Effects of thymosin β4 on neuronal apoptosis in a rat model of cerebral ischemia‑reperfusion injury

Zhang,Zhongsheng; Liu,Shuangfeng; Huang,Sichun

doi:10.3892/mmr.2019.10683

Effects of thymosin β4 on neuronal apoptosis in a rat model of cerebral ischemia‑reperfusion injury

Authors:
- Zhongsheng Zhang
- Shuangfeng Liu
- Sichun Huang
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Affiliations: Department of Neurology, The Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital, Qingyuan, Guangdong 511518, P.R. China
Published online on: September 13, 2019 https://doi.org/10.3892/mmr.2019.10683
Pages: 4186-4192
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to investigate the protective effects of thymosin β4 (Tβ4) on neuronal apoptosis in rat middle cerebral artery occlusion ischemia/reperfusion (MCAO I/R) injury, and determine the mechanisms involved in this process. Forty‑eight adult male Sprague‑Dawley rats were randomly divided into three groups (n=16 per group): A sham control group, an ischemia/reperfusion group (I/R group), and a Tβ4 group. The focal cerebral I/R model was established by blocking the right MCA for 2 h, followed by reperfusion for 24 h. The Zea‑Longa method was used to assess neurological deficits. Cerebral infarct volume was assessed using 2,3,5‑triphenyltetrazolium chloride staining, and pathological changes were observed via hematoxylin and eosin staining. The terminal dexynucleotidyl transferase (TdT)‑mediated dUTP nick end labeling (TUNEL) assay was used to detect apoptosis. The expression of glucose‑regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), and caspase‑12 (CASP12) protein was assessed using immunohistochemistry and western blotting 24 h after reperfusion. Infarct volume and neuronal damage in the I/R and Tβ4 groups were significantly greater than those observed in the sham group. The Zea‑Longa score, neuronal apoptosis, and expression of GRP78, CHOP, and CASP12 in the I/R and Tβ4 groups were significantly higher than those reported in the sham group. However, the Longa score and neuronal apoptosis were lower in the Tβ4 group compared to the I/R group. The expression of GRP78 was significantly increased, whereas that of CHOP and CASP12 was significantly decreased in the Tβ4 group compared to the I/R group. The present data revealed that Tβ4 can inhibit neuronal apoptosis by upregulating GRP78 and downregulating CHOP and CASP12, thereby reducing cerebral I/R injury.

