HIF‑1α attenuates neuronal apoptosis by upregulating EPO expression following cerebral ischemia‑reperfusion injury in a rat MCAO model

Li,Jun; Tao,Tao; Xu,Jian; Liu,Zhi; Zou,Zhehua; Jin,Minglu

doi:10.3892/ijmm.2020.4480

HIF‑1α attenuates neuronal apoptosis by upregulating EPO expression following cerebral ischemia‑reperfusion injury in a rat MCAO model

Authors:
- Jun Li
- Tao Tao
- Jian Xu
- Zhi Liu
- Zhehua Zou
- Minglu Jin
View Affiliations

Affiliations: Department of Rehabilitation Medicine, Guizhou Provincial People's Hospital, Guiyang, Guizhou 550002, P.R. China, Department of Neurology, The Affiliated Hospital Guizhou Medical University, Guiyang, Guizhou 550001, P.R. China, Department of Pharmacy, The Affiliated Hospital Guizhou Medical University, Guiyang, Guizhou 550001, P.R. China, Department of General Practice, The First Hospital of Qinhuangdao, Qinhuangdao, Hebei 066000, P.R. China, Department of Neurology, Qijiang Hospital of The First Affiliated Hospital of Chongqing Medical University, Chongqing 404100, P.R. China
Published online on: January 28, 2020 https://doi.org/10.3892/ijmm.2020.4480
Pages: 1027-1036
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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‑inducible factor‑1α (HIF‑1α) is a key transcriptional factor in response to hypoxia and is involved in ischemic stroke. In the present study, the potential for HIF‑1α to inhibit neuronal apoptosis through upregulating erythropoietin (EPO) was investigated in a transient middle cerebral artery occlusion (tMCAO) rat stroke model. For this purpose, a recombinant adenovirus expressing HIF‑1α was engineered (Ad‑HIF‑1α). Control adenovirus (Ad group), Ad‑HIF‑1α (Ad‑HIF‑1α group) or Ad‑HIF‑1α in addition to erythropoietin mimetic peptide‑9 (EMP9), an EPO‑receptor (‑R) antagonist (Ad‑HIF‑1α+EMP9 group), were used for an intracranial injection into rat ischemic penumbra 1 h following MCAO. All rats demonstrated functional improvement following tMCAO, while the improvement rate was faster in rats treated by Ad‑HIF‑1α compared with all other groups. The EPO‑R inhibitor partially reversed the benefits of Ad‑HIF‑1α. Apoptosis induced by tMCAO was significantly inhibited by Ad‑HIF‑1α (P<0.05). The expression of HIF‑1α, evaluated by immunohistochemistry either in neurons or astrocytes, was upregulated by Ad‑HIF‑1α. Both EPO mRNA and protein expression were increased by Ad‑HIF‑1α, however, there was no significant change of EPO‑R either on an mRNA level or protein level. Furthermore, EMP9 did not change the EPO expression which was upregulated by Ad‑HIF‑1α. Activated caspase 3 in neurons was suppressed by Ad‑HIF‑1α. Activated caspase 3 downregulated by HIF‑1α was partially blocked by EMP9. Altogether, the present data demonstrated that HIF‑1α attenuates neuronal apoptosis partially through upregulating EPO following cerebral ischemia in rat. Thus, upregulating HIF‑1α subsequent to a stroke may be a potential treatment for ischemic stroke.

