Enhancement of O-GlcNAcylation on Mitochondrial Proteins with 2-(4-Methoxyphenyl)Ethyl-2-Acetamido-2-Deoxy-β-d-Pyranoside, Contributes to the Mitochondrial Network, Cellular Bioenergetics and Stress Response in Neuronal Cells under Ischemic-like Conditions

O-GlcNAcylation is a nutrient-driven post-translational modification known as a metabolic sensor that links metabolism to cellular function. Recent evidences indicate that the activation of O-GlcNAc pathway is a potential pro-survival pathway and that acute enhancement of this response is conducive to the survival of cells and tissues. 2-(4-methoxyphenyl)ethyl-2-acetamido-2-deoxy-β-d-pyranoside (SalA-4g), is a salidroside analogue synthesized in our laboratory by chemical structure-modification, with a phenyl ring containing a para-methoxy group and a sugar ring consisting of N-acetylglucosamine. We have previously shown that SalA-4g elevates levels of protein O-GlcNAc and improves neuronal tolerance to ischemia. However, the specific target of SalA-4g regulating O-GlcNAcylation remains unknown. To address these questions, in this study, we have focused on mitochondrial network homeostasis mediated by O-GlcNAcylation in SalA-4g’s neuroprotection in primary cortical neurons under ischemic-like conditions. O-GlcNAc-modified mitochondria induced by SalA-4g demonstrated stronger neuroprotection under oxygen glucose deprivation and reoxygenation stress, including the improvement of mitochondrial homeostasis and bioenergy, and inhibition of mitochondrial apoptosis pathway. Blocking mitochondrial protein O-GlcNAcylation with OSMI-1 disrupted mitochondrial network homeostasis and antagonized the protective effects of SalA-4g. Collectively, these data demonstrate that mitochondrial homeostasis mediated by mitochondrial protein O-GlcNAcylation is critically involved in SalA-4g neuroprotection.

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Effects of O-GlcNAcylation on functional mitochondrial transfer from astrocytes

Journal of Cerebral Blood Flow & Metabolism ◽

10.1177/0271678x20969588 ◽

2020 ◽

pp. 0271678X2096958 ◽

Cited By ~ 1

Author(s):

Ji-Hyun Park ◽

Yoshihiko Nakamura ◽

Wenlu Li ◽

Gen Hamanaka ◽

Ken Arai ◽

...

Keyword(s):

Protein Modification ◽

Mitochondrial Protein ◽

Brefeldin A ◽

Glucose Deprivation ◽

Oxygen Glucose Deprivation ◽

Loss Of Function ◽

Post Translational Modification ◽

Mitochondrial Transfer ◽

The Central Nervous System ◽

Critical Process

Mitochondria may be transferred from cell to cell in the central nervous system and this process may help defend neurons against injury and disease. But how mitochondria maintain their functionality during the process of release into extracellular space remains unknown. Here, we report that mitochondrial protein O-GlcNAcylation is a critical process to support extracellular mitochondrial functionality. Activation of CD38-cADPR signaling in astrocytes robustly induced protein O-GlcNAcylation in mitochondria, while oxygen-glucose deprivation and reoxygenation showed transient and mild protein modification. Blocking the endoplasmic reticulum – Golgi trafficking with Brefeldin A or slc35B4 siRNA reduced O-GlcNAcylation, and resulted in the secretion of mitochondria with decreased membrane potential and mtDNA. Finally, loss-of-function studies verified that O-GlcNAc-modified mitochondria demonstrated higher levels of neuroprotection after astrocyte-to-neuron mitochondrial transfer. Collectively, these findings suggest that post-translational modification by O-GlcNAc may be required for supporting the functionality and neuroprotective properties of mitochondria released from astrocytes.

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Combination of mild hypothermia with neuroprotectants has greater neuroprotective effects during oxygen-glucose deprivation and reoxygenation-mediated neuronal injury

Scientific Reports ◽

10.1038/srep07091 ◽

2014 ◽

Vol 4 (1) ◽

Cited By ~ 27

Author(s):

Xiao-Ya Gao ◽

Jian-Ou Huang ◽

Ya-Fang Hu ◽

Yong Gu ◽

Shu-Zhen Zhu ◽

...

