Time‑dependent homeostasis between glucose uptake and consumption in astrocytes exposed to CoCl2 treatment

Wang,Peng; Li,Ling; Zhang,Zhenxiang; Kan,Quancheng; Chen,Suyan; Gao,Feng

doi:10.3892/mmr.2016.4873

Time‑dependent homeostasis between glucose uptake and consumption in astrocytes exposed to CoCl2 treatment

Authors:
- Peng Wang
- Ling Li
- Zhenxiang Zhang
- Quancheng Kan
- Suyan Chen
- Feng Gao
View Affiliations

Affiliations: Department of Basic Medicine, Nursing College, Zhengzhou University, Zhengzhou, Henan 450052, P.R. China, Department of Palliative Care and Hospice Care, The Ninth People's Hospital of Zhengzhou, Zhengzhou, Henan 450053, P.R. China, Department of Infectious Disease, Clinical Pharmacology Base, First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China, Department of Neuroimmunology, Henan Academy of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou, Henan 450052, P.R. China
Published online on: February 5, 2016 https://doi.org/10.3892/mmr.2016.4873
Pages: 2909-2917

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

Hypoxia has been implicated in the pathology of the central nervous system during stroke. It also has a significant effect on the regulation of glucose transporters (GLUTs), and homeostasis between glucose uptake and consumption. CoCl2 is a hypoxia‑mimetic agent, and thus stabilizes the hypoxia‑inducible factor 1α (HIF‑1α) subunit and regulates GLUT genes. GLUT‑1 and GLUT‑3 are the most common isoforms of the GLUT family present in the brain, with the former primarily expressed in astrocytes and the latter in neurons under physiological conditions. However, it remains controversial whether GLUT‑3 is expressed in astrocytes. Additionally, it is unclear whether the regulation of GLUT‑1 and GLUT‑3, and glucose homeostasis, are affected by CoCl2 treatment in a time‑dependent manner. In the present study, mRNA and protein levels of GLUT‑1, GLUT‑3 and HIF‑1α in astrocytes were examined by reverse transcription‑quantitative polymerase chain reaction and western blot analysis, respectively. The intracellular glucose concentration, glycogen storage, ATP content, pyruvate concentration, lactate dehydrogenase (LDH) release activity and cell viability in astrocytes were also investigated. The observations of the current study confirmed that both protein and mRNA levels of GLUT‑1 and GLUT‑3 were elevated in a time‑dependent manner induced by CoCl2 treatment, followed by accumulation of HIF‑1α. Furthermore, in the early period of CoCl2 treatment (≤8 h at 100 µM), LDH release, ATP content, glycogen storage and cell viability remained unchanged, whereas intracellular pyruvate concentration increased and glucose concentration was reduced. However, in the later period of CoCl2 treatment (>8 h at 100 µM), LDH release and intracellular pyruvate concentration increased, while intracellular glucose concentration, ATP content and glycogen storage were reduced. This may be due to disruption of homeostasis and reduced cell viability. In conclusion, alteration in the expression levels of GLUT‑1 and GLUT‑3, and the homeostasis between glucose uptake and consumption were affected by CoCl2 treatment, in a time‑dependent manner, and may result in reduced energy production and cell viability in astrocytes.