View Figures

View References

1	Ng M, Fleming T, Robinson M, Thomson B, Graetz N, Margono C, Mullany EC, Biryukov S, Abbafati C, Abera SF, et al: Global, regional, and national prevalence of overweight and obesity in children and adults during 1980-2013: A systematic analysis for the Global Burden of Disease Study 2013. Lancet. 384:766–781. 2014. View Article : Google Scholar : PubMed/NCBI
2	GBD 2016 Causes of Death Collaborators: Global, regional, and national age-sex specific mortality for 264 causes of death, 1980-2016: A systematic analysis for the Global Burden of Disease Study 2016. Lancet. 390:1151–210. 2017. View Article : Google Scholar : PubMed/NCBI
3	Yang G, Wang Y, Zeng Y, Gao GF, Liang X, Zhou M, Wan X, Yu S, Jiang Y, Naghavi M, et al: Rapid health transition in China, 1990-2010: Findings from the global burden of disease study 2010. Lancet. 381:1987–2015. 2013. View Article : Google Scholar : PubMed/NCBI
4	Liu L, Wang D, Wong KS and Wang Y: Stroke and stroke care in China: Huge burden, significant workload, and a national priority. Stroke. 42:3651–3654. 2011. View Article : Google Scholar : PubMed/NCBI
5	Wang W, Jiang B, Sun H, Ru X, Sun D, Wang L, Wang L, Jiang Y, Li Y, Wang Y, et al: Prevalence, incidence, and mortality of stroke in China: Results from a nationwide population-based survey of 480 687 adults. Circulation. 135:759–771. 2017. View Article : Google Scholar : PubMed/NCBI
6	Wang YJ, Zhang SM, Zhang L, Wang CX, Dong Q, Gao S, Huang RX, Huang YN, Lv CZ, Liu M, et al: Chinese guidelines for the secondary prevention of ischemic stroke and transient ischemic attack 2010. CNS Neurosci Ther. 18:93–101. 2012. View Article : Google Scholar : PubMed/NCBI
7	Hacke W, Kaste M, Bluhmki E, Brozman M, Davalos A, Guidetti D, Larrue V, Lees KR, Medeghri Z, Machnig T, et al: Thrombolysis with alteplase 3 to 4.5 hours after acute ischemic stroke. N Engl J Med. 359:1317–1329. 2008. View Article : Google Scholar : PubMed/NCBI
8	Xydas T, Georgantopoulos C, Bethanis D and Sarris M: Thrombolysis for acute ischemic stroke: A new paradigm. Hosp Chron. 7:77–80. 2012.
9	National 'Ninth Five-Year Plan' Research Group; Chen Q: Intravenous thrombolysis with urokinase for acute cerebral infarctions. Chin J Neurol. 35:210–213. 2002.In Chinese.
10	Semenza GL: Targeting HIF-1 for cancer therapy. Nat Rev Cancer. 3:721–732. 2003. View Article : Google Scholar : PubMed/NCBI
11	Semenza GL: Hypoxia-inducible factor 1: Oxygen homeostasis and disease pathophysiology. Trends Mol Med. 7:345–350. 2001. View Article : Google Scholar : PubMed/NCBI
12	Ivan M, Kondo K, Yang H, Kim W, Valiando J, Ohh M, Salic A, Asara JM, Lane WS and Kaelin WG Jr: HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: Implications for O2 sensing. Science. 292:464–468. 2001. View Article : Google Scholar : PubMed/NCBI
13	Abe H, Semba H and Takeda N: The roles of hypoxia signaling in the pathogenesis of cardiovascular diseases. J Atheroscler Thromb. 24:884–894. 2017. View Article : Google Scholar : PubMed/NCBI
14	Feng Y and Bhatt AJ: Corticosteroid responses following hypoxic preconditioning provide neuroprotection against subsequent hypoxic-ischemic brain injury in the newborn rats. Int J Dev Neurosci. 44:6–13. 2015. View Article : Google Scholar : PubMed/NCBI
15	Chen J, Yang Y, Shen L, Ding W, Chen X, Wu E, Cai K and Wang G: Hypoxic preconditioning augments the therapeutic efficacy of bone marrow stromal cells in a rat ischemic stroke model. Cell Mol Neurobiol. 37:1115–1129. 2017. View Article : Google Scholar
16	Jacobs K, Shoemaker C, Rudersdorf R, Neill SD, Kaufman RJ, Mufson A, Seehra J, Jones SS, Hewick R and Fritsch EF: Isolation and characterization of genomic and cDNA clones of human erythropoietin. Nature. 313:806–810. 1985. View Article : Google Scholar : PubMed/NCBI
17	Chikuma M, Masuda S, Kobayashi T, Nagao M and Sasaki R: Tissue-specific regulation of erythropoietin production in the murine kidney, brain, and uterus. Am J Physiol Endocrinol Metab. 279:E1242–E1248. 2000. View Article : Google Scholar : PubMed/NCBI
18	Yu X, Shacka JJ, Eells JB, Suarez-Quian C, Przygodzki RM, Beleslin-Cokic B, Lin CS, Nikodem VM, Hempstead B, Flanders KC, et al: Erythropoietin receptor signalling is required for normal brain development. Development. 129:505–516. 2002.PubMed/NCBI
