by the U.S. Food and Drug Administration (FDA). Ann Neurol 2009; 66(1): 6-10.
[http://dx.doi.org/10.1002/ana.21750] [PMID: 19681102]
[2]
Pande SD, Win MM, Khine AA, et al. Haemorrhagic transformation following ischaemic stroke: A retrospective study. Sci Rep 2020; 10(1): 5319.
[http://dx.doi.org/10.1038/s41598-020-62230-5] [PMID: 32210323]
[3]
Lansberg MG, Albers GW, Wijman CA. Symptomatic intracerebral hemorrhage following thrombolytic therapy for acute ischemic stroke: A review of the risk factors. Cerebrovasc Dis 2007; 24(1): 1-10.
[http://dx.doi.org/10.1159/000103110] [PMID: 17519538]
[4]
Hacke W, Kaste M, Bluhmki E, et al. Thrombolysis with alteplase 3 to 4.5 hours after acute ischemic stroke. N Engl J Med 2008; 359(13): 1317-29.
[http://dx.doi.org/10.1056/NEJMoa0804656] [PMID: 18815396]
[5]
Jensen M, Schlemm E, Cheng B, et al. Clinical characteristics and outcome of patients with hemorrhagic transformation after intravenous thrombolysis in the WAKE-UP trial. Front Neurol 2020; 11: 957.
[http://dx.doi.org/10.3389/fneur.2020.00957] [PMID: 32982951]
[6]
Kenna JE, Anderton RS, Knuckey NW, Meloni BP. Assessment of recombinant tissue plasminogen activator (rtPA) toxicity in cultured neural cells and subsequent treatment with poly-arginine peptide R18D. Neurochem Res 2020; 45(5): 1215-29.
[http://dx.doi.org/10.1007/s11064-020-03004-3] [PMID: 32140956]
[7]
Danesh-Meyer HV, Kerr NM, Zhang J, et al. Connexin43 mimetic peptide reduces vascular leak and retinal ganglion cell death following retinal ischaemia. Brain 2012; 135(Pt 2): 506-20.
[http://dx.doi.org/10.1093/brain/awr338] [PMID: 22345088]
[8]
Chong ZZ, Shang YC, Maiese K. Vascular injury during elevated glucose can be mitigated by erythropoietin and Wnt signaling. Curr Neurovasc Res 2007; 4(3): 194-204.
[http://dx.doi.org/10.2174/156720207781387150] [PMID: 17691973]
[9]
Wang L, Fan W, Cai P, et al. Recombinant ADAMTS13 reduces tissue plasminogen activator-induced hemorrhage after stroke in mice. Ann Neurol 2013; 73(2): 189-98.
[http://dx.doi.org/10.1002/ana.23762] [PMID: 23280993]
[10]
Miyazaki T, Kimura Y, Ohata H, et al. Distinct effects of tissue- type plasminogen activator and SMTP-7 on cerebrovascular inflammation following thrombolytic reperfusion. Stroke 2011; 42(4): 1097-104.
[http://dx.doi.org/10.1161/STROKEAHA.110.598359] [PMID: 21350203]
[11]
O’Carroll SJ, Gorrie CA, Velamoor S, Green CR, Nicholson LF. Connexin43 mimetic peptide is neuroprotective and improves function following spinal cord injury. Neurosci Res 2013; 75(3): 256-67.
[http://dx.doi.org/10.1016/j.neures.2013.01.004] [PMID: 23403365]
[12]
Mao Y, Tonkin RS, Nguyen T, et al. Systemic administration of Connexin43 mimetic peptide improves functional recovery after traumatic spinal cord injury in adult rats. J Neurotrauma 2017; 34(3): 707-19.
[http://dx.doi.org/10.1089/neu.2016.4625] [PMID: 27629792]
[13]
Bao Dang Q, Lapergue B, Tran-Dinh A, et al. High-density lipoproteins limit neutrophil-induced damage to the blood-brain barrier in vitro. J Cereb Blood Flow Metab 2013; 33(4): 575-82.
[http://dx.doi.org/10.1038/jcbfm.2012.206] [PMID: 23299241]
[14]
Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 2008; 3(6): 1101-8.
