TIMP3 attenuates cerebral ischemia/reperfusion‑induced apoptosis and oxidative stress in neurocytes by regulating the AKT pathway

Meng,Linglei; Zhang,Yongting; Li,Demao; Shang,Xinfang; Hao,Xuejia; Chen,Xin; Gao,Fengxiao

doi:10.3892/etm.2021.10405

TIMP3 attenuates cerebral ischemia/reperfusion‑induced apoptosis and oxidative stress in neurocytes by regulating the AKT pathway

Authors:
- Linglei Meng
- Yongting Zhang
- Demao Li
- Xinfang Shang
- Xuejia Hao
- Xin Chen
- Fengxiao Gao
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Affiliations: Department of Imaging, Xingtai People's Hospital, Xingtai, Hebei 054001, P.R. China, Department of Imaging, Xingtai Orthopedic Hospital, Xingtai, Hebei 054001, P.R. China, Department of Cardiothoracic Surgery, Xingtai People's Hospital, Xingtai, Hebei 054001, P.R. China, Department of Neurology, Xingtai People's Hospital, Xingtai, Hebei 054001, P.R. China
Published online on: July 8, 2021 https://doi.org/10.3892/etm.2021.10405
Article Number: 973
Copyright: © Meng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Ischemic stroke seriously threatens human health and creates a large social burden. The present study investigated whether tissue inhibitor of metalloproteinases‑3 (TIMP3) prevented cerebral ischemia/reperfusion (I/R), with the aim to explore the underlying mechanism. A transient middle cerebral artery occlusion model was conducted in mice, and oxygen glucose deprivation and reoxygenation (OGD/R) was investigated in PC12 cells to mimic cerebral ischemia‑reperfusion injury (CIRI). Western blotting was used to determine the expression of TIMP3, Bax, Bcl‑2 and AKT. TUNEL was used to detect apoptosis in cerebral tissues or cultured PC12 cells. Expression levels of reactive oxygen species (ROS), superoxide dismutase (SOD) and malondialdehyde (MDA) were detected to reveal oxidative stress. The results demonstrated that TIMP3 expression was significantly decreased after I/R in vivo or OGD/R in vitro, and the number of TUNEL‑positive cells was reduced by the overexpression of TIMP3. The attenuation of Bax/Bcl‑2 ratio in OGD/R‑induced PC12 cells suppressed the expression levels of ROS and MDA; while also elevating SOD activity in the OGD/R‑induced neurocytes in vitro. In addition, TIMP3‑overexpression reversed the downregulation of phosphorylated‑AKT (Thr308 and Ser473) in OGD/R‑treated PC12 cells. However, the anti‑apoptotic and anti‑oxidative stress roles of TIMP3 in OGD/R‑induced PC12 cells were partially abolished after treatment with the AKT inhibitor, AZD5363. Overall, TIMP3 exerted an anti‑apoptotic and anti‑oxidative stress role in CIRI through the AKT pathway, which may be a potential therapeutic target for the treatment of CIRI.