Keyword(s):

Cortical Neurons ◽

Mild Hypothermia ◽

Neuronal Damage ◽

Combined Treatment ◽

Neuronal Injury ◽

Glucose Deprivation ◽

Oxygen Glucose Deprivation ◽

Single Treatment ◽

Neuroprotective Effects ◽

Apoptosis Pathway

Abstract Co-treatment of neuroprotective reagents may improve the therapeutic efficacy of hypothermia in protecting neurons during ischemic stroke. This study aimed to find promising drugs that enhance the neuroprotective effect of mild hypothermia (MH). 26 candidate drugs were selected based on different targets. Primary cultured cortical neurons were exposed to oxygen-glucose deprivation and reoxygenation (OGD/R) to induce neuronal damage, followed by either single treatment (a drug or MH) or a combination of a drug and MH. Results showed that, compared with single treatment, combination of MH with brain derived neurotrophic factor, glibenclamide, dizocilpine, human urinary kallidinogenase or neuroglobin displayed higher proportion of neuronal cell viability. The latter three drugs also caused less apoptosis rate in combined treatment. Furthermore, co-treatment of those three drugs and MH decreased the level of reactive oxygen species (ROS) and intracellular calcium accumulation, as well as stabilized mitochondrial membrane potential (MMP), indicating the combined neuroprotective effects are probably via inhibiting mitochondrial apoptosis pathway. Taken together, the study suggests that combined treatment with hypothermia and certain neuroprotective reagents provide a better protection against OGD/R-induced neuronal injury.

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The protective effects of chitooligosaccharides against glucose deprivation-induced cell apoptosis in cultured cortical neurons through activation of PI3K/Akt and MEK/ERK1/2 pathways

Brain Research ◽

10.1016/j.brainres.2010.12.029 ◽

2011 ◽

Vol 1375 ◽

pp. 49-58 ◽

Cited By ~ 26

Author(s):

Yanyan Xu ◽

Qi Zhang ◽

Shu Yu ◽

Yumin Yang ◽

Fei Ding

Keyword(s):

Cell Apoptosis ◽

Cortical Neurons ◽

Glucose Deprivation ◽

Protective Effects ◽

Cultured Cortical Neurons

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Protective Effects and Mechanisms of Dendrobium nobile Lindl. Alkaloids on PC12 Cell Damage Induced by Aβ25-35

Behavioural Neurology ◽

10.1155/2021/9990375 ◽

2021 ◽

Vol 2021 ◽

pp. 1-11

Author(s):

Yuan Liu ◽

Tingting Pi ◽

Xiaohui Yang ◽

Jingshan Shi

Keyword(s):