[http://dx.doi.org/10.1038/nprot.2008.73] [PMID: 18546601]
[15]
Davis S, Meltzer PS. GEOquery: A bridge between the Gene Expression Omnibus (GEO) and BioConductor. Bioinformatics 2007; 23(14): 1846-7.
[http://dx.doi.org/10.1093/bioinformatics/btm254] [PMID: 17496320]
[16]
Huang W, Sherman BT, Lempicki RA. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 2009; 4(1): 44-57.
[http://dx.doi.org/10.1038/nprot.2008.211] [PMID: 19131956]
[17]
Garraud M, Khacef K, Vion AC, et al. Recombinant tissue plasminogen activator enhances microparticle release from mouse brain-derived endothelial cells through plasmin. J Neurol Sci 2016; 370: 187-95.
[http://dx.doi.org/10.1016/j.jns.2016.09.026] [PMID: 27772757]
[18]
Lo EH, Dalkara T, Moskowitz MA. Mechanisms, challenges and opportunities in stroke. Nat Rev Neurosci 2003; 4(5): 399-415.
[http://dx.doi.org/10.1038/nrn1106] [PMID: 12728267]
[19]
Hoorelbeke D, Decrock E, De Smet M, et al. Cx43 channels and signaling via IP3/Ca2+, ATP, and ROS/NO propagate radiation-induced DNA damage to non-irradiated brain microvascular endothelial cells. Cell Death Dis 2020; 11(3): 194.
[http://dx.doi.org/10.1038/s41419-020-2392-5] [PMID: 32188841]
[20]
Morioka N, Kondo S, Harada N, et al. Downregulation of connexin43 potentiates noradrenaline-induced expression of brain-derived neurotrophic factor in primary cultured cortical astrocytes. J Cell Physiol 2021; 236(10): 6777-92.
[http://dx.doi.org/10.1002/jcp.30353] [PMID: 33665818]
[21]
De Bock M, Wang N, Decrock E, et al. Endothelial calcium dynamics, connexin channels and blood-brain barrier function. Prog Neurobiol 2013; 108: 1-20.
[http://dx.doi.org/10.1016/j.pneurobio.2013.06.001] [PMID: 23851106]
[22]
Wang N, De Bock M, Antoons G, et al. Connexin mimetic peptides inhibit Cx43 hemichannel opening triggered by voltage and intracellular Ca2+ elevation. Basic Res Cardiol 2012; 107(6): 304.
[http://dx.doi.org/10.1007/s00395-012-0304-2] [PMID: 23095853]
[23]
Abudara V, Bechberger J, Freitas-Andrade M, et al. The connexin43 mimetic peptide Gap19 inhibits hemichannels without altering gap junctional communication in astrocytes. Front Cell Neurosci 2014; 8: 306.
[http://dx.doi.org/10.3389/fncel.2014.00306] [PMID: 25374505]
[24]
Wang N, De Vuyst E, Ponsaerts R, et al. Selective inhibition of Cx43 hemichannels by Gap19 and its impact on myocardial ischemia/reperfusion injury. Basic Res Cardiol 2013; 108(1): 309.
[http://dx.doi.org/10.1007/s00395-012-0309-x] [PMID: 23184389]
[25]
Kim Y, Griffin JM, Harris PW, et al. Characterizing the mode of action of extracellular Connexin43 channel blocking mimetic peptides in an in vitro ischemia injury model. Biochim Biophys Acta, Gen Subj 2017; 1861(2): 68-78.
[http://dx.doi.org/10.1016/j.bbagen.2016.11.001] [PMID: 27816754]
[26]
Mugisho OO, Green CR, Squirrell DM, et al. Connexin43 hemichannel block protects against the development of diabetic retinopathy signs in a mouse model of the disease. J Mol Med (Berl) 2019; 97(2): 215-29.
[http://dx.doi.org/10.1007/s00109-018-1727-5] [PMID: 30535867]
[27]
Tonkin RS, Bowles C, Perera CJ, et al. Attenuation of mechanical pain hypersensitivity by treatment with Peptide5, a connexin-43 mimetic peptide, involves inhibition of NLRP3 inflammasome in nerve-injured mice. Exp Neurol 2018; 300: 1-12.
