Skip to main content
SpringerLink
Log in

Quinolinic acid induces cell apoptosis in PC12 cells through HIF-1-dependent RTP801 activation

  • Original Article
  • Published:
Metabolic Brain Disease Aims and scope Submit manuscript

Abstract

Neurological disease comprises a series of disorders featuring brain dysfunction and neuronal cell death. Among the factors contributing to neuronal death, excitotoxicity induced by excitatory amino acids, such as glutamate, plays a critical role. However, the mechanisms about how the excitatory amino acids induce neuronal death remain elucidated. In this study, we investigated the role of HIF-1α (hypoxia inducible factor-1α) and RTP801 in cell apoptosis induced by quinolinic acid (QUIN), a glutamatergic agonist, in PC12 cells. We found that QUIN at 5 μM increased the expression of HIF-1α significantly with a peak at 24 h. After the treatment with QUIN (5–20 μM) for 24 h, the cells exhibited decreased viability and cell apoptosis with a concomitant increased expression of apoptosis related proteins. QUIN treatment also induced the generation of intracellular reactive oxygen species and RTP801 up-regulation in a HIF-1α-dependent manner that were inhibited by 2-methoxyestradiol, a HIF-1α inhibitor. Importantly, HIF-1 or RTP801 invalidation by siRNA rescued the cell apoptosis induced by QUIN or cobalt chloride, a chemical inducer of HIF-1. Taken together, these findings support the concept that neurotoxicity induced by QUIN is associated with HIF-1-dependent RTP801 activation and provide insight into the potential of RTP801 inhibitor in treatment of neurological disorders.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Agrawal M, Kumar V, Kashyap MP, Khanna VK, Randhawa GS, Pant AB (2011) Ischemic insult induced apoptotic changes in PC12 cells: protection by transresveratrol. Eur J Pharmacol 666:5–11

    Article  CAS  PubMed  Google Scholar 

  • Baranova O, Miranda LF, Pichiule P, Dragatsis I, Johnson RS, Chavez JC (2007) Neuron-specific inactivation of the hypoxia inducible factor 1 alpha increases brain injury in a mouse model of transient focal cerebral ischemia. J Neurosci 27:6320–6332

    Article  CAS  PubMed  Google Scholar 

  • Barger SW, Goodwin ME, Porter MM, Beggs ML (2007) Glutamate release from activated microglia requires the oxidative burst and lipid peroxidation. J Neurochem 101:1205–1213

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ben Sahra I, Regazzetti C, Robert G, Laurent K, Le Marchand-Brustel Y, Auberger P, Tanti JF, Giorgetti-Peraldi S, Bost F (2011) Metformin, independent of AMPK, induces mTOR inhibition and cell-cycle arrest through REDD1. Cancer Res 71:4366–4372

    Article  CAS  PubMed  Google Scholar 

  • Braidy N, Brew BJ, Inestrosa NC, Chung R, Sachdev P, Guillemin GJ (2014) Changes in cathepsin D and beclin-1 mRNA and protein expression by the excitotoxin quinolinic acid in human astrocytes and neurons. Metab Brain Dis 29:873–883

    Article  CAS  PubMed  Google Scholar 

  • Brugarolas J, Lei K, Hurley RL, Manning BD, Reiling JH, Hafen E, Witters LA, Ellisen LW, Kaelin WG Jr (2004) Regulation of mTOR function in response to hypoxia by REDD1 and the TSC1/TSC2 tumor suppressor complex. Genes Dev 18:2893–2904

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cheng YL, Park JS, Manzanero S, Choi Y, Baik SH, Okun E, Gelderblom M, Fann DY, Magnus T, Launikonis BS, Mattson MP, Sobey CG, Jo DG, Arumugam TV (2014) Evidence that collaboration between HIF-1alpha and Notch-1 promotes neuronal cell death in ischemic stroke. Neurobiol Dis 62:286–295

    Article  CAS  PubMed  Google Scholar 

  • Colin-Gonzalez AL, Orozco-Ibarra M, Chanez-Cardenas ME, Rangel-Lopez E, Santamaria A, Pedraza-Chaverri J, Barrera-Oviedo D, Maldonado PD (2013) Heme oxygenase-1 (HO-1) upregulation delays morphological and oxidative damage induced in an excitotoxic/pro-oxidant model in the rat striatum. Neuroscience 231:91–101

    Article  CAS  PubMed  Google Scholar 

  • Colin-Gonzalez AL, Luna-Lopez A, Konigsberg M, Ali SF, Pedraza-Chaverri J, Santamaria A (2014) Early modulation of the transcription factor Nrf2 in rodent striatal slices by quinolinic acid, a toxic metabolite of the kynurenine pathway. Neuroscience 260:130–139

