Grape Seed Proanthocyanidins Protect N2a Cells against Ischemic Injury via Endoplasmic Reticulum Stress and Mitochondrial-associated Pathways

Author(s): Kun Fu, Liqiang Chen, Lifeng Miao*, Yan Guo, Wei Zhang, Yunan Bai*.

Journal Name: CNS & Neurological Disorders - Drug Targets
(Formerly Current Drug Targets - CNS & Neurological Disorders)

Volume 18 , Issue 4 , 2019

Become EABM
Become Reviewer

Graphical Abstract:


Abstract:

Background/Objective: Grape seed proanthocyanidins (GSPs) are a group of polyphenolic bioflavonoids, which possess a variety of biological functions and pharmacological properties. We studied the neuroprotective effects of GSP against oxygen-glucose deprivation/reoxygenation (OGD/R) injury and the potential mechanisms in mouse neuroblastoma N2a cells.

Methods: OGD/R was conducted in N2a cells. Cell viability was evaluated by CCK-8 and LDH release assay. Apoptosis was assessed by TUNEL staining and flow cytometry. Protein levels of cleaved caspase-3, Bax and Bcl-2 were detected by Western blotting. CHOP, GRP78 and caspase-12 mRNA levels were assessed by real-time PCR. JC-1 dying was used to detect mitochondrial membrane potential. ROS levels, activities of endogenous antioxidant enzymes and ATP production were examined to evaluate mitochondrial function.

Results: GSP increased cell viability after OGD/R injury in a dose-dependent manner. Furthermore, GSP inhibited cell apoptosis, reduced the mRNA levels of CHOP, GRP78 and caspase-12 (ER stressassociated genes), restored mitochondrial membrane potential and ATP generation, improved activities of endogenous anti-oxidant ability (T-AOC, GXH-Px, and SOD), and decreased ROS level.

Conclusion: Our findings suggest that GSP can protect N2a cells from OGD/R insult. The mechanism of anti-apoptotic effects of GSP may involve attenuating ER stress and mitochondrial dysfunction.

Keywords: Grape seed proanthocyanidins, oxygen-glucose deprivation/reoxygenation, apoptosis, endoplasmic reticulum stress, mitochondrial dysfunction, brain ischemia.

