CNTF protects neurons from hypoxic injury through the activation of STAT3pTyr705

Gu,Ying  Li; Gao,Guan  Qun; Ma,Ning; Ye,Lin  Lin; Zhang,Li  Wei; Gao,Xu; Zhang,Zhuo  Bo

doi:10.3892/ijmm.2016.2769

CNTF protects neurons from hypoxic injury through the activation of STAT3pTyr705

Authors:
- Ying Li Gu
- Guan Qun Gao
- Ning Ma
- Lin Lin Ye
- Li Wei Zhang
- Xu Gao
- Zhuo Bo Zhang
View Affiliations

Affiliations: Department of Neurology, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China, Department of Biochemistry and Molecular Biology, Harbin Medical University, Harbin, Heilongjiang 150081, P.R. China
Published online on: October 13, 2016 https://doi.org/10.3892/ijmm.2016.2769
Pages: 1915-1921

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of the present study was to investigate whether ciliary neurotrophic factor (CNTF) plays its neuroprotective role following hypoxic injury through the activation of signal transducer and activator of transcription 3 (STAT3) signaling. Firstly, to determine whether CNTF exerts its effects via STAT3 following hypoxic injury, cultured neurons from the cerebral cortex of mice were prepared and a neuronal model of hypoxia was then established. The neurons exposed to hypoxia were then pre-treated with CNTF and transfected with small interference RNA (siRNA) targeting STAT3 (STAT3 siRNA) using polybrene, or with STAT3Tyr705 mutant or STAT3Ser727 mutant using an electroporation system. The survival, proliferation and neurite outgrowth of the neurons subjected to different treatments were also determined. RT-qPCR and western blot analysis were employed to examine the expression levels of STAT3, p-STAT3Tyr705 and p-STAT3Ser727 following treatment with CNTF and other treatments. Our results revealed that treatment with CNTF: i) protected neurons from hypoxic injury by promoting survival and neurite growth; ii) induced a significant increase in the levels of STAT3, STAT3pTyr705 and the STAT3pTyr705/STAT3 ratio; it did not however, significantly affect the levels of STAT3pSer727 in the hypoxic cerebral cortex neurons. Transfection of the hypoxic neurons pre-treated with CNTF with STAT3 siRNA or STAT3Tyr705 neutralized the protective effects exerted by CNTF. The findings of our study thus demonstrate that CNTF protects neurons from hypoxic injury through the activation of STAT3pTyr705.

