HSP90β regulates EAAT2 expression and participates in ischemia‑reperfusion injury in rats

Liang,Tao; Hu,Xiaolin; Zeng,Ling; Zhou,Zhiwei; Zhang,Jun; Xu,Zucai; Zeng,Junwei; Xu,Ping

doi:10.3892/mmr.2023.13128

HSP90β regulates EAAT2 expression and participates in ischemia‑reperfusion injury in rats

Authors:
- Tao Liang
- Xiaolin Hu
- Ling Zeng
- Zhiwei Zhou
- Jun Zhang
- Zucai Xu
- Junwei Zeng
- Ping Xu
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Affiliations: Soochow University Medical College, Soochow University, Suzhou, Jiangsu 215031, P.R. China, Department of Neurology, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou 563000, P.R. China, Department of Physiology, Zunyi Medical University, Zunyi, Guizhou 564699, P.R. China
Published online on: November 15, 2023 https://doi.org/10.3892/mmr.2023.13128
Article Number: 5
Copyright: © Liang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cerebrovascular diseases (CVDs) have become a global public health problem and ischemia‑reperfusion injury, the major cause of neurological impairment exacerbation, is closely related to excitotoxicity. The present study aimed to investigate the effects of changes in heat shock protein (HSP)90β expression and verify whether HSP90β regulates EAAT2 expression in a cerebral ischemia‑reperfusion injury model. Healthy adult Sprague‑Dawley (SD) male rats were used to establish a control group, sham‑operated group, middle cerebral artery occlusion (MCAO) group, empty virus group and lentivirus group. A model of cerebral ischemia‑reperfusion was established using the MCAO method. Lentivirus construction and injection were used to interfere with the expression of HSP90β. The modified neurological severity score was used to assess neurological deficits. Triphenyltetrazolium chloride staining was used to detect infarct areas. Immunofluorescence was used to detect HSP90β expression localization and the expression levels of HSP90β and EAAT2 were determined using western blotting and reverse transcription‑quantitative PCR. An MCAO model was successfully established and it was found that HSP90β, but not HSP90α, was upregulated after MCAO. HSP90β expression coincided with astrocyte markers in the ischemic penumbra area, while no expression was observed in microglia. Inhibition of HSP90β expression improved neurological deficits and alleviated brain injury by increasing EAAT2 expression. These results suggested that HSP90β is involved in the process of cerebral ischemia‑reperfusion injury in rats and that inhibition of HSP90β expression increases EAAT2 levels, conferring a neuroprotective effect in MCAO model rats.