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1	GBD 2013 Mortailty and Cause of Death Collaborators, . Global, regional, and national age-sex specific all-cause and cause-specific mortality for 240 causes of death, 1990–2013: A systematic analysis for the global burden of disease study 2013. Lancet. 385:117–171. 2015. View Article : Google Scholar : PubMed/NCBI
2	Lai T, Li M, Zheng L, Song Y, Xu X, Guo Y, Zhang Y, Zhang Z and Mei Y: Over-expression of VEGF in marrow stromal cells promotes angiogenesis in rats with cerebral infarction via the synergistic effects of VEGF and Ang-2. J Huazhong Uni Sci Technolog Med Sci. 32:724–731. 2012. View Article : Google Scholar
3	Benjamin EJ, Muntner P, Alonso A, Bittencourt MS, Callaway CW, Carson AP, Chamberlain AM, Chang AR, Cheng S, Das SR, et al: Heart disease and stroke statistics-2019 update: A report from the american heart association. Circulation. 139:e56–e528. 2019. View Article : Google Scholar : PubMed/NCBI
4	Liberale L, Carbone F, Montecucco F, Gebhard C, Lüscher TF, Wegener S and Camici GG: Ischemic stroke across sexes: What is the status quo? Front Neuroendocrinol. 50:3–17. 2018. View Article : Google Scholar : PubMed/NCBI
5	Ma XH, Gao Q, Jia Z and Zhang ZW: Neuroprotective capabilities of TSA against cerebral ischemia/reperfusion injury via PI3K/Akt signaling pathway in rats. Int J Neurosci. 125:140–146. 2015. View Article : Google Scholar : PubMed/NCBI
6	Dong S, Tong X, Li J, Huang C, Hu C, Jiao H and Gu Y: Total flavonoid of Litsea coreana leve exerts anti-oxidative effects and alleviates focal cerebral ischemia/reperfusion injury. Neural Regen Res. 8:3193–3202. 2013.PubMed/NCBI
7	Thompson JW, Narayanan SV, Koronowski KB, Morris- Blanco K, Dave KR and Perez-Pinzon MA: Signaling pathways leading to ischemic mitochondrial neuroprotection. J Bioenerg Biomembr. 47:101–110. 2015. View Article : Google Scholar : PubMed/NCBI
8	Sadana P, Coughlin L, Burke J, Woods R and Mdzinarishvili A: Anti-edema action of thyroid hormone in MCAO model of ischemic brain stroke: Possible association with AQP4 modulation. J Neurol Sci. 354:37–45. 2015. View Article : Google Scholar : PubMed/NCBI
9	Zhang K: Integration of ER stress, oxidative stress and the inflammatory response in health and disease. Int J Clin Exp Med. 3:33–40. 2010.PubMed/NCBI
10	Shen YQ, Guerra-Librero A, Fernandez-Gil BI, Florido J, García-López S, Martinez-Ruiz L, Mendivil-Perez M, Soto-Mercado V, Acuña-Castroviejo D, Ortega-Arellano H, et al: Combination of melatonin and rapamycin for head and neck cancer therapy: Suppression of AKT/mTOR pathway activation and activation of mitophagy and apoptosis via mitochondrial function regulation. J Pineal Res. 64:2018. View Article : Google Scholar : PubMed/NCBI
11	Cybulsky AV: Endoplasmic reticulum stress, the unfolded protein response and autophagy in kidney diseases. Nat Rev Nephrol. 13:681–696. 2017. View Article : Google Scholar : PubMed/NCBI
12	Goldstein AL, Hannappel E and Kleinman HK: Thymosin beta4: Actin-sequestering protein moonlights to repair injured tissues. Trends Mol Med. 11:421–429. 2005. View Article : Google Scholar : PubMed/NCBI
13	Wong CG, Taban M, Osann K, Ross-Cisneros FN, Bruice TC, Zahn G and You T: Subchoroidal release of VEGF and bFGF produces choroidal neovascularization in rabbit. Curr Eye Res. 42:237–243. 2017. View Article : Google Scholar : PubMed/NCBI
14	Kim S and Kwon J: Thymosin beta 4 improves dermal burn wound healing via downregulation of receptor of advanced glycation end products in db/db mice. Biochim Biophys Acta. 1840:3452–3459. 2014. View Article : Google Scholar : PubMed/NCBI
15	Sosne G and Ousler GW: Thymosin beta 4 ophthalmic solution for dry eye: A randomized, placebo-controlled, phase II clinical trial conducted using the controlled adverse environment (CAE™) model. Clin Ophthalmol. 9:877–884. 2015.PubMed/NCBI
16	Choi SY, Noh MR, Kim DK, Sun W and Kim H: Neuroprotective function of thymosin-beta and its derivative peptides on the programmed cell death of chick and rat neurons. Biochem Biophys Res Commun. 362:587–593. 2007. View Article : Google Scholar : PubMed/NCBI
17	Morris DC, Cui Y, Cheung WL, Lu M, Zhang L, Zhang ZG and Chopp M: A dose-response study of thymosin β4 for the treatment of acute stroke. J Neurol Sci. 345:61–67. 2014. View Article : Google Scholar : PubMed/NCBI
18	Liu XH, Bi HY, Cao J, Ren S and Yue SW: Early constraint-induced movement therapy affects behavior and neuronal plasticity in ischemia-injured rat brains. Neural Regen Res. 14:775–782. 2019. View Article : Google Scholar : PubMed/NCBI
19	O'Donnell ME, Tran L, Lam TI, Liu XB and Anderson SE: Bumetanide inhibition of the blood-brain barrier Na-K-Cl cotransporter reduces edema formation in the rat middle cerebral artery occlusion model of stroke. J Cereb Blood Flow Metab. 24:1046–1056. 2004. View Article : Google Scholar : PubMed/NCBI