    Article  CAS  PubMed  Google Scholar 

  • DeYoung MP, Horak P, Sofer A, Sgroi D, Ellisen LW (2008) Hypoxia regulates TSC1/2-mTOR signaling and tumor suppression through REDD1-mediated 14-3-3 shuttling. Genes Dev 22:239–251

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Du F, Wu XM, Gong Q, He X, Ke Y (2011) Hyperthermia conditioned astrocyte-cultured medium protects neurons from ischemic injury by the up-regulation of HIF-1 alpha and the increased anti-apoptotic ability. Eur J Pharmacol 666:19–25

    Article  CAS  PubMed  Google Scholar 

  • Duong TT, Antao S, Ellis NA, Myers SJ, Witting PK (2008) Supplementation with a synthetic polyphenol limits oxidative stress and enhances neuronal cell viability in response to hypoxia-re-oxygenation injury. Brain Res 1219:8–18

    Article  CAS  PubMed  Google Scholar 

  • Essa MM, Braidy N, Vijayan KR, Subash S, Guillemin GJ (2013) Excitotoxicity in the pathogenesis of autism. Neurotox Res 23:393–400

    Article  CAS  PubMed  Google Scholar 

  • Fan X, Heijnen CJ, van der Kooij MA, Groenendaal F, van Bel F (2009) The role and regulation of hypoxia-inducible factor-1alpha expression in brain development and neonatal hypoxic-ischemic brain injury. Brain Res Rev 62:99–108

    Article  CAS  PubMed  Google Scholar 

  • Ferrer CM, Lynch TP, Sodi VL, Falcone JN, Schwab LP, Peacock DL, Vocadlo DJ, Seagroves TN, Reginato MJ (2014) O-GlcNAcylation regulates cancer metabolism and survival stress signaling via regulation of the HIF-1 pathway. Mol Cell 54:820–831

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guillemin GJ, Brew BJ, Noonan CE, Takikawa O, Cullen KM (2005) Indoleamine 2,3 dioxygenase and quinolinic acid immunoreactivity in Alzheimer’s disease hippocampus. Neuropathol Appl Neurobiol 31:395–404

    Article  CAS  PubMed  Google Scholar 

  • Helton R, Cui J, Scheel JR, Ellison JA, Ames C, Gibson C, Blouw B, Ouyang L, Dragatsis I, Zeitlin S, Johnson RS, Lipton SA, Barlow C (2005) Brain-specific knock-out of hypoxia-inducible factor-1alpha reduces rather than increases hypoxic-ischemic damage. J Neurosci 25:4099–4107

    Article  CAS  PubMed  Google Scholar 

  • Horak P, Crawford AR, Vadysirisack DD, Nash ZM, DeYoung MP, Sgroi D, Ellisen LW (2010) Negative feedback control of HIF-1 through REDD1-regulated ROS suppresses tumorigenesis. Proc Natl Acad Sci U S A 107:4675–4680

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jeong JK, Park SY (2012) Transcriptional regulation of specific protein 1 (SP1) by hypoxia-inducible factor 1 alpha (HIF-1alpha) leads to PRNP expression and neuroprotection from toxic prion peptide. Biochem Biophys Res Commun 429:93–98

    Article  CAS  PubMed  Google Scholar 

  • Jiang H, Huang Y, Xu H, Sun Y, Han N, Li QF (2012) Hypoxia inducible factor-1alpha is involved in the neurodegeneration induced by isoflurane in the brain of neonatal rats. J Neurochem 120:453–460

    Article  CAS  PubMed  Google Scholar 

  • Lin C, Wu CJ, Wei IH, Tsai MH, Chang NW, Yang TT, Kuo YM (2013) Chronic treadmill running protects hippocampal neurons from hypobaric hypoxia-induced apoptosis in rats. Neuroscience 231:216–224

    Article  CAS  PubMed  Google Scholar 

  • Lopez-Hernandez B, Cena V, Posadas I (2015) The endoplasmic reticulum stress and the HIF-1 signalling pathways are involved in the neuronal damage caused by chemical hypoxia. Br J Pharmacol

  • Lugo-Huitron R, Ugalde Muniz P, Pineda B, Pedraza-Chaverri J, Rios C, Perez-de la Cruz V (2013) Quinolinic acid: an endogenous neurotoxin with multiple targets. Oxid Med Cell Longev 2013:104024

    Article  PubMed  PubMed Central  Google Scholar 

  • Maalouf M, Sullivan PG, Davis L, Kim DY, Rho JM (2007) Ketones inhibit mitochondrial production of reactive oxygen species production following glutamate excitotoxicity by increasing NADH oxidation. Neuroscience 145:256–264