View Figures

View References

1	du Plessis AJ and Volpe JJ: Perinatal brain injury in the preterm and term newborn. Curr Opin Neurol. 15:151–157. 2002. View Article : Google Scholar : PubMed/NCBI
2	Azra Haider B and Bhutta ZA: Birth asphyxia in developing countries: current status and public health implications. Curr Probl Pediatr Adolesc Health Care. 36:178–188. 2006. View Article : Google Scholar : PubMed/NCBI
3	Jiang H, Lei JJ and Zhang YH: Protective effect of topiramate on hypoxic-ischemic brain injury in neonatal rat. Asian Pac J Trop Med. 7:496–500. 2014. View Article : Google Scholar : PubMed/NCBI
4	Marín-Padilla M: Perinatal brain damage, cortical reorganization (acquired cortical dysplasias), and epilepsy. Adv Neurol. 84:153–172. 2000.PubMed/NCBI
5	Robinson S: Systemic prenatal insults disrupt telencephalon development: implications for potential interventions. Epilepsy Behav. 7:345–363. 2005. View Article : Google Scholar : PubMed/NCBI
6	Volpe JJ: Brain injury in premature infants: a complex amalgam of destructive and developmental disturbances. Lancet Neurol. 8:110–124. 2009. View Article : Google Scholar :
7	Martinez-Biarge M, Diez-Sebastian J, Rutherford MA and Cowan FM: Outcomes after central grey matter injury in term perinatal hypoxic-ischaemic encephalopathy. Early Hum Dev. 86:675–682. 2010. View Article : Google Scholar : PubMed/NCBI
8	Ferriero DM: Neonatal brain injury. N Engl J Med. 351:1985–1995. 2004. View Article : Google Scholar : PubMed/NCBI
9	Juul SE and Ferriero DM: Pharmacologic neuroprotective strategies in neonatal brain injury. Clin Perinatol. 41:119–131. 2014. View Article : Google Scholar : PubMed/NCBI
10	Levene ML, Kornberg J and Williams TH: The incidence and severity of post-asphyxial encephalopathy in full-term infants. Early Hum Dev. 11:21–26. 1985. View Article : Google Scholar : PubMed/NCBI
11	Cerio FG, Lara-Celador I, Álvarez A and Hilario E: Neuroprotective therapies after perinatal hypoxic-ischemic brain injury. Brain Sci. 3:191–214. 2013. View Article : Google Scholar : PubMed/NCBI
12	Skaper SD, Selak I, Manthorpe M and Varon S: Chemically defined requirements for the survival of cultured 8-day chick embryo ciliary ganglion neurons. Brain Res. 302:281–290. 1984. View Article : Google Scholar : PubMed/NCBI
13	Wagener EM, Aurich M, Aparicio-Siegmund S, Floss DM, Garbers C, Breusing K, Rabe B, Schwanbeck R, Grötzinger J, Rose-John S and Scheller J: The amino acid exchange R28E in ciliary neurotrophic factor (CNTF) abrogates interleukin-6 receptor-dependent but retains CNTF receptor-dependent signaling via glycoprotein 130 (gp130)/leukemia inhibitory factor receptor (LIFR). J Biol Chem. 289:18442–18450. 2014. View Article : Google Scholar : PubMed/NCBI
14	Wang K, Zhou F, Zhu X, Zhang K, Huang B and Zhu L and Zhu L: Neuroprotective properties of ciliary neurotrophic factor on retinoic acid (RA)-predifferentiated SH-SY5Y neuroblastoma cells. Folia Neuropathol. 52:121–127. 2014. View Article : Google Scholar : PubMed/NCBI
15	Askvig JM and Watt JA: The MAPK and PI3K pathways mediate CNTF-induced neuronal survival and process outgrowth in hypothalamic organotypic cultures. J Cell Commun Signal. 9:217–231. 2015. View Article : Google Scholar : PubMed/NCBI
16	Zhou Q, Chen P, Di G, Zhang Y, Wang Y, Qi X, Duan H and Xie L: Ciliary neurotrophic factor promotes the activation of corneal epithelial stem/progenitor cells and accelerates corneal epithelial wound healing. Stem Cells. 33:1566–1576. 2015. View Article : Google Scholar
17	Severi I, Senzacqua M, Mondini E, Fazioli F, Cinti S and Giordano A: Activation of transcription factors STAT1 and STAT5 in the mouse median eminence after systemic ciliary neurotrophic factor administration. Brain Res. 1622:217–229. 2015. View Article : Google Scholar : PubMed/NCBI
18	Covey MV and Levison SW: Leukemia inhibitory factor participates in the expansion of neural stem/progenitors after perinatal hypoxia/ischemia. Neuroscience. 148:501–509. 2007. View Article : Google Scholar : PubMed/NCBI
19	Shrivastava K, Llovera G, Recasens M, Chertoff M, Giménez-Llort L, Gonzalez B and Acarin L: Temporal expression of cytokines and signal transducer and activator of transcription factor 3 activation after neonatal hypoxia/ischemia in mice. Dev Neurosci. 35:212–225. 2013. View Article : Google Scholar : PubMed/NCBI
20	Schwieger J, Warnecke A, Lenarz T, Esser KH and Scheper V: Neuronal survival, morphology and outgrowth of spiral ganglion neurons using a defined growth factor combination. PLoS One. 10:e01336802015. View Article : Google Scholar : PubMed/NCBI
21	Lin JC, Xing YL, Xu WM, Li M, Bo P, Niu YY and Zhang CR: Evaluation of galactomannan enzyme immunoassay and quantitative real-time PCR for the diagnosis of invasive pulmonary aspergillosis in a rat model. J Microbiol Biotechnol. 24:1044–1050. 2014. View Article : Google Scholar : PubMed/NCBI
22	Quesnelle KM, Boehm AL and Grandis JR: STAT-mediated EGFR signaling in cancer. J Cell Biochem. 102:311–319. 2007. View Article : Google Scholar : PubMed/NCBI
