Inhibition of EPAC2 Attenuates Intracerebral Hemorrhage-Induced Secondary Brain Injury via the p38/BIM/Caspase-3 Pathway
- 23 Downloads
Exchange proteins directly activated by cAMP (EPACs) are critical cAMP-dependent signaling pathway intermediaries that have been implicated in the pathogenesis of several human diseases, particularly neurological disorders. However, their pathogenic role in secondary brain injury (SBI) induced by intracranial hemorrhage (ICH) is unknown. The aim of this study was to examine the effects of EPAC2 on ICH-induced SBI and its underlying mechanisms. An in vivo ICH model was established in Sprague–Dawley rats by autologous blood injection. In addition, rat primary cortical neuronal cultures were exposed to oxyhemoglobin to simulate ICH in vitro. The function of EPAC2 in SBI induced by ICH was studied using the EPAC2-specific inhibitor ESI-05. In this study, we found that EPAC2 protein expression was significantly increased in the ICH models in vitro and in vivo. Furthermore, EPAC2 activation was inhibited by ESI-05 under ICH conditions. Inhibition of EPAC2 decreased the apoptosis rate of nerve cells in the cortex accompanied by a corresponding decrease in the protein expression of phosphorylated p38, Bcl-2-like protein 11 (BIM), and caspase-3. In summary, this study showed that inhibition of EPAC2 activation by ESI-05 suppressed SBI induced by ICH via the p38/BIM/caspase-3 signaling pathway.
KeywordsIntracranial hemorrhage Secondary brain injury EPAC2 Apoptosis BIM p38
secondary brain injury
exchange proteins directly activated by cAMP
terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling
This work was supported by the Project of Jiangsu Provincial Medical Innovation Team (No. CXTDA2017003), Suzhou Key Medical Center (No. Szzx201501), Scientific Department of Jiangsu Province (No. BE2017656), Natural Science Foundation of Jiangsu Province (No. BK20180204), Suzhou Government (No. LCZX201601), and grants from the National Natural Science Foundation of China (No. 81571121, No. 81801151 and No. 81771252).
Compliance with Ethical Standards
All experiments were approved by the Ethics Committee of the First Affiliated Hospital of Soochow University (Suzhou, China). The experimental procedures were conducted strictly according to the Animal Health Management Committee of Soochow University. In accordance with their animal ethics criteria, the sample size of this study was determined by efficacy analysis.
Conflict of Interest
The authors declare that there are no conflicts of interest.
- Hua Y, Keep RF, Hoff JT, Xi G (2007) Brain injury after intracerebral hemorrhage: the role of thrombin and iron. Stroke 38:759–762. https://doi.org/10.1161/01.STR.0000247868.97078.10 CrossRefPubMedGoogle Scholar
- Kaufmann T, Jost PJ, Pellegrini M, Puthalakath H, Gugasyan R, Gerondakis S, Cretney E, Smyth MJ, Silke J, Hakem R, Bouillet P, Mak TW, Dixit VM, Strasser A (2009) Fatal hepatitis mediated by tumor necrosis factor TNFα requires caspase-8 and involves the BH3-only proteins Bid and Bim. Immunity 30:56–66CrossRefGoogle Scholar
- Kelly MP, Stein JM, Vecsey CG, Favilla C, Yang X, Bizily SF, Esposito MF, Wand G, Kanes SJ, Abel T (2009) Developmental etiology for neuroanatomical and cognitive deficits in mice overexpressing Galphas, a G-protein subunit genetically linked to schizophrenia. Mol Psychiatry 14:398–415, 347. https://doi.org/10.1038/mp.2008.124 CrossRefPubMedGoogle Scholar
- Kim CG, Castro-Aceituno V, Abbai R, Lee HA, Simu SY, Han Y, Hurh J, Kim YJ, Yang DC (2018) Caspase-3/MAPK pathways as main regulators of the apoptotic effect of the phyto-mediated synthesized silver nanoparticle from dried stem of Eleutherococcus senticosus in human cancer cells. Biomed Pharmacother 99:128–133CrossRefGoogle Scholar
- Matsuda M, Oh-Hashi K, Yokota I, Sawa T, Amaya F (2017) Acquired exchange protein directly activated by cyclic adenosine monophosphate activity induced by p38 mitogen-activated protein kinase in primary afferent neurons contributes to sustaining postincisional nociception. Anesthesiology 126:150–162CrossRefGoogle Scholar
- Middeldorp CM, Vink JM, Hettema JM, de Geus EJC, Kendler KS, Willemsen G, Neale MC, Boomsma DI, Chen X (2010) An association between Epac-1 gene variants and anxiety and depression in two independent samples. Am J Med Genet B Neuropsychiatr Genet 153B:214–219. https://doi.org/10.1002/ajmg.b.30976 CrossRefPubMedPubMedCentralGoogle Scholar
- Pandey M, Varghese M, Sindhu KM, Sreetama S, Navneet A, Mohanakumar KP, Usha R (2008) Mitochondrial NAD+-linked state 3 respiration and complex-I activity are compromised in the cerebral cortex of 3-nitropropionic acid-induced rat model of Huntington’s disease. J Neurochem 104:420–434PubMedGoogle Scholar
- Srivastava DP, Woolfrey KM, Jones KA, Anderson CT, Smith KR, Russell TA, Lee H, Yasvoina MV, Wokosin DL, Ozdinler PH, Shepherd GM, Penzes P (2012b) An autism-associated variant of Epac2 reveals a role for Ras/Epac2 signaling in controlling basal dendrite maintenance in mice. PLoS Biol 10:e1001350. https://doi.org/10.1371/journal.pbio.1001350 CrossRefPubMedPubMedCentralGoogle Scholar
- van Asch CJ, Luitse MJ, Rinkel GJ, van der Tweel I, Algra A, Klijn CJ (2010) Incidence, case fatality, and functional outcome of intracerebral haemorrhage over time, according to age, sex, and ethnic origin: a systematic review and meta-analysis. Lancet Neurol 9:167–176. https://doi.org/10.1016/S1474-4422(09)70340-0 CrossRefPubMedGoogle Scholar