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Malagelada C, Ryu EJ, Biswas SC, Jackson-Lewis V, Greene LA (2006) RTP801 is elevated in Parkinson brain substantia nigral neurons and mediates death in cellular models of Parkinson’s disease by a mechanism involving mammalian target of rapamycin inactivation. J Neurosci 26:9996–10005

    Article  CAS  PubMed  Google Scholar 

  • Malagelada C, Jin ZH, Jackson-Lewis V, Przedborski S, Greene LA (2010) Rapamycin protects against neuron death in in vitro and in vivo models of Parkinson’s disease. J Neurosci 30:1166–1175

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Martin-Flores N, Romani-Aumedes J, Rue L, Canal M, Sanders P, Straccia M, Allen ND, Alberch J, Canals JM, Perez-Navarro E, Malagelada C (2015) RTP801 is involved in mutant huntingtin-induced cell death. Mol Neurobiol

  • Mattson MP (2007) Calcium and neurodegeneration. Aging Cell 6:337–350

    Article  CAS  PubMed  Google Scholar 

  • Mehta A, Prabhakar M, Kumar P, Deshmukh R, Sharma PL (2013) Excitotoxicity: bridge to various triggers in neurodegenerative disorders. Eur J Pharmacol 698:6–18

    Article  CAS  PubMed  Google Scholar 

  • Meijer TW, Kaanders JH, Span PN, Bussink J (2012) Targeting hypoxia, HIF-1, and tumor glucose metabolism to improve radiotherapy efficacy. Clin Cancer Res 18:5585–5594

    Article  CAS  PubMed  Google Scholar 

  • Mishra J, Chaudhary T, Kumar A (2014) Rosiglitazone synergizes the neuroprotective effects of valproic acid against quinolinic acid-induced neurotoxicity in rats: targeting PPARgamma and HDAC pathways. Neurotox Res 26:130–151

    Article  CAS  PubMed  Google Scholar 

  • Nakai M, Qin ZH, Wang Y, Chase TN (1999) Free radical scavenger OPC-14117 attenuates quinolinic acid-induced NF-kappaB activation and apoptosis in rat striatum. Brain Res Mol Brain Res 64:59–68

    Article  CAS  PubMed  Google Scholar 

  • Noseda R, Belin S, Piguet F, Vaccari I, Scarlino S, Brambilla P, Martinelli Boneschi F, Feltri ML, Wrabetz L, Quattrini A, Feinstein E, Huganir RL, Bolino A (2013) DDIT4/REDD1/RTP801 is a novel negative regulator of Schwann cell myelination. J Neurosci 33:15295–15305

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pierozan P, Fernandes CG, Dutra MF, Pandolfo P, Ferreira F, de Lima BO, Porciuncula L, Wajner M, Pessoa-Pureur R (2014) Biochemical, histopathological and behavioral alterations caused by intrastriatal administration of quinolic acid to young rats. FEBS J 281:2061–2073

    Article  CAS  PubMed  Google Scholar 

  • Piret JP, Mottet D, Raes M, Michiels C (2002) Is HIF-1alpha a pro- or an anti-apoptotic protein? Biochem Pharmacol 64:889–892

    Article  CAS  PubMed  Google Scholar 

  • Rahmani M, Aust MM, Attkisson E, Williams DC Jr, Ferreira-Gonzalez A, Grant S (2013) Dual inhibition of Bcl-2 and Bcl-xL strikingly enhances PI3K inhibition-induced apoptosis in human myeloid leukemia cells through a GSK3- and Bim-dependent mechanism. Cancer Res 73:1340–1351

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rayner BS, Duong TT, Myers SJ, Witting PK (2006) Protective effect of a synthetic anti-oxidant on neuronal cell apoptosis resulting from experimental hypoxia re-oxygenation injury. J Neurochem 97:211–221

    Article  CAS  PubMed  Google Scholar 

  • Regazzetti C, Bost F, Le Marchand-Brustel Y, Tanti JF, Giorgetti-Peraldi S (2010) Insulin induces REDD1 expression through hypoxia-inducible factor 1 activation in adipocytes. J Biol Chem 285:5157–5164

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Reiling JH, Hafen E (2004) The hypoxia-induced paralogs Scylla and Charybdis inhibit growth by down-regulating S6K activity upstream of TSC in Drosophila. Genes Dev 18:2879–2892

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ryu JK, Choi HB, McLarnon JG (2005) Peripheral benzodiazepine receptor ligand PK11195 reduces microglial activation and neuronal death in quinolinic acid-injected rat striatum. Neurobiol Dis 20:550–561