View Figures

View References

1	Cunningham LA, Candelario K and Li L: Roles for HIF-1α in neural stem cell function and the regenerative response to stroke. Behav Brain Res. 227:410–417. 2012. View Article : Google Scholar
2	Wang C, Wang Z, Zhang X, Zhang X, Dong L, Xing Y, Li Y, Liu Z, Chen L, Qiao H, et al: Protection by silibinin against experimental ischemic stroke: up-regulated pAkt, pmTOR, HIF-1α and Bcl-2, down-regulated Bax, NF-κB expression. Neurosci Lett. 529:45–50. 2012. View Article : Google Scholar : PubMed/NCBI
3	Zemke D, Smith JL, Reeves MJ and Majid A: Ischemia and ischemic tolerance in the brain: An overview. Neurotoxicology. 25:895–904. 2004. View Article : Google Scholar : PubMed/NCBI
4	Ratan RR, Siddiq A, Smirnova N, Karpisheva K, Haskew-Layton R, McConoughey S, Langley B, Estevez A, Huerta PT, Volpe B, et al: Harnessing hypoxic adaptation to prevent, treat, and repair stroke. J Mol Med Berl. 85:1331–1338. 2007. View Article : Google Scholar : PubMed/NCBI
5	Vemula S, Roder KE, Yang T, Bhat GJ, Thekkumkara TJ and Abbruscato TJ: A functional role for sodium-dependent glucose transport across the blood-brain barrier during oxygen glucose deprivation. J Pharmacol Exp Ther. 328:487–495. 2009. View Article : Google Scholar :
6	Agrawal M, Kumar V, Kashyap MP, Khanna VK, Randhawa GS and Pant AB: Ischemic insult induced apoptotic changes in PC12 cells: protection by trans resveratrol. Eur J Pharmacol. 666:5–11. 2011. View Article : Google Scholar : PubMed/NCBI
7	Reagan LP, Magariños AM, Lucas LR, van Bueren A, McCall AL and McEwen BS: Regulation of GLUT-3 glucose transporter in the hippocampus of diabetic rats subjected to stress. Am J Physiol. 276:E879–E886. 1999.PubMed/NCBI
8	Iwabuchi S and Kawahara K: Inducible astrocytic glucose transporter-3 contributes to the enhanced storage of intracellular glycogen during reperfusion after ischemia. Neurochem Int. 59:319–325. 2011. View Article : Google Scholar : PubMed/NCBI
9	Badawi Y, Ramamoorthy P and Shi H: Hypoxia-inducible factor 1 protects hypoxic astrocytes against glutamate toxicity. ASN Neuro. 4:231–241. 2012. View Article : Google Scholar : PubMed/NCBI
10	Valle-Casuso JC, González-Sánchez A, Medina JM and Tabernero A: HIF-1 and c-Src mediate increased glucose uptake induced by endothelin-1 and connexin43 in astrocytes. PLoS One. 7:e324482012. View Article : Google Scholar : PubMed/NCBI
11	Pereira KM, Chaves FN, Viana TS, Carvalho FS, Costa FW, Alves AP and Sousa FB: Oxygen metabolism in oral cancer: HIF and GLUT-s. Oncol Lett. 6:311–316. 2013.Review. PubMed/NCBI
12	Seagroves TN, Ryan HE, Lu H, Wouters BG, Knapp M, Thibault P, Laderoute K and Johnson RS: Transcription factor HIF-1 is a necessary mediator of the pasteur effect in mammalian cells. Mol Cell Biol. 21:3436–3444. 2001. View Article : Google Scholar : PubMed/NCBI
13	Kinni H, Guo M, Ding JY, Konakondla S, Dornbos D III, Tran R, Guthikonda M and Ding Y: Cerebral metabolism after forced or voluntary physical exercise. Brain Res. 1388:48–55. 2011. View Article : Google Scholar : PubMed/NCBI
14	Brix B, Mesters JR, Pellerin L and Jöhren O: Endothelial cell-derived nitric oxide enhances aerobic glycolysis in astrocytes via HIF-1α-mediated target gene activation. J Neurosci. 32:9727–9735. 2012. View Article : Google Scholar : PubMed/NCBI
15	Rouach N, Koulakoff A, Abudara V, Willecke K and Giaume C: Astroglial metabolic networks sustain hippocampal synaptic transmission. Science. 322:1551–1555. 2008. View Article : Google Scholar : PubMed/NCBI
16	Duelli R and Kuschinsky W: Brain glucose transporters: Relationship to local energy demand. News Physiol Sci. 16:71–76. 2001.PubMed/NCBI
17	Nijland PG, Michailidou I, Witte ME, Mizee MR, van der Pol SM, van Het Hof B, Reijerkerk A, Pellerin L, van der Valk P, de Vries HE and van Horssen J: Cellular distribution of glucose and monocarboxylate transporters in human brain white matter and multiple sclerosis lesions. Glia. 62:1125–1141. 2014. View Article : Google Scholar : PubMed/NCBI