19	Iwai M, Stetler RA, Xing J, Hu X, Gao Y, Zhang W, Chen J and Cao G: Enhanced oligodendrogenesis and recovery of neurological function by erythropoietin after neonatal hypoxic/ischemic brain injury. Stroke. 41:1032–1037. 2010. View Article : Google Scholar : PubMed/NCBI
20	Wu SH, Lu IC, Lee SS, Kwan AL, Chai CY and Huang SH: Erythropoietin attenuates motor neuron programmed cell death in a burn animal model. PLoS One. 13:e01900392018. View Article : Google Scholar : PubMed/NCBI
21	Wang P, Lu Y, Han D, Wang P, Ren L, Bi J and Liang J: Neuroprotection by nicotinamide mononucleotide adenylyltransferase 1 with involvement of autophagy in an aged rat model of transient cerebral ischemia and reperfusion. Brain Res. 1723:1463912019. View Article : Google Scholar : PubMed/NCBI
22	Saadoun S, Waters P, Bell BA, Vincent A, Verkman AS and Papadopoulos MC: Intra-cerebral injection of neuromyelitis optica immunoglobulin G and human complement produces neuromyelitis optica lesions in mice. Brain. 133:349–361. 2010. View Article : Google Scholar : PubMed/NCBI
23	Chen T, Yu Y, Tang LJ, Kong L, Zhang CH, Chu HY, Yin LW and Ma HY: Neural stem cells over-expressing brain-derived neurotrophic factor promote neuronal survival and cytoskeletal protein expression in traumatic brain injury sites. Neural Regen Res. 12:433–439. 2017. View Article : Google Scholar : PubMed/NCBI
24	Wu W, Chen X, Hu C, Li J, Yu Z and Cai W: Transplantation of neural stem cells expressing hypoxia-inducible factor-1alpha (HIF-1alpha) improves behavioral recovery in a rat stroke model. J Clin Neurosci. 17:92–95. 2010. View Article : Google Scholar
25	Yang ML, Tao T, Xu J, Liu Z and Xu D: Antiapoptotic effect of gene therapy with recombinant adenovirus vector containing hypoxia-inducible factor-1α after cerebral ischemia and reperfu-sion in rats. Chin Med J (Engl). 130:1700–1706. 2017. View Article : Google Scholar
26	Lin YW and Hsieh CL: Electroacupuncture at Baihui acupoint (GV20) reverses behavior deficit and long-term potentiation through N-methyl-d-aspartate and transient receptor potential vanilloid subtype 1 receptors in middle cerebral artery occlusion rats. J Integr Neurosci. 9:269–282. 2010. View Article : Google Scholar : PubMed/NCBI
27	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
28	Snigdha S, Smith ED, Prieto GA and Cotman CW: Caspase-3 activation as a bifurcation point between plasticity and cell death. Neurosci Bull. 28:14–24. 2012. View Article : Google Scholar : PubMed/NCBI
29	Li Y, Huang L, Ma Q, Concepcion KR, Song MA, Zhang P, Fu Y, Xiao D and Zhang L: Repression of the glucocorticoid receptor aggravates acute ischemic brain injuries in adult mice. Int J Mol Sci. 19:E24282018. View Article : Google Scholar : PubMed/NCBI
30	Paciaroni M, Caso V and Agnelli G: The concept of ischemic penumbra in acute stroke and therapeutic opportunities. Eur Neurol. 61:321–330. 2009. View Article : Google Scholar : PubMed/NCBI
31	Rosso C and Samson Y: The ischemic penumbra: The location rather than the volume of recovery determines outcome. Curr Opin Neurol. 27:35–41. 2014. View Article : Google Scholar
32	Ran YC, Zhu M, Li SJ, Zhang ZX, Wang X, Zhang Y and Cheng JL: Related research and recent progress of ischemic penumbra. World Neurosurg. 116:5–13. 2018. View Article : Google Scholar : PubMed/NCBI
33	Murphy TH and Corbett D: Plasticity during stroke recovery: From synapse to behaviour. Nat Rev Neurosci. 10:861–872. 2009. View Article : Google Scholar : PubMed/NCBI
34	Reischl S, Li L, Walkinshaw G, Flippin LA, Marti HH and Kunze R: Inhibition of HIF prolyl-4-hydroxylases by FG-4497 reduces brain tissue injury and edema formation during ischemic stroke. PLoS One. 9:e847672014. View Article : Google Scholar : PubMed/NCBI
35	Ogle ME, Gu X, Espinera AR and Wei L: Inhibition of prolyl hydroxylases by dimethyloxaloylglycine after stroke reduces ischemic brain injury and requires hypoxia inducible factor-1α. Neurobiol Dis. 45:733–742. 2012. View Article : Google Scholar
36	Li C, Zhang B, Zhu Y, Li Y, Liu P, Gao B, Tian S, Du L and Bai Y: Post-stroke constraint-induced movement therapy increases functional recovery, angiogenesis, and neurogenesis with enhanced expression of HIF-1α and VEGF. Curr Neurovasc Res. 14:368–377. 2017. View Article : Google Scholar