[http://dx.doi.org/10.1016/j.expneurol.2017.10.016] [PMID: 29055716]
[28]
Guo CX, Mat Nor MN, Danesh-Meyer HV, et al. Connexin43 mimetic peptide improves retinal function and reduces inflammation in a light-damaged albino rat model. Invest Ophthalmol Vis Sci 2016; 57(10): 3961-73.
[http://dx.doi.org/10.1167/iovs.15-16643] [PMID: 27490318]
[29]
Ramadan R, Vromans E, Anang DC, et al. Connexin43 hemichannel targeting with TAT-Gap19 alleviates radiation-induced endothelial cell damage. Front Pharmacol 2020; 11: 212.
[http://dx.doi.org/10.3389/fphar.2020.00212] [PMID: 32210810]
[30]
Zhang L, Xu S, Wu X, et al. Combined treatment with 2-(2-Benzofu-Ranyl)-2-imidazoline and recombinant tissue plasminogen activator protects blood-brain barrier integrity in a rat model of embolic middle cerebral artery occlusion. Front Pharmacol 2020; 11: 801.
[http://dx.doi.org/10.3389/fphar.2020.00801] [PMID: 32595494]
[31]
Liberale L, Bertolotto M, Minetti S, et al. Recombinant tissue plasminogen activator (r-tpa) induces in-vitro human neutrophil migration via low density Lipoprotein Receptor-Related Protein 1 (LRP-1). Int J Mol Sci 2020; 21(19): E7014.
[http://dx.doi.org/10.3390/ijms21197014] [PMID: 32977685]
[32]
Saleem S, Wang D, Zhao T, Sullivan RD, Reed GL. Matrix metalloproteinase-9 expression is enhanced by ischemia and tissue plasminogen activator and induces hemorrhage, disability and mortality in experimental stroke. Neuroscience 2021; 460: 120-9.
[http://dx.doi.org/10.1016/j.neuroscience.2021.01.003] [PMID: 33465414]
[33]
Dhanda S, Sandhir R. Blood-brain barrier permeability is exacerbated in experimental model of hepatic encephalopathy via mmp-9 activation and downregulation of tight junction proteins. Mol Neurobiol 2018; 55(5): 3642-59.
[PMID: 28523565]
[34]
Zhang J, Yang G, Zhu Y, Peng X, Li T, Liu L. Relationship of Cx43 regulation of vascular permeability to osteopontin-tight junction protein pathway after sepsis in rats. Am J Physiol Regul Integr Comp Physiol 2018; 314(1): R1-R11.
[http://dx.doi.org/10.1152/ajpregu.00443.2016] [PMID: 28978514]
[35]
Maiese K. Heightened attention for Wnt signaling in diabetes mellitus. Curr Neurovasc Res 2020; 17(3): 215-7.
[http://dx.doi.org/10.2174/1567202617999200327134835] [PMID: 32216737]
[36]
Lv X, Li J, Hu Y, et al. Overexpression of miR-27b-3p Targeting Wnt3a regulates the signaling pathway of Wnt/β-catenin and attenuates atrial fibrosis in rats with atrial fibrillation. Oxid Med Cell Longev 2019; 2019: 5703764.
[http://dx.doi.org/10.1155/2019/5703764] [PMID: 31178968]
[37]
Rinaldi F, Hartfield EM, Crompton LA, et al. Cross-regulation of Connexin43 and β-catenin influences differentiation of human neural progenitor cells. Cell Death Dis 2014; 5: e1017.
[http://dx.doi.org/10.1038/cddis.2013.546] [PMID: 24457961]
[38]
López C, Aguilar R, Nardocci G, et al. Wnt/β-catenin signaling enhances transcription of the CX43 gene in murine Sertoli cells. J Cell Biochem 2019; 120(4): 6753-62.
[http://dx.doi.org/10.1002/jcb.27973] [PMID: 30417410]
[39]
Ji YB, Gao Q, Tan XX, et al. Lithium alleviates blood-brain barrier breakdown after cerebral ischemia and reperfusion by upregulating endothelial Wnt/β-catenin signaling in mice. Neuropharmacology 2021; 186: 108474.
[http://dx.doi.org/10.1016/j.neuropharm.2021.108474] [PMID: 33524408]

Rights & Permissions Print Cite
© 2024 Bentham Science Publishers | Privacy Policy