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1	Bosetti F, Koenig JI, Ayata C, Back SA, Becker K, Broderick JP, Carmichael ST, Cho S, Cipolla MJ, Corbett D, et al: Translational stroke research: Vision and opportunities. Stroke. 48:2632–2637. 2017.PubMed/NCBI View Article : Google Scholar
2	Dorado L, Millan M and Davalos A: Reperfusion therapies for acute ischemic stroke: An update. Curr Cardiol Rev. 10:327–335. 2014.PubMed/NCBI View Article : Google Scholar
3	Nishijima Y, Akamatsu Y, Weinstein PR and Liu J: Collaterals: Implications in cerebral ischemic diseases and therapeutic interventions. Brain Res. 1623:18–29. 2015.PubMed/NCBI View Article : Google Scholar
4	Zhang J, Fang X, Zhou Y, Deng X, Lu Y, Li J, Li S, Wang B and Xu R: The possible damaged mechanism and the preventive effect of monosialotetrahexosylganglioside in a rat model of cerebral ischemia-reperfusion injury. J Stroke Cerebrovasc Dis. 24:1471–1478. 2015.PubMed/NCBI View Article : Google Scholar
5	Liang G, Shi B, Luo W and Yang J: The protective effect of caffeic acid on global cerebral ischemia-reperfusion injury in rats. Behav Brain Funct. 11(18)2015.PubMed/NCBI View Article : Google Scholar
6	Siroli L, Braschi G, de Jong A, Kok J, Patrignani F and Lanciotti R: Transcriptomic approach and membrane fatty acid analysis to study the response mechanisms of Escherichia coli to thyme essential oil, carvacrol, 2-(E)-hexanal and citral exposure. J Appl Microbiol. 125:1308–1320. 2018.PubMed/NCBI View Article : Google Scholar
7	Di Gregoli K, Mohamad Anuar NN, Bianco R, White SJ, Newby AC, George SJ and Johnson JL: MicroRNA-181b controls atherosclerosis and aneurysms through regulation of TIMP-3 and elastin. Circ Res. 120:49–65. 2017.PubMed/NCBI View Article : Google Scholar
8	Brew K and Nagase H: The tissue inhibitors of metalloproteinases (TIMPs): An ancient family with structural and functional diversity. Biochim Biophys Acta. 1803:55–71. 2010.PubMed/NCBI View Article : Google Scholar
9	Shen B, Jiang Y, Chen YR, Zheng HC, Zeng W, Li YY, Yin A and Nie Y: Expression and inhibitory role of TIMP-3 in hepatocellular carcinoma. Oncol Rep. 36:494–502. 2016.PubMed/NCBI View Article : Google Scholar
10	Kubatka P, Uramova S, Kello M, Kajo K, Kruzliak P, Mojzis J, Vybohova D, Adamkov M, Jasek K, Lasabova Z, et al: Antineoplastic effects of clove buds (Syzygium aromaticum L.) in the model of breast carcinoma. J Cell Mol Med. 21:2837–2851. 2017.PubMed/NCBI View Article : Google Scholar
11	Liu H, Jing X, Dong A, Bai B and Wang H: Overexpression of TIMP3 protects against cardiac ischemia/reperfusion injury by inhibiting myocardial apoptosis through ROS/Mapks pathway. Cell Physiol Biochem. 44:1011–1023. 2017.PubMed/NCBI View Article : Google Scholar
12	Menghini R, Casagrande V, Menini S, Marino A, Marzano V, Hribal ML, Gentileschi P, Lauro D, Schillaci O, Pugliese G, et al: TIMP3 overexpression in macrophages protects from insulin resistance, adipose inflammation, and nonalcoholic fatty liver disease in mice. Diabetes. 61:454–462. 2012.PubMed/NCBI View Article : Google Scholar
13	Fujii T, Duarte S, Lee E, Ke B, Busuttil RW and Coito AJ: Tissue inhibitor of metalloproteinase 3 deficiency disrupts the hepatocyte E-cadherin/β-catenin complex and induces cell death in liver ischemia/reperfusion injury. Liver Transpl. 26:113–126. 2020.PubMed/NCBI View Article : Google Scholar