20	Li J, Yang S and Zhu G: Postnatal calpain inhibition elicits cerebellar cell death and motor dysfunction. Oncotarget. 8:87997–88007. 2017.PubMed/NCBI
21	Song ZJ, Yang SJ, Han L, Wang B and Zhu G: Postnatal calpeptin treatment causes hippocampal neurodevelopmental defects in neonatal rats. Neural Regen Res. 14:834–840. 2019. View Article : Google Scholar : PubMed/NCBI
22	Jiang S, Li T, Ji T, Yi W, Yang Z, Wang S, Yang Y and Gu C: AMPK: Potential therapeutic target for ischemic stroke. Theranostics. 8:4535–4551. 2018. View Article : Google Scholar : PubMed/NCBI
23	Lv J, Jiang S, Yang Z, Hu W, Wang Z, Li T and Yang Y: PGC-1α sparks the fire of neuroprotection against neurodegenerative disorders. Ageing Res Rev. 44:8–21. 2018. View Article : Google Scholar : PubMed/NCBI
24	Kuzan A: Thymosin β as an actin-binding protein with a variety of functions. Adv Clin Exp Med. 25:1331–1336. 2016. View Article : Google Scholar : PubMed/NCBI
25	Wang YY, Zhu QS, Wang YW and Yin RF: Thymosin beta-4 recombinant adeno-associated virus enhances human nucleus pulposus cell proliferation and reduces cell apoptosis and senescence. Chin Med J (Engl). 128:1529–1535. 2015. View Article : Google Scholar : PubMed/NCBI
26	Sosne G, Rimmer D, Kleinman HK and Ousler G: Thymosin beta 4: A potential novel therapy for neurotrophic keratopathy, dry eye, and ocular surface diseases. Vitam Horm. 102:277–306. 2016. View Article : Google Scholar : PubMed/NCBI
27	Stark CK, Tarkia M, Kentala R, Malmberg M, Vähäsilta T, Savo M, Hynninen VV, Helenius M, Ruohonen S, Jalkanen J, et al: Systemic dosing of thymosin beta 4 before and after ischemia does not attenuate global myocardial ischemia-reperfusion injury in pigs. Front Pharmacol. 7:1152016. View Article : Google Scholar : PubMed/NCBI
28	Deville C, Girard-Blanc C, Assrir N, Nhiri N, Jacquet E, Bontems F, Renault L, Petres S and van Heijenoort C: Mutations in actin used for structural studies partially disrupt β-thymosin/WH2 domains interaction. FEBS Lett. 590:3690–3699. 2016. View Article : Google Scholar : PubMed/NCBI
29	Jiang W, Liang G, Li X, Li Z, Gao X, Feng S, Wang X, Liu M and Liu Y: Intracarotid transplantation of autologous adipose-derived mesenchymal stem cells significantly improves neurological deficits in rats after MCAo. J Mater Sci Mater Med. 25:1357–1366. 2014. View Article : Google Scholar : PubMed/NCBI
30	Yu ZH, Cai M, Xiang J, Zhang ZN, Zhang JS, Song XL, Zhang W, Bao J, Li WW and Cai DF: PI3K/Akt pathway contributes to neuroprotective effect of Tongxinluo against focal cerebral ischemia and reperfusion injury in rats. J Ethnopharmacol. 181:8–19. 2016. View Article : Google Scholar : PubMed/NCBI
31	Brown MK and Naidoo N: The endoplasmic reticulum stress response in aging and age-related diseases. Front Physiol. 3:2632012. View Article : Google Scholar : PubMed/NCBI
32	Hetz C: The unfolded protein response: Controlling cell fate decisions under ER stress and beyond. Nat Rev Mol Cell Biol. 13:89–102. 2012. View Article : Google Scholar : PubMed/NCBI
33	Li J, Ni M, Lee B, Barron E, Hinton DR and Lee AS: The unfolded protein response regulator GRP78/BiP is required for endoplasmic reticulum integrity and stress-induced autophagy in mammalian cells. Cell Death Differ. 15:1460–1471. 2008. View Article : Google Scholar : PubMed/NCBI
34	Ye Z, Wang N, Xia P, Wang E, Liao J and Guo Q: Parecoxib suppresses CHOP and Foxo1 nuclear translocation, but increases GRP78 levels in a rat model of focal ischemia. Neurochemical Res. 38:686–693. 2013. View Article : Google Scholar
35	Chen HL, Qi H, Liu XJ and Wang MS: Effect of electroacupuncture pretreatment on apoptotic neurons and expression of GRP 78 and GADD 153 in the hippocampus in rats with global cerebral ischemia/reperfusion injury. Zhen Ci Yan Jiu. 39:431–436. 2014.(In Chinese). PubMed/NCBI
36	Dong YF, Chen ZZ, Zhao Z, Yang DD, Yan H, Ji J and Sun XL: Potential role of microRNA-7 in the anti-neuroinflammation effects of nicorandil in astrocytes induced by oxygen-glucose deprivation. J Neuroinflammation. 13:602016. View Article : Google Scholar : PubMed/NCBI
37	Tong Q, Wu L, Jiang T, Ou Z, Zhang Y and Zhu D: Inhibition of endoplasmic reticulum stress-activated IRE1α-TRAF2-caspase-12 apoptotic pathway is involved in the neuroprotective effects of telmisartan in the rotenone rat model of Parkinson's disease. Eur J Pharmacol. 776:106–115. 2016. View Article : Google Scholar : PubMed/NCBI
38	Zhang Q, Liu J, Chen S, Liu J, Liu L, Liu G, Wang F, Jiang W, Zhang C, Wang S and Yuan X: Caspase-12 is involved in stretch-induced apoptosis mediated endoplasmic reticulum stress. Apoptosis. 21:432–442. 2016. View Article : Google Scholar : PubMed/NCBI