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1	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
2	Wang D, Liu J, Liu M, Lu C, Brainin M and Zhang J: Patterns of stroke between university hospitals and nonuniversity hospitals in Mainland China: Prospective multicenter hospital-based registry study. World Neurosurg. 98:258–265. 2017. View Article : Google Scholar : PubMed/NCBI
3	Yuan Y, Tian Y, Jiang H, Cai LY, Song J, Peng R and Zhang XM: Mechanism of PGC-1α-mediated mitochondrial biogenesis in cerebral ischemia-reperfusion injury. Front Mol Neurosci. 16:12249642023. View Article : Google Scholar : PubMed/NCBI
4	Uzdensky AB: Apoptosis regulation in the penumbra after ischemic stroke: Expression of pro- and antiapoptotic proteins. Apoptosis. 24:687–702. 2019. View Article : Google Scholar : PubMed/NCBI
5	Chao XD, Fei F and Fei Z: The role of excitatory amino acid transporters in cerebral ischemia. Neurochem Res. 35:1224–1230. 2010. View Article : Google Scholar : PubMed/NCBI
6	Chi OZ, Hunter C, Liu X and Weiss HR: Effects of exogenous excitatory amino acid neurotransmitters on blood-brain barrier disruption in focal cerebral ischemia. Neurochem Res. 34:1249–1254. 2009. View Article : Google Scholar : PubMed/NCBI
7	Takahashi K, Foster JB and Lin CL: Glutamate transporter EAAT2: Regulation, function, and potential as a therapeutic target for neurological and psychiatric disease. Cell Mol Life Sci. 72:3489–3506. 2015. View Article : Google Scholar : PubMed/NCBI
8	Sankhala KK, Mita MM, Mita AC and Takimoto CH: Heat shock proteins: A potential anticancer target. Curr Drug Targets. 12:2001–2008. 2011. View Article : Google Scholar : PubMed/NCBI
9	Whitesell L and Lindquist SL: HSP90 and the chaperoning of cancer. Nat Rev Cancer. 5:761–772. 2005. View Article : Google Scholar : PubMed/NCBI
10	Grad I, Cederroth CR, Walicki J, Grey C, Barluenga S, Winssinger N, De Massy B, Nef S and Picard D: The molecular chaperone Hsp90α is required for meiotic progression of spermatocytes beyond pachytene in the mouse. PLoS One. 5:e157702010. View Article : Google Scholar : PubMed/NCBI
11	Zhang R, Liu C, Ji Y, Teng L and Guo Y: Neuregulin-1β plays a neuroprotective role by inhibiting the Cdk5 signaling pathway after cerebral ischemia-reperfusion injury in rats. J Mol Neurosci. 66:261–272. 2018. View Article : Google Scholar : PubMed/NCBI
12	Longa EZ, Weinstein PR, Carlson S and Cummins R: Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke. 20:84–91. 1989. View Article : Google Scholar : PubMed/NCBI
13	Yu X, Luo Y, Yang L, Chen P and Duan X: P-hydroxybenzyl alcohol ameliorates neuronal cerebral ischemia-reperfusion injury by activating mitochondrial autophagy through SIRT1. Mol Med Rep. 27:682023. View Article : Google Scholar : PubMed/NCBI
14	Zhou M, Wang H, Zeng X, Yin P, Zhu J, Chen W, Li X, Wang L, Wang L, Liu Y, et al: Mortality, morbidity, and risk factors in China and its provinces, 1990–2017: A systematic analysis for the global burden of disease study 2017. Lancet. 394:1145–1158. 2019. View Article : Google Scholar : PubMed/NCBI
15	GBD 2017 Causes of Death Collaborators, . Global, regional, and national age-sex-specific mortality for 282 causes of death in 195 countries and territories, 1980–2017: A systematic analysis for the global burden of disease study 2017. Lancet. 392:1736–1788. 2018. View Article : Google Scholar : PubMed/NCBI
16	Bradley E, Zhao X, Wang R, Brann D, Bieberich E and Wang G: Low dose Hsp90 inhibitor 17AAG protects neural progenitor cells from ischemia induced death. J Cell Commun Signal. 8:353–362. 2014. View Article : Google Scholar : PubMed/NCBI
17	Chinese Society of Neurology and Chinese Stroke Society, . Chinese guidelines for diagnosis and treatment of acute ischemic stroke 2018. Chin J Neurol. 9:666–682. 2018.
18	Adelson JD, Barreto GE, Xu L, Kim T, Brott BK, Ouyang YB, Naserke T, Djurisic M, Xiong X, Shatz CJ and Giffard RG: Neuroprotection from stroke in the absence of MHCI or PirB. Neuron. 73:1100–1107. 2012. View Article : Google Scholar : PubMed/NCBI