14	Wang G, Wang T, Hu Y, Wang J, Wang Y, Zhang Y, Li F, Liu W, Sun Y, Yu B and Kou J: NMMHC IIA triggers neuronal autophagic cell death by promoting F-actin-dependent ATG9A trafficking in cerebral ischemia/reperfusion. Cell Death Dis. 11(428)2020.PubMed/NCBI View Article : Google Scholar
15	Zhou Z, Xu N, Matei N, McBride DW, Ding Y, Liang H, Tang J and Zhang JH: Sodium butyrate attenuated neuronal apoptosis via GPR41/Gβγ/PI3K/Akt pathway after MCAO in rats. J Cereb Blood Flow Metab. 41:267–281. 2021.PubMed/NCBI View Article : Google Scholar
16	Li Y, Guo S, Liu W, Jin T, Li X, He X, Zhang X, Su H, Zhang N and Duan C: Silencing of SNHG12 enhanced the effectiveness of MSCs in alleviating ischemia/reperfusion injuries via the PI3K/AKT/mTOR signaling pathway. Front Neurosci. 13(645)2019.PubMed/NCBI View Article : Google Scholar
17	Feng H, Hu L, Zhu H, Tao L, Wu L, Zhao Q, Gao Y, Gong Q, Mao F, Li X, et al: Repurposing antimycotic ciclopirox olamine as a promising anti-ischemic stroke agent. Acta Pharm Sin B. 10:434–446. 2020.PubMed/NCBI View Article : Google Scholar
18	Gibb SL, Zhao Y, Potter D, Hylin MJ, Bruhn R, Baimukanova G, Zhao J, Xue H, Abdel-Mohsen M, Pillai SK, et al: TIMP3 attenuates the loss of neural stem cells, mature neurons and neurocognitive dysfunction in traumatic brain injury. Stem Cells. 33:3530–3544. 2015.PubMed/NCBI View Article : Google Scholar
19	Bu X, Zhang N, Yang X, Liu Y, Du J, Liang J, Xu Q and Li J: Proteomic analysis of cPKCβII-interacting proteins involved in HPC-induced neuroprotection against cerebral ischemia of mice. J Neurochem. 117:346–356. 2011.PubMed/NCBI View Article : Google Scholar
20	Leary S, Anthony R, Grandin T, Greenacre C, Gwaltney-Brant S, Ann McCrackin M, Meyer R, Miller D, Shearer J, Turner T and Yanong R: AVMA guidelines for the euthanasia of animals: 2020 Edition*. AVMA, 2020.
21	Liu X, Li M, Hou M, Huang W and Song J: MicroRNA-135a alleviates oxygen-glucose deprivation and reoxygenation-induced injury in neurons through regulation of GSK-3β/Nrf2 signaling. J Biochem Mol Toxicol: e22159, 2018. doi: 10.1002/jbt.22159.
22	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.PubMed/NCBI View Article : Google Scholar
23	Broughton BR, Reutens DC and Sobey CG: Apoptotic mechanisms after cerebral ischemia. Stroke. 40:e331–e339. 2009.PubMed/NCBI View Article : Google Scholar
24	Li P, Shen M, Gao F, Wu J, Zhang J, Teng F and Zhang C: An antagomir to microRNA-106b-5p ameliorates cerebral ischemia and reperfusion injury in rats via inhibiting apoptosis and oxidative stress. Mol Neurobiol. 54:2901–2921. 2017.PubMed/NCBI View Article : Google Scholar
25	Woodruff TM, Thundyil J, Tang SC, Sobey CG, Taylor SM and Arumugam TV: Pathophysiology, treatment, and animal and cellular models of human ischemic stroke. Mol Neurodegener. 6(11)2011.PubMed/NCBI View Article : Google Scholar
26	Lewen A, Matz P and Chan PH: Free radical pathways in CNS injury. J Neurotrauma. 17:871–890. 2000.PubMed/NCBI View Article : Google Scholar
27	Liu X, Qing Wang, Cui Y, Li X and Yang H: In-depth transcriptomic and proteomic analyses of the hippocampus and cortex in a rat model after cerebral ischemic injury and repair by Shuxuetong (SXT) injection. J Ethnopharmacol. 249(112362)2019.PubMed/NCBI View Article : Google Scholar
28	Barialai L, Strecker MI, Luger AL, Jäger M, Bruns I, Sittig ACM, Mildenberger IC, Heller SM, Delaidelli A, Lorenz NI, et al: AMPK activation protects astrocytes from hypoxia-induced cell death. Int J Mol Med. 45:1385–1396. 2020.PubMed/NCBI View Article : Google Scholar