    Article  CAS  PubMed  Google Scholar 

  • Schwarzer R, Tondera D, Arnold W, Giese K, Klippel A, Kaufmann J (2005) REDD1 integrates hypoxia-mediated survival signaling downstream of phosphatidylinositol 3-kinase. Oncogene 24:1138–1149

    Article  CAS  PubMed  Google Scholar 

  • Semenza GL (2000) HIF-1 and human disease: one highly involved factor. Genes Dev 14:1983–1991

    CAS  PubMed  Google Scholar 

  • Semenza GL (2010) HIF-1: upstream and downstream of cancer metabolism. Curr Opin Genet Dev 20:51–56

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Seo JS, Seol JW, Moon MH, Jeong JK, Lee YJ, Park SY (2010) Hypoxia protects neuronal cells from human prion protein fragment-induced apoptosis. J Neurochem 112:715–722

    Article  CAS  PubMed  Google Scholar 

  • Shoshani T, Faerman A, Mett I, Zelin E, Tenne T, Gorodin S, Moshel Y, Elbaz S, Budanov A, Chajut A, Kalinski H, Kamer I, Rozen A, Mor O, Keshet E, Leshkowitz D, Einat P, Skaliter R, Feinstein E (2002) Identification of a novel hypoxia-inducible factor 1-responsive gene, RTP801, involved in apoptosis. Mol Cell Biol 22:2283–2293

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Stoy N, Mackay GM, Forrest CM, Christofides J, Egerton M, Stone TW, Darlington LG (2005) Tryptophan metabolism and oxidative stress in patients with Huntington’s disease. J Neurochem 93:611–623

    Article  CAS  PubMed  Google Scholar 

  • Szydlowska K, Tymianski M (2010) Calcium, ischemia and excitotoxicity. Cell Calcium 47:122–129

    Article  CAS  PubMed  Google Scholar 

  • Wu XM, Qian ZM, Zhu L, Du F, Yung WH, Gong Q, Ke Y (2011) Neuroprotective effect of ligustilide against ischaemia-reperfusion injury via up-regulation of erythropoietin and down-regulation of RTP801. Br J Pharmacol 164:332–343

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yoshida T, Mett I, Bhunia AK, Bowman J, Perez M, Zhang L, Gandjeva A, Zhen L, Chukwueke U, Mao T, Richter A, Brown E, Ashush H, Notkin N, Gelfand A, Thimmulappa RK, Rangasamy T, Sussan T, Cosgrove G, Mouded M, Shapiro SD, Petrache I, Biswal S, Feinstein E, Tuder RM (2010) Rtp801, a suppressor of mTOR signaling, is an essential mediator of cigarette smoke-induced pulmonary injury and emphysema. Nat Med 16:767–773

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yuan G, Khan SA, Luo W, Nanduri J, Semenza GL, Prabhakar NR (2011) Hypoxia-inducible factor 1 mediates increased expression of NADPH oxidase-2 in response to intermittent hypoxia. J Cell Physiol 226:2925–2933

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zaman K, Ryu H, Hall D, O’Donovan K, Lin KI, Miller MP, Marquis JC, Baraban JM, Semenza GL, Ratan RR (1999) Protection from oxidative stress-induced apoptosis in cortical neuronal cultures by iron chelators is associated with enhanced DNA binding of hypoxia-inducible factor-1 and ATF-1/CREB and increased expression of glycolytic enzymes, p21(waf1/cip1), and erythropoietin. J Neurosci 19:9821–9830

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This research was supported by the fund of the National Natural Science Foundation of China (81300059 and 81400911), Natural Science Foundation of Jiangsu Province (bk20140573), China Postdoctoral Science Foundation (2014M560408) and Postdoctoral Science Foundation of Jiangsu Province (1402172C).

Author information

Authors and Affiliations

  1. Department of Pharmacology, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu, 212013, China

    Xiaojia Huang, Kaiyong Yang, Yi Zhang & Yongjin Li

  2. Department of Preventive Medicine, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu, 212013, China

    Qiang Wang

Authors
  1. Xiaojia Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kaiyong Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yi Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Qiang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yongjin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xiaojia Huang.

Ethics declarations

Conflict of interest

The authors have declared that no conflict of interest exists.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Huang, X., Yang, K., Zhang, Y. et al. Quinolinic acid induces cell apoptosis in PC12 cells through HIF-1-dependent RTP801 activation. Metab Brain Dis 31, 435–444 (2016). https://doi.org/10.1007/s11011-015-9782-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11011-015-9782-x

Keywords

Search

Navigation