18	Cidad P, Garcia-Nogales P, Almeida A and Bolaños JP: Expression of glucose transporter GLUT-3 by endotoxin in cultured rat astrocytes: the role of nitric oxide. J Neurochem. 79:17–24. 2001. View Article : Google Scholar : PubMed/NCBI
19	Maher F, Davies-Hill TM and Simpson IA: Substrate specificity and kinetic parameters of GLUT-3 in rat cerebellar granule neurons. Biochem J. 315:827–831. 1996. View Article : Google Scholar
20	Efrati S, Fishlev G, Bechor Y, Volkov O, Bergan J, Kliakhandler K, Kamiager I, Gal N, Friedman M, Ben-Jacob E and Golan H: Hyperbaric oxygen induces late neuroplasticity in post stroke patients - randomized, prospective trial. PLoS One. 8:e537162013. View Article : Google Scholar :
21	Saxena S, Shukla D and Bansal A: Augmentation of aerobic respiration and mitochondrial biogenesis in skeletal muscle by hypoxia preconditioning with cobalt chloride. Toxicol Appl Pharmacol. 264:324–334. 2012. View Article : Google Scholar : PubMed/NCBI
22	Björklund O, Shang M, Tonazzini I, Daré E and Fredholm BB: Adenosine A1 and A3 receptors protect astrocytes from hypoxic damage. Eur J Pharmacol. 596:6–13. 2008. View Article : Google Scholar : PubMed/NCBI
23	McCarthy KD and de Vellis J: Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol. 85:890–902. 1980. View Article : Google Scholar : PubMed/NCBI
24	Karovic O, Tonazzini I, Rebola N, Edström E, Lövdahl C, Fredholm BB and Daré E: Toxic effects of cobalt in primary cultures of mouse Astrocytes Similarities with hypoxia and role of HIF-1a. Biochem Pharmacol. 73:694–708. 2007. View Article : Google Scholar
25	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
26	Schubert D: Glucose metabolism and Alzheimer's disease. Ageing Res Rev. 4:240–257. 2005. View Article : Google Scholar : PubMed/NCBI
27	Jones NM and Bergeron M: Hypoxic preconditioning induces changes in HIF-1 target genes in neonatal rat brain. J Cereb Blood Flow Metab. 21:1105–1114. 2001. View Article : Google Scholar : PubMed/NCBI
28	Wang KR, Jiang T, Wu TT, Zhou SH, Yao HT, Wang QY and Lu ZJ: Expression of hypoxia-related markers in inflammatory myofibroblastic tumors of the head and neck. World J Surg Oncol. 11:2942013. View Article : Google Scholar : PubMed/NCBI
29	Papa Pde C, Sousa LM, Silva Rdos S, de Fátima LA, da Fonseca VU, do Amaral VC, Hoffmann B, Alves-Wagner AB, Machado UF and Kowalewski MP: Glucose transporter 1 expression accompanies hypoxia sensing in the cyclic canine corpus luteum. Reproduction. 147:81–89. 2014. View Article : Google Scholar
30	Yamada T, Uchida M, Kwang-Lee K, Kitamura N, Yoshimura T, Sasabe E and Yamamoto T: Correlation of metabolism/hypoxia markers and fluorodeoxyglucose uptake in oral squamous cell carcinomas. Oral Surg Oral Med Oral Pathol Oral Radiol. 113:464–471. 2012. View Article : Google Scholar : PubMed/NCBI
31	Xu O, Li X, Qu Y, Liu S, An J, Wang M, Sun Q, Zhang W, Lu X, Pi L, et al: Regulation of glucose transporter protein-1 and vascular endothelial growth factor by hypoxia inducible factor 1α under hypoxic conditions in Hep-2 human cells. Mol Med Rep. 6:1418–1422. 2012.PubMed/NCBI
32	Dornbos D III, Zwagerman N, Guo M, Ding JY, Peng C, Esmail F, Sikharam C, Geng X, Guthikonda M and Ding Y: Preischemic exercise reduces brain damage by ameliorating metabolic disorder in ischemia/reperfusion injury. J Neurosci Res. 91:818–827. 2013. View Article : Google Scholar : PubMed/NCBI
33	MacVicar BA: Morphological differentiation of cultured astrocytes is blocked by cadmium or cobalt. Brain Res. 420:175–177. 1987. View Article : Google Scholar : PubMed/NCBI
34	Eckert AW, Lautner MH, Schütze A, Taubert H, Schubert J and Bilkenroth U: Coexpression of hypoxia-inducible factor-1α and glucose transporter-1 is associated with poor prognosis in oral squamous cell carcinoma patients. Histopathology. 58:1136–1147. 2011. View Article : Google Scholar : PubMed/NCBI