Neuregulin-1β Plays a Neuroprotective Role by Inhibiting the Cdk5 Signaling Pathway after Cerebral Ischemia-Reperfusion Injury in Rats
- 143 Downloads
This study investigated the effects of neuregulin-1β (NRG1β) after middle cerebral artery occlusion/reperfusion (MCAO/R) in rats to evaluate whether they occur via the cyclin-dependent kinase (Cdk)5 signaling pathway. One hundred adult male Wistar rats were randomly divided into sham, MCAO/R, treatment (NRG1β), inhibitor (roscovitine; Ros), and inhibitor + treatment (Ros + NRG1β) groups. The MCAO/R model was established using the intraluminal thread method. The neurobehavioral function was evaluated by the modified neurological severity score (mNSS). The cerebral infarction volume (CIV) was measured by triphenyl tetrazolium chloride (TTC) staining. Morphological changes were observed by hematoxylin-eosin (HE) staining. The apoptotic cell index (ACI) was detected by the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. Immunohistochemistry and Western blotting were performed to detect the expression of calpain 1, p35/p25 (regulatory binding partners of Cdk5), Cdk5, and p-Tau in neurons. The neuronal morphology in the MCAO/R, NRG1β, Ros + NRG1β, and Ros groups differed compared to the sham group; the mNSS, CIV, ACI, and the expression of calpain 1, p35/p25, Cdk5, and p-Tau were significantly increased in all four groups (P < 0.05). In the NRG1β, Ros and Ros + NRG1β groups, the neuronal morphology was significantly improved compared to the MCAO/R group, as were the mNSS, CIV, and ACI. The levels of calpain 1, p35/p25, and p-Tau were decreased compared with the MCAO/R group (P < 0.05), while the Cdk5 expression was not significantly different (P > 0.05). NRG1β may exert neuroprotective effects by inhibiting the expression of calpain 1, p35/p25, and p-Tau after cerebral ischemia-reperfusion injury.
KeywordsCerebral ischemia Reperfusion injury Cdk5 Neuregulin-1β Signaling pathway
This work was supported by grants from the National Natural Science Foundation of China (81274116), the Taishan Scholars Construction Project Excellent Innovative Team of Shandong Province, and the Youth Fund Project of the Affiliated Hospital of Qingdao University. None of these organizations was involved in the design, performance, collection of data, analysis, interpretation, or presentation of this study.
Compliance with Ethical Standards
The National Institutes of Health Guide for the Care and Use of Laboratory Animals was used as a guide for the design of all animal-related studies. All animal-related experiments were performed using protocols approved by our institute (The Ethics Committee of Qingdao University Medical College, QUMC 2011-09).
Conflicts of Interest
The authors declare that they have no conflicts of interest.
- Czapski GA, Gąssowska M, Songin M, Radecka UD, Strosznajder JB (2011) Alterations of cyclin dependent kinase 5 expression and phosphorylation in amyloid precursor protein (APP)-transfected PC12 cells. FEBS Lett 585:1243–1248. https://doi.org/10.1016/j.febslet.2011.03.058 CrossRefPubMedGoogle Scholar
- Fenster C, Vullhorst D, Buonanno A (2012) Acute neuregulin-1 signaling influences AMPA receptor mediated responses in cultured cerebellar granule neurons. Brain Res Bull 87:21–29. https://doi.org/10.1016/j.brainresbull.2011.10.011 CrossRefPubMedGoogle Scholar
- Guo YL, Gao HM, Li ZX, Liu XJ, Du F, Zhang R, Li Q, Wang T (2008) The evaluation of focal cerebral ischemic reperfusion model with middle cerebral artery occlusion by micro-balloon in rhesus monkeys. Acta Anatomica Sinica 39:944–947Google Scholar
- Huang YZ, Mei L (2001) Signal transduction of neuregulin/ErbB in the development of nervous system. Prog Physiol Sci 32:197–203Google Scholar
