Activation of autophagy in rat brain cells following focal cerebral ischemia reperfusion through enhanced expression of Atg1/pULK and LC3

Yu,Jingwei; Bao,Cuifen; Dong,Yanru; Liu,Xia

doi:10.3892/mmr.2015.3850

Activation of autophagy in rat brain cells following focal cerebral ischemia reperfusion through enhanced expression of Atg1/pULK and LC3

Authors:
- Jingwei Yu
- Cuifen Bao
- Yanru Dong
- Xia Liu
View Affiliations

Affiliations: Department of Histology and Embryology, Liaoning Medical University, Jinzhou, Liaoning 121001, P.R. China, Key Laboratory of Molecular Cell Biology and New Drug Development, Liaoning Medical University, Jinzhou, Liaoning 121001, P.R. China
Published online on: May 26, 2015 https://doi.org/10.3892/mmr.2015.3850
Pages: 3339-3344
Copyright: © Yu et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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

The present study aimed to investigate the activation of Atg1/pULK, and LC3 in the cerebral cortex following focal cerebral ischemia reperfusion (CIR) injury, thereby examining its effect on autophagy in brain cells. Rat CIR models were established using the technique of middle cerebral artery occlusion. The neurological function score, TTC staining and the water content of brain tissue were used to evaluate the CIR model. Levels of autophagy in the brain cells were examined at different time‑points following CIR damage using electron microscopy. Immunohistochemistry and western blot analysis were also used for the qualitative and quantitative detection of levels of Atg1/pULK and LC3 in the cerebral cortex. Autophagy was observed in the early stage of CIR, and the expression of Atg1/pULK and LC3 were observed 1 h following CIR in the rats and reached peak expression levels after12 h, which following which the they gradually decreased. These results suggested Atg1/pULK and LC3 are key in the regulation of autophagy following CIR in the rat brain.