23	Frank DA: STAT3 as a central mediator of neoplastic cellular transformation. Cancer Lett. 251:199–210. 2007. View Article : Google Scholar
24	Germain D and Frank DA: Targeting the cytoplasmic and nuclear functions of signal transducers and activators of transcription 3 for cancer therapy. Clin Cancer Res. 13:5665–5669. 2007. View Article : Google Scholar : PubMed/NCBI
25	Northington FJ, Chavez-Valdez R and Martin LJ: Neuronal cell death in neonatal hypoxia-ischemia. Ann Neurol. 69:743–758. 2011. View Article : Google Scholar : PubMed/NCBI
26	Li X, Zhang J, Chai S and Wang X: Progesterone alleviates hypoxic-ischemic brain injury via the Akt/GSK-3β signaling pathway. Exp Ther Med. 8:1241–1246. 2014.PubMed/NCBI
27	Linker R, Gold R and Luhder F: Function of neurotrophic factors beyond the nervous system: inflammation and autoimmune demyelination. Crit Rev Immunol. 29:43–68. 2009. View Article : Google Scholar : PubMed/NCBI
28	Maisonpierre PC, Belluscio L, Squinto S, Ip NY, Furth ME, Lindsay RM and Yancopoulos GD: Neurotrophin-3: a neurotrophic factor related to NGF and BDNF. Science. 247:1446–1451. 1990. View Article : Google Scholar : PubMed/NCBI
29	Davis S, Aldrich TH, Valenzuela DM, Wong VV, Furth ME, Squinto SP and Yancopoulos GD: The receptor for ciliary neurotrophic factor. Science. 253:59–63. 1991. View Article : Google Scholar : PubMed/NCBI
30	Ip NY, Nye SH, Boulton TG, Davis S, Taga T, Li Y, Birren SJ, Yasukawa K, Kishimoto T, Anderson DJ, et al: CNTF and LIF act on neuronal cells via shared signaling pathways that involve the IL-6 signal transducing receptor component gp130. Cell. 69:1121–1132. 1992. View Article : Google Scholar : PubMed/NCBI
31	Sleeman MW, Anderson KD, Lambert PD, Yancopoulos GD and Wiegand SJ: The ciliary neurotrophic factor and its receptor, CNTFR alpha. Pharm Acta Helv. 74:265–272. 2000. View Article : Google Scholar : PubMed/NCBI
32	Turnley AM and Bartlett PF: Cytokines that signal through the leukemia inhibitory factor receptor-beta complex in the nervous system. J Neurochem. 74:889–899. 2000. View Article : Google Scholar : PubMed/NCBI
33	Stahl N, Boulton TG, Farruggella T, Ip NY, Davis S, Witthuhn BA, Quelle FW, Silvennoinen O, Barbieri G, Pellegrini S, et al: Association and activation of Jak-Tyk kinases by CNTF-LIF-OSM-IL-6 beta receptor components. Science. 263:92–95. 1994. View Article : Google Scholar : PubMed/NCBI
34	Boulton TG, Stahl N and Yancopoulos GD: Ciliary neurotrophic factor/leukemia inhibitory factor/interleukin 6/oncostatin M family of cytokines induces tyrosine phosphorylation of a common set of proteins overlapping those induced by other cytokines and growth factors. J Biol Chem. 269:11648–11655. 1994.PubMed/NCBI
35	Kaur N, Kim IJ, Higgins D and Halvorsen SW: Induction of an interferon-γ Stat3 response in nerve cells by pre-treatment with gp130 cytokines. J Neurochem. 87:437–447. 2003. View Article : Google Scholar : PubMed/NCBI
36	Kaur N, Wohlhueter AL and Halvorsen SW: Activation and inactivation of signal transducers and activators of transcription by ciliary neurotrophic factor in neuroblastoma cells. Cell Signal. 14:419–429. 2002. View Article : Google Scholar : PubMed/NCBI
37	Alonzi T, Middleton G, Wyatt S, Buchman V, Betz UA, Müller W, Musiani P, Poli V and Davies AM: Role of STAT3 and PI 3-kinase/Akt in mediating the survival actions of cytokines on sensory neurons. Mol Cell Neurosci. 18:270–282. 2001. View Article : Google Scholar : PubMed/NCBI
38	Schweizer U, Gunnersen J, Karch C, Wiese S, Holtmann B, Takeda K, Akira S and Sendtner M: Conditional gene ablation of Stat3 reveals differential signaling requirements for survival of motoneurons during development and after nerve injury in the adult. J Cell Biol. 156:287–297. 2002. View Article : Google Scholar : PubMed/NCBI
39	Zhong Z, Wen Z and Darnell JE Jr: Stat3: a STAT family member activated by tyrosine phosphorylation in response to epidermal growth factor and interleukin-6. Science. 264:95–98. 1994. View Article : Google Scholar : PubMed/NCBI
40	Sango K, Yanagisawa H, Komuta Y, Si Y and Kawano H: Neuroprotective properties of ciliary neurotrophic factor for cultured adult rat dorsal root ganglion neurons. Histochem Cell Biol. 130:669–679. 2008. View Article : Google Scholar : PubMed/NCBI
41	Rhee KD, Goureau O, Chen S and Yang XJ: Cytokine-induced activation of signal transducer and activator of transcription in photoreceptor precursors regulates rod differentiation in the developing mouse retina. J Neurosci. 24:9779–9788. 2004. View Article : Google Scholar : PubMed/NCBI
42	Leeman RJ, Lui VW and Grandis JR: STAT3 as a therapeutic target in head and neck cancer. Expert Opin Biol Ther. 6:231–241. 2006. View Article : Google Scholar : PubMed/NCBI
43	Johnston PA and Grandis JR: STAT3 signaling: anticancer strategies and challenges. Mol Interv. 11:18–26. 2011. View Article : Google Scholar : PubMed/NCBI
44	Regis G, Pensa S, Boselli D, Novelli F and Poli V: Ups and downs: the STAT1:STAT3 seesaw of interferon and gp130 receptor signalling. Semin Cell Dev Biol. 19:351–359. 2008. View Article : Google Scholar : PubMed/NCBI