- Ji YQ, Zhang R, Teng L, Li HY, Guo YL (2017) Study of neuron-protective effect and mechanism of neuregulin1β against cerebral ischemia reperfusion-induced injury in rats. Natl Med J China 97:2128–2134. https://doi.org/10.3760/cma.j.issn.0376-2491.2017.27.011 CrossRefGoogle Scholar
- Li BS, Ma W, Jaffe H, Zheng Y, Takahashi S, Zhang L, Kulkarni AB, Pant HC (2003) Cyclin-dependent kinase-5 is involved in neuregulin-dependent activation of phosphatidylinositol 3-kinase and Akt activity mediating neuronal survival. J Biol Chem 278:35702–35709. https://doi.org/10.1074/jbc.M302004200 CrossRefPubMedGoogle Scholar
- Li Y, Lein PJ, Liu C, Bruun DA, Giulivi C, Ford GD, Tewolde T, Ross-Inta C, Ford BD (2012) Neuregulin-1 is neuroprotective in a rat model of organophosphate-induced delayed neuronal injury. Toxicol Appl Pharmacol 262:194–204. https://doi.org/10.1016/j.taap.2012.05.001 CrossRefPubMedPubMedCentralGoogle Scholar
- Menn B, Bach S, Blevins TL, Campbell M, Meijer L, Timsit S (2010) Delayed treatment with systemic (S)-roscovitine provides neuroprotection and inhibits in vivo CDK5 activity increase in animal stroke models. PLoS One 5:e12117. https://doi.org/10.1371/journal.pone.0012117 CrossRefPubMedPubMedCentralGoogle Scholar
- Otth C, Mendoza-Naranjo A, Mujica L, Zambrano A, Concha II, Maccioni RB (2003) Modulation of the JNK and p38 pathways by cdk5 protein kinase in a transgenic mouse model of Alzheimer’s disease. Neuroreport 14:2403–2409. https://doi.org/10.1097/01.wnr.0000099988.54721.3c CrossRefPubMedGoogle Scholar
- Qu D, Rashidian J, Mount MP, Aleyasin H, Parsanejad M, Lira A, Haque E, Zhang Y, Callaghan S, Daigle M, Rousseaux MW, Slack RS, Albert PR, Vincent I, Woulfe JM, Park DS (2007) Role of Cdk5-mediated phosphorylation of Prx2 in MPTP toxicity and Parkinson’s disease. Neuron 55:37–52. https://doi.org/10.1016/j.neuron.2007.05.033 CrossRefPubMedGoogle Scholar
- Rai A, Tripathi S, Kushwaha R, Singh P, Srivastava P, Sanyal S, Bandyopadhyay S (2014) CDK5-induced p-PPARγ(Ser 112) downregulates GFAP via PPREs in developing rat brain: effect of metal mixture and troglitazone in astrocytes. Cell Death Dis 5:e1033. https://doi.org/10.1038/cddis.2013.514 CrossRefPubMedPubMedCentralGoogle Scholar
- Rashidian J, Rousseaux MW, Venderova K, Qu D, Callaghan SM, Phillips M, Bland RJ, During MJ, Mao Z, Slack RS, Park DS (2009) Essential role of cytoplasmic cdk5 and Prx2 in multiple ischemic injury models, in vivo. J Neurosci 29:12497–12505. https://doi.org/10.1523/JNEUROSCI.3892-09.2009 CrossRefPubMedPubMedCentralGoogle Scholar
- Rui Z, Qin L, Wang T, Fang D, Guo YL (2009) Interfering effect of neuregulin upon cerebral ischemic reperfusion injury in rhesus monkeys. Natl Med J China 89:1361–1364Google Scholar
- Tan X, Chen Y, Li J, Li X, Miao Z, Xin N, Zhu J, Ge W, Feng Y, Xu X (2015) The inhibition of Cdk5 activity after hypoxia/ischemia injury reduces infarct size and promotes functional recovery in neonatal rats. Neurosci 290:552–560. https://doi.org/10.1016/j.neuroscience.2015.01.054 CrossRefGoogle Scholar
- Woo RS, Lee JH, Kim HS, Baek CH, Song DY, Suh YH, Baik TK (2012) Neuregulin-1 protects against neurotoxicities induced by Swedish amyloid precursor protein via the Erbb4 receptor. Neurosci 202:413–423. https://doi.org/10.1016/j.neuroscience.2011.11.026 CrossRefGoogle Scholar
- Xu Z, Ford GD, Croslan DR, Jiang J, Gates A, Allen R, Ford BD (2005) Neuroprotection by neuregulin-1 following focal stroke is associated with the attenuation of ischemia-induced pro-inflammatory and stress gene expression. Neurobiol Dis 19:461–470. https://doi.org/10.1016/j.nbd.2005.01.027 CrossRefPubMedGoogle Scholar