37	Wang H, Niu F, Fan W, Shi J, Zhang J and Li B: Modulating effects of preconditioning exercise in the expression of ET-1 and BNP via HIF-1α in ischemically injured brain. Metab Brain Dis. 34:1299–1311. 2019. View Article : Google Scholar : PubMed/NCBI
38	Gao Y, Yin H, Zhang Y, Dong Y, Yang F, Wu X and Liu H: Dexmedetomidine protects hippocampal neurons against hypoxia/reoxygenation-induced apoptosis through activation HIF-1α/p53 signaling. Life Sci. 232:1166112019. View Article : Google Scholar
39	Baranova O, Miranda LF, Pichiule P, Dragatsis I, Johnson RS and Chavez JC: 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. 2007. View Article : Google Scholar : PubMed/NCBI
40	Barteczek P, Li L, Ernst AS, Böhler LI, Marti HH and Kunze R: Neuronal HIF-1α and HIF-2α deficiency improves neuronal survival and sensorimotor function in the early acute phase after ischemic stroke. J Cereb Blood Flow Metab. 37:291–306. 2017. View Article : Google Scholar
41	Helton R, Cui J, Scheel JR, Ellison JA, Ames C, Gibson C, Blouw B, Ouyang L, Dragatsis I, Zeitlin S, et al: Brain-specific knock-out of hypoxia-inducible factor-1 alpha reduces rather than increases hypoxic-ischemic damage. J Neurosci. 25:4099–4107. 2005. View Article : Google Scholar : PubMed/NCBI
42	Yan J, Zhou B, Taheri S and Shi H: Differential effects of HIF-1 inhibition by YC-1 on the overall outcome and blood-brain barrier damage in a rat model of ischemic stroke. PLoS One. 6:e277982011. View Article : Google Scholar : PubMed/NCBI
43	McIlwain DR, Berger T and Mak TW: Caspase functions in cell death and disease. Cold Spring Harb Perspect Boil. 5:a0086562013.
44	O'Donovan N, Crown J, Stunell H, Hill AD, McDermott E, O'Higgins N and Duffy MJ: Caspase 3 in breast cancer. Clin Cancer Res. 9:738–742. 2003.PubMed/NCBI
45	Digicaylioglu M and Lipton SA: Erythropoietin-mediated neuroprotection involves cross-talk between Jak2 and NF-kappaB signalling cascades. Nature. 412:641–647. 2001. View Article : Google Scholar : PubMed/NCBI
46	Bouscary D, Pene F, Claessens YE, Muller O, Chrétien S, Fontenay-Roupie M, Gisselbrecht S, Mayeux P and Lacombe C: Critical role for PI 3-kinase in the control of erythropoietininduced erythroid progenitor proliferation. Blood. 101:3436–3443. 2003. View Article : Google Scholar
47	Haq R, Halupa A, Beattie BK, Mason JM, Zanke BW and Barber DL: Regulation of erythropoietin-induced STAT serine phosphorylation by distinct mitogen-activated protein kinases. J Biol Chem. 277:17359–17366. 2002. View Article : Google Scholar : PubMed/NCBI
48	Zhi-Kun S, Hong-Qi Y, Zhi-Quan W, Jing P, Zhen H and Sheng-Di C: Erythropoietin prevents PC12 cells from beta-amyloid-induced apoptosis via PI3K⁄Akt pathway. Transl Neurodegener. 1:72012. View Article : Google Scholar
49	Fan X, Heijnen CJ, van der KOOIJ MA, Groenendaal F and van Bel F: Beneficial effect of erythropoietin on sensorimotor function and white matter after hypoxia-ischemia in neonatal mice. Pediatr Res. 69:56–61. 2011. View Article : Google Scholar
50	Wang Y, Zhang H, Liu Y, Li P, Cao Z and Cao Y: Erythropoietin (EPO) protects against high glucose-induced apoptosis in retinal ganglional cells. Cell Biochem Biophys. 71:749–755. 2015. View Article : Google Scholar
51	Cheng L, Yu H, Yan N, Lai K and Xiang M: Hypoxia-inducible factor-1α target genes contribute to retinal neuroprotection. Front Cell Neurosci. 11:202017. View Article : Google Scholar
52	Wang L, Zhao RP, Song XY and Wu WF: Targeting ERβ in macrophage reduces crown-like structures in adipose tissue by inhibiting osteopontin and HIF-1α. Sci Rep. 9:157622019. View Article : Google Scholar
53	Lee SH, Jeong D, Han YS and Baek MJ: Pivotal role of vascular endothelial growth factor pathway in tumor angiogenesis. Ann Surg Treat Res. 89:1–8. 2015. View Article : Google Scholar : PubMed/NCBI
54	Charriaut-Marlangue C, Margaill I, Represa A, Popovici T, Plotkine M and Ben-Ari Y: Apoptosis and necrosis after reversible focal ischemia: An in situ DNA fragmentation analysis. J Cereb Blood Flow Metab. 16:186–194. 1996. View Article : Google Scholar : PubMed/NCBI