[1]
Su Y, Li F. Endoplasmic reticulum stress in brain ischemia. Int J Neurosci 2016; 126: 681-91.
[2]
Nguyen H, Zarriello S, Rajani M, Tuazon J, Napoli E, Borlongan CV. Understanding the role of dysfunctional and healthy mitochondria in stroke pathology and its treatment. Int J Mol Sci 2018; 19.
[3]
Bakthavachalam P, Shanmugam PST. Mitochondrial dysfunction - Silent killer in cerebral ischemia. J Neurol Sci 2017; 375: 417-23.
[4]
Mei Y, Thompson MD, Cohen RA, Tong X. Endoplasmic reticulum stress and related pathological processes. J Pharmacol Biomed Anal 2013; 11000107
[5]
Deniaud A. Sharaf el dein O, Maillier E, et al Endoplasmic reticulum stress induces calcium-dependent permeability transition, mitochondrial outer membrane permeabilization and apoptosis. Oncogene 2008; 27: 285-99.
[6]
Xu W, Liu L, Charles IG, Moncada S. Nitric oxide induces coupling of mitochondrial signalling with the endoplasmic reticulum stress response. Nat Cell Biol 2004; 6: 1129-34.
[7]
Wang S, Ma F, Huang L, et al. Dl-3-n-Butylphthalide (NBP): A promising therapeutic agent for ischemic stroke. CNS Neurol Disord Drug Targets 2018; 17: 338-47.
[8]
Borlongan CV, Nguyen H, Lippert T, et al. May the force be with you: Transfer of healthy mitochondria from stem cells to stroke cells. J Cereb Blood Flow Metab 2018: 271678X18811277
[9]
Tajiri N, Borlongan CV, Kaneko Y. Cyclosporine a treatment abrogates ischemia-induced neuronal cell death by preserving mitochondrial integrity through upregulation of the parkinson’s disease-associated protein DJ-1. CNS Neurosci Ther 2016; 22: 602-10.
[10]
Chu H, Tang Q, Huang H, Hao W, Wei X. Grape-seed proanthocyanidins inhibit the lipopolysaccharide-induced inflammatory mediator expression in RAW264.7 macrophages by suppressing MAPK and NF-kappab signal pathways. Environ Toxicol Pharmacol 2016; 41: 159-66.
[11]
Katiyar SK. Grape seed proanthocyanidines and skin cancer prevention: Inhibition of oxidative stress and protection of immune system. Mol Nutr Food Res 2008; 52(Suppl. 1): S71-6.
[12]
Yang X, Liu T, Chen B, Wang F, Yang Q, Chen X. Grape seed proanthocyanidins prevent irradiation-induced differentiation of human lung fibroblasts by ameliorating mitochondrial dysfunction. Sci Rep 2017; 7: 62.
[13]
Bashir N, Manoharan V, Miltonprabu S. Grape seed proanthocyanidins protects against cadmium induced oxidative pancreatitis in rats by attenuating oxidative stress, inflammation and apoptosis via Nrf-2/HO-1 signaling. J Nutr Biochem 2016; 32: 128-41.
[14]
Lu Z, Lu F, Zheng Y, Zeng Y, Zou C, Liu X. Grape seed proanthocyanidin extract protects human umbilical vein endothelial cells from indoxyl sulfate-induced injury via ameliorating mitochondrial dysfunction. Ren Fail 2016; 38: 100-8.
[15]
Zhang Z, Zheng L, Zhao Z, Shi J, Wang X, Huang J. Grape seed proanthocyanidins inhibit H2O2-induced osteoblastic MC3T3-E1 cell apoptosis via ameliorating H2O2-induced mitochondrial dysfunction. J Toxicol Sci 2014; 39: 803-13.
[16]
Wang X, Jia D, Zhang J, Wang W. Grape seed proanthocyanidins protect cardiomyocytes against hypoxia/reoxygenation injury by attenuating endoplasmic reticulum stress through PERK/eIF2alpha pathway. Mol Med Rep 2017; 16: 9189-96.
[17]
Ding Y, Dai X, Zhang Z, et al. Proanthocyanidins protect against early diabetic peripheral neuropathy by modulating endoplasmic reticulum stress. J Nutr Biochem 2014; 25: 765-72.
[18]
Ding Y, Zhang Z, Dai X, et al. Grape seed proanthocyanidins ameliorate pancreatic beta-cell dysfunction and death in low-dose streptozotocin- and high-carbohydrate/high-fat diet-induced diabetic rats partially by regulating endoplasmic reticulum stress. Nutr Metab 2013; 10: 51.
[19]
Ding Y, Dai X, Jiang Y, et al. Grape seed proanthocyanidin extracts alleviate oxidative stress and ER stress in skeletal muscle of low-dose streptozotocin- and high-carbohydrate/high-fat diet-induced diabetic rats. Mol Nutr Food Res 2013; 57: 365-9.
[20]
Kong X, Guan J, Gong S, Wang R. Neuroprotective effects of grape seed procyanidin extract on ischemia-reperfusion brain injury. Chin Med Sci J 2017; 32: 92-9.
[21]
Cao WL, Huang HB, Fang L, Hu JN, Jin ZM, Wang RW. Protective effect of ginkgo proanthocyanidins against cerebral ischemia/reperfusion injury associated with its antioxidant effects. Neural Regen Res 2016; 11: 1779-83.
[22]
Zhao XL, Yu CZ. Vosaroxin induces mitochondrial dysfunction and apoptosis in cervical cancer HeLa cells: Involvement of AMPK/Sirt3/HIF-1 pathway. Chem Biol Interact 2018; 290: 57-63.
[23]
Lu Y, Huang Z, Hua Y, Xiao G. Minocycline Promotes BDNF Expression of N2a Cells via inhibition of miR-155-mediated repression after oxygen-glucose deprivation and reoxygenation. Cell Mol Neurobiol 2018; 38: 1305-13.
[24]
Huang Y, Hu Z. UBIAD1 protects against oxygen-glucose deprivation/reperfusion-induced multiple subcellular organelles injury through PI3K/AKT pathway in N2A cells. J Cell Physiol 2018; 233: 7480-96.
[25]
Prentice H, Modi JP, Wu JY. Mechanisms of neuronal protection against excitotoxicity, endoplasmic reticulum stress, and mitochondrial dysfunction in stroke and neurodegenerative diseases. Oxid Med Cell Longev 2015; 2015964518
[26]
Schonthal AH. Endoplasmic reticulum stress: Its role in disease and novel prospects for therapy. Scientifica (Cairo) 2012; 2012857516
[27]
Ron D, Walter P. Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol 2007; 8: 519-29.
[28]
Lee AS. The ER chaperone and signaling regulator GRP78/BiP as a monitor of endoplasmic reticulum stress. Methods 2005; 35: 373-81.
[29]
Oyadomari S, Mori M. Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ 2004; 11: 381-9.
[30]
Oyadomari S, Koizumi A, Takeda K, et al. Targeted disruption of the Chop gene delays endoplasmic reticulum stress-mediated diabetes. J Clin Invest 2002; 109: 525-32.
[31]
McCullough KD, Martindale JL, Klotz LO, Aw TY, Holbrook NJ. Gadd153 sensitizes cells to endoplasmic reticulum stress by down-regulating Bcl2 and perturbing the cellular redox state. Mol Cell Biol 2001; 21: 1249-59.
[32]
Garcia de la Cadena S, Massieu L. Caspases and their role in inflammation and ischemic neuronal death. Focus on caspase-12. Apoptosis 2016; 21: 763-77.
[33]
Nakagawa T, Zhu H, Morishima N, et al. Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-beta. Nature 2000; 403: 98-103.
[34]
Kumar A, Dhawan A, Kadam A, Shinde A. Autophagy and mitochondria: Targets in neurodegenerative disorders. CNS Neurol Disord Drug Targets 2018; 17: 696-705.
[35]
Arola-Arnal A, Oms-Oliu G, Crescenti A, et al. Distribution of grape seed flavanols and their metabolites in pregnant rats and their fetuses. Mol Nutr Food Res 2013; 57: 1741-52.
[36]
Margalef M, Pons Z, Bravo FI, Muguerza B, Arola-Arnal A. Tissue distribution of rat flavanol metabolites at different doses. J Nutr Biochem 2015; 26: 987-95.


Rights & PermissionsPrintExport Cite as

Article Details

VOLUME: 18
ISSUE: 4
Year: 2019
Page: [334 - 341]
Pages: 8
DOI: 10.2174/1871527318666190212111650
Price: $58

Article Metrics

PDF: 46
HTML: 4

Special-new-year-discount