Oxidative Stress ◽

Pc12 Cell ◽

Caspase 3 ◽

Cell Damage ◽

Protective Effects ◽

Related Protein ◽

Caspase 9 ◽

Apoptosis Pathway ◽

Mitochondrial Apoptosis Pathway ◽

Dendrobium Nobile Lindl

Background. Aβ deposition abnormally in the mitochondria can damage the mitochondrial respiratory chain and activate the mitochondrial-mediated apoptosis pathway, resulting in AD-like symptoms. Objective. To observe the protective effects of Dendrobium nobile Lindl. alkaloids (DNLA) on Aβ25-35-induced oxidative stress and apoptosis in PC12 cells explore its possible protective mechanisms. Methods. PC12 cells were treated with DNLA with different concentrations (0.035 mg/L, 0.3 mg/L, and 3.5 mg/L) for 6 h, followed by administration with Aβ25-35 (10 μM) for 24 h. MTT assay and flow cytometer observe the effect of DNLA on Aβ25-35-induced cytotoxicity and apoptosis of PC12 cell. Based on the mitochondrial apoptosis pathway to study the antiapoptotic effect of DNLA on this model and its relationship with oxidative stress, flow cytometer detected the level of reactive oxygen species (ROS), and ELISA kits were used to detect superoxide dismutase activity (SOD) and glutathione (GSH) content in cells. The JC-1 fluorescent staining observed the effect of DNLA on the mitochondrial membrane potential (MMP) with inverted immunofluorescence microscopy. Western blot was used to detect the levels of mitochondrial apoptosis pathway-related protein and its major downstream proteins Bax, Bcl-2, cleaved-caspase-9, and cleaved-caspase-3. Results. DNLA can significantly improve the viability and apoptosis rate of PC12 cell damage induced by Aβ25-35. It also can restore the reduced intracellular ROS content and MMP, while SOD activity and GSH content increase significantly. The expression of apoptosis-related protein Bax, cleaved-caspase-9, and cleaved-caspase-3 decreased when the Bcl-2 protein expression was significantly increased. Conclusion. These findings suggest that it can significantly inhibit the apoptosis of PC12 cell damage induced by Aβ25-35. The mechanism may reduce the level of cellular oxidative stress and thus inhibit the mitochondrial-mediated apoptosis pathway.

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Protective Effects of Chinese Propolis and Its Component, Chrysin, against Neuronal Cell Death via Inhibition of Mitochondrial Apoptosis Pathway in SH-SY5Y Cells

Journal of Agricultural and Food Chemistry ◽

10.1021/jf8014206 ◽

2008 ◽

Vol 56 (19) ◽

pp. 8944-8953 ◽

Cited By ~ 54

Author(s):

Hiroshi Izuta ◽

Masamitsu Shimazawa ◽

Shigemi Tazawa ◽

Yoko Araki ◽

Satoshi Mishima ◽

...

Keyword(s):

Cell Death ◽

Neuronal Cell Death ◽

Neuronal Cell ◽

Protective Effects ◽

Mitochondrial Apoptosis ◽

Apoptosis Pathway ◽

Mitochondrial Apoptosis Pathway ◽

Chinese Propolis

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Mitochondrial Carrier Homolog 2 Functionally Co-operates With BH3 Interacting-Domain Death Agonist in Promoting Ca2+-Induced Neuronal Injury

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.750100 ◽

2021 ◽

Vol 9 ◽

Author(s):

Beatrice D’Orsi ◽

Natalia Niewidok ◽

Heiko Düssmann ◽

Jochen H. M. Prehn

Keyword(s):

Cell Death ◽

Cortical Neurons ◽

Mitochondrial Permeability Transition ◽

Mitochondrial Protein ◽

Neuronal Cell ◽

Neuronal Injury ◽

Proteolytic Processing ◽

Apoptosis Pathway ◽

Mitochondrial Carrier ◽

Interacting Domain

The BH3 interacting-domain death agonist (BID) is a pro-apoptotic member of the Bcl-2 protein family. While proteolytic processing of BID links death receptor-induced apoptosis to the mitochondrial apoptosis pathway, we previously showed that full length BID also translocates to mitochondria during Ca2+-induced neuronal cell death. Moreover, mitochondrial carrier homolog 2 (MTCH2) was identified as a mitochondrial protein that interacts with BID during cell death. We started our studies by investigating the effect of Mtch2 silencing in a well-established model of Ca2+-induced mitochondrial permeability transition pore opening in non-neuronal HCT116 cells. We found that silencing of Mtch2 inhibited mitochondrial swelling and the associated decrease in mitochondrial energetics, suggesting a pro-death function for MTCH2 during Ca2+-induced injury. Next, we explored the role of BID and MTCH2 in mediating Ca2+-induced injury in primary cortical neurons triggered by prolonged activation of NMDA glutamate receptors. Analysis of intracellular Ca2+ transients, using time-lapse confocal microscopy, revealed that neurons lacking Bid showed markedly reduced Ca2+ levels during the NMDA excitation period. These Ca2+ transients were further decreased when Mtch2 was also silenced. Collectively, our data suggest that BID and MTCH2 functionally interact to promote Ca2+-induced neuronal injury.

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