View Figures

View References

1	Hankey GJ, Jamrozik K, Broadhurst RJ, Forbes S, Burvill PW, Anderson CS, et al: Five-year survival after first-ever stroke and related prognostic factors in the Perth community stroke study. Stroke. 31:2080–2086. 2000. View Article : Google Scholar : PubMed/NCBI
2	Gosman-Hedstrom G, Claesson L, Klingenstierna U, Carlsson J, Olausson B, Frizell M, et al: Effects of acupuncture treatment on daily life activities and quality of life: A controlled, prospective and randomized study of acute stroke patients. Stroke. 29:2100–2108. 1998. View Article : Google Scholar
3	Naeser MA and Hamblin MR: Potential for transcranial laser or LED therapy to treat stroke, traumatic brain injury and neurodegenerative disease. Photomed Laser Surg. 29:443–446. 2011. View Article : Google Scholar : PubMed/NCBI
4	Minnerup J and Schabitz WR: Improving outcome after stroke: time to treat new targets. Stroke. 43:295–296. 2012. View Article : Google Scholar
5	Koike M, Shibata M, Tadakoshi M, Gotoh K, Komatsu M, Waguri S, et al: Inhibition of autophagy prevents hippocampal pyramidal neuron death after hypoxic-ischemic injury. Am J Pathol. 172:454–469. 2008. View Article : Google Scholar : PubMed/NCBI
6	Fan J, Zhang Z, Chao X, Gu J, Cai W, Zhou W, et al: Ischemic preconditioning enhances autophagy but suppresses autophagic cell death in rat spinal neurons following ischemia-reperfusion. Brain Res. 1562:76–86. 2014. View Article : Google Scholar : PubMed/NCBI
7	Puyal J, Vaslin A, Mottier V and Clarke PG: Postischemic treatment of neonatal cerebral ischemia should target autophagy. Ann Neurol. 66:378–389. 2009. View Article : Google Scholar : PubMed/NCBI
8	Smith CM, Chen Y, Sullivan ML, Kochanek PM and Clark RS: Autophagy in acute brain injury: Feast, famine, or folly? Neurobiol Dis. 43:52–59. 2011. View Article : Google Scholar :
9	Puyal J, Ginet V, Grishchuk Y, Truttmann AC and Clarke PG: Neuronal autophagy as a mediator of life and death: Contrasting roles in chronic neurodegenerative and acute neural disorders. Neuroscientist. 18:224–236. 2012. View Article : Google Scholar
10	Ryter SW, Lee SJ, Smith A and Choi AM: Autophagy in vascular disease. Proc Am Thorac Soc. 7:40–47. 2010. View Article : Google Scholar : PubMed/NCBI
11	Jung G, Roh J, Lee H, Gil M, Yoon DH, Suh C, et al: Autophagic markers BECLIN 1 and LC3 are associated with prognosis of multiple myeloma. Acta Haematol. 134:17–24. 2015. View Article : Google Scholar : PubMed/NCBI
12	Wen YD, Sheng R, Zhang LS, Han R, Zhang X, et al: Neuronal injury in rat model of permanent focal cerebral ischemia is associated with activation of autophagic and lysosomal pathways. Autophagy. 4:762–769. 2008. View Article : Google Scholar : PubMed/NCBI
13	Rami A, Langhagen A and Steiger S: Focal cerebral ischemia induces upregulation of Beclin1 and autophagy-like cell death. Neurobiol Dis. 29:132–141. 2008. View Article : Google Scholar
14	Puyal J, Vaslin A, Mottier V and Clarke PG: Postischemic treatment of neonatal cerebral ischemia should target autophagy. Ann Neurol. 66:378–389. 2009. View Article : Google Scholar : PubMed/NCBI
15	Zheng YQ, Liu JX, Li XZ, Xu L and Xu YG: RNA interference-mediated downregulation of Beclin1 attenuates cerebral ischemic injury in rats. Acta Pharmocol Sin. 30:919–927. 2009. View Article : Google Scholar
16	Zhang X, Yan H, Yuan Y, Gao J, Shen Z, Cheng Y, et al: Cerebral ischemia-reperfusion-induced autophagy protects against neuronal injury by mitochondrial clearance. Autophagy. 9:1321–1333. 2013. View Article : Google Scholar : PubMed/NCBI
17	Hoehn BD, Palmer TD and Steinberg GK: Neurogenesis in rats after focal cerebral ischemia is enhanced by indomethacin. Stroke. 36:2718–2724. 2005. View Article : Google Scholar : PubMed/NCBI
18	Kofler J, Otsuka T, Zhang Z, Noppens R, Grafe MR, Koh DW, et al: Differential effect of PARP-2 deletion on brain injury after focal and global cerebral ischemia. J Cereb Blood Flow Metab. 26:135–141. 2006. View Article : Google Scholar
19	Kanno H, Ozawa H, Sekiguchi A and Itoi E: Spinal cord injury induces upregulation of Beclin 1 and promotes autophagic cell death. Neurobiol Dis. 33:143–148. 2009. View Article : Google Scholar
20	Grishchuk Y, Ginet V, Truttmann AC, Clarke PG and Puyal J: Beclin 1-independent autophagy contributes to apoptosis in cortical neurons. Autophagy. 7:1115–1131. 2011. View Article : Google Scholar : PubMed/NCBI
21	Rami A, Langhagen A and Steiger S: Focal cerebral ischemia induces upregulation of Beclin 1 and autophagy-like cell death. Neurobiol Dis. 29:132–141. 2008. View Article : Google Scholar
22	Chen X, Li M, Chen D, Gao W, Guan JL, Komatsu M, et al: Autophagy induced by calcium phosphate precipitates involves endoplasmic reticulum membranes in autophagosome biogenesis. PLoS One. 7:e523472012. View Article : Google Scholar
23	Koh PO: Melatonin attenuates the focal cerebral ischemic injury by inhibiting the dissociation of pBad from 14-3-3. J Pineal Res. 44:101–106. 2008.
24	Lafay-Chebassier C, Paccalin M, Page G, Barc-Pain S, Perault-Pochat MC, Gil R, et al: mTOR/p70S6k signalling alteration by Abeta exposure as well as in APP-PS1 transgenic models and in patients with Alzheimer's disease. J Neurochem. 94:215–225. 2005. View Article : Google Scholar : PubMed/NCBI
25	Meiling-Wesse K, Barth H, Voss C, Eskelinen EL, Epple UD and Thumm M: Atg21 is required for effective recruitment of Atg8 to the preautophagosomal structure during the Cvt pathway. J Biol Chem. 279:37741–37750. 2004. View Article : Google Scholar : PubMed/NCBI
26	Kofler J, Otsuka T, Zhang Z, Noppens R, Grafe MR, Koh DW, et al: Differential effect of PARP-2 deletion on brain injury after focal and global cerebral ischemia. J Cereb Blood Flow Metab. 26:135–141. 2006. View Article : Google Scholar
27	Tsubokawa T, Jadhav V, Solaroglu I, Shiokawa Y, Konishi Y and Zhang JH: Lecithinized superoxide dismutase improves outcomes and attenuates focal cerebral ischemic injury via anti-apoptotic mechanisms in rats. Stroke. 38:1057–1062. 2007. View Article : Google Scholar : PubMed/NCBI