19	Pundik S, Xu K and Sundararajan S: Reperfusion brain injury: Focus on cellular bioenergetics. Neurology. 79 (13 Suppl 1):S44–S51. 2012. View Article : Google Scholar : PubMed/NCBI
20	Zhang Z, Yan J, Taheri S, Liu KJ and Shi H and Shi H: Hypoxia-inducible factor 1 contributes to N-acetylcysteine's protection in stroke. Free Radic Biol Med. 68:8–21. 2014. View Article : Google Scholar : PubMed/NCBI
21	Wen XR, Li C, Zong YY, Yu CZ, Xu J, Han D and Zhang GY: Dual inhibitory roles of geldanamycin on the c-Jun NH2-terminal kinase 3 signal pathway through suppressing the expression of mixed-lineage kinase 3 and attenuating the activation of apoptosis signal-regulating kinase 1 via facilitating the activation of Akt in ischemic brain injury. Neuroscience. 156:483–497. 2008. View Article : Google Scholar : PubMed/NCBI
22	Qi J, Liu Y, Yang P, Chen T, Liu XZ, Yin Y, Zhang J and Wang F: Heat shock protein 90 inhibition by 17-dimethylaminoethylamino-17-demethoxygeldanamycin protects blood-brain barrier integrity in cerebral ischemic stroke. Am J Transl Res. 7:1826–1837. 2015.PubMed/NCBI
23	Qi J, Han X, Liu HT, Chen T, Zhang JL, Yang P, Bo SH, Lu XT and Zhang J: 17-Dimethylaminoethylamino-17-demethoxygel-danamycin attenuates inflammatory responses in experimental stroke. Biol Pharm Bull. 37:1713–1718. 2014. View Article : Google Scholar : PubMed/NCBI
24	Voss AK, Thomas T and Gruss P: Mice lacking HSP90beta fail to develop a placental labyrinth. Development. 127:1–11. 2000. View Article : Google Scholar : PubMed/NCBI
25	Pajarillo E, Rizor A, Lee J, Aschner M and Lee E: The role of astrocytic glutamate transporters GLT-1 and GLAST in neurological disorders: Potential targets for neurotherapeutics. Neuropharmacology. 161:1075592019. View Article : Google Scholar : PubMed/NCBI
26	Mahmoud S, Gharagozloo M, Simard C and Gris D: Astrocytes maintain glutamate homeostasis in the CNS by controlling the balance between glutamate uptake and release. Cells. 8:1842019. View Article : Google Scholar : PubMed/NCBI
27	Wang H, Li J, Zhang H, Wang M, Xiao L, Wang Y and Cheng Q: Regulation of microglia polarization after cerebral ischemia. Front Cell Neurosci. 17:11826212023. View Article : Google Scholar : PubMed/NCBI
28	Leung SW, Lai JH, Wu JC, Tsai YR, Chen YH, Kang SJ, Chiang YH, Chang CF and Chen KY: Neuroprotective effects of emodin against ischemia/reperfusion injury through activating ERK-1/2 signaling pathway. Int J Mol Sci. 21:28992020. View Article : Google Scholar : PubMed/NCBI
29	Peng M, Ling X, Song R, Gao X, Liang Z, Fang F and Cang J: Upregulation of GLT-1 via PI3K/Akt pathway contributes to neuroprotection induced by dexmedetomidine. Front Neurol. 10:10412019. View Article : Google Scholar : PubMed/NCBI
30	Bacigaluppi M, Russo GL, Peruzzotti-Jametti L, Rossi S, Sandrone S, Butti E, De Ceglia R, Bergamaschi A, Motta C, Gallizioli M, et al: Neural stem cell transplantation induces stroke recovery by upregulating glutamate transporter GLT-1 in astrocytes. J Neurosci. 36:10529–10544. 2016. View Article : Google Scholar : PubMed/NCBI
31	Krzyżanowska W, Pomierny B, Bystrowska B, Pomierny-Chamioło L, Filip M, Budziszewska B and Pera J: Ceftriaxone- and N-acetylcysteine-induced brain tolerance to ischemia: Influence on glutamate levels in focal cerebral ischemia. PLoS One. 12:e01862432017. View Article : Google Scholar : PubMed/NCBI
32	Sha L, Wang X, Li J, Shi X, Wu L, Shen Y and Xu Q: Pharmacologic inhibition of Hsp90 to prevent GLT-1 degradation as an effective therapy for epilepsy. J Exp Med. 214:547–563. 2017. View Article : Google Scholar : PubMed/NCBI
33	Peng YC, Wang S, Zhang Y, Huang LJ, Wang XL and Peng Y: Hsp90β inhibitors prevent GLT-1 degradation but have no beneficial efficacy on absence epilepsy. J Asian Nat Prod Res. 21:905–915. 2019. View Article : Google Scholar : PubMed/NCBI