Ubisol Coenzyme Q10 Promotes Mitochondrial Biogenesis in HT22 Cells Challenged by Glutamate

Abstract Background: Glutamate-induced excitotoxicity is a well-recognized cause of neuronal cell death and nutritional supplementation with Coenzyme Q10 (CoQ10) has previously been shown to have neuro-protective actions against it. The objective of this study was to determine whether the protective effect of CoQ10 against glutamate could be attributed to stimulating mitochondrial biogenesis. Results: The mouse hippocampal neuronal HT22 cells were incubated with glutamate with or without ubisol Q10 treatment. Significant deterioration of cells after glutamate exposure was observed under a light microscope and cell viability assay, along with a significant drop in clonogenic ability. Glutamate significantly decreased the mitochondrial biogenesis related protein levels of Akt, CREB, PGC-1α, and NRF2, and reduced mitochondrial biogenesis assessed by a mitochondrial biogenesis kit. Pretreatment with CoQ10 prevented the decreases of Akt, CREB, PGC-1α, and NRF2 and increased mitochondrial biogenesis. Conclusions: Taken together, these results describe a new mechanism of CoQ10-conveyed neuro-protection and indicate a central role for mitochondrial biogenesis in protecting against glutamate-induced excitotoxicity.

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BDMC protects AD in vitro via AMPK and SIRT1

Translational Neuroscience ◽

10.1515/tnsci-2020-0140 ◽

2020 ◽

Vol 11 (1) ◽

pp. 319-327

Author(s):

Chenlin Xu ◽

Zijian Xiao ◽

Heng Wu ◽

Guijuan Zhou ◽

Duanqun He ◽

...

Keyword(s):

Neurodegenerative Disorder ◽

Neuroprotective Effect ◽

Neuronal Cell Death ◽

Neuronal Cell ◽

Activity Measurement ◽

Therapeutic Approaches ◽

Cell Viability Assay ◽

Viability Assay ◽

Compound C

AbstractBackgroundAlzheimer’s disease (AD) is a common neurodegenerative disorder without any satisfactory therapeutic approaches. AD is mainly characterized by the deposition of β-amyloid protein (Aβ) and extensive neuronal cell death. Curcumin, with anti-oxidative stress (OS) and cell apoptosis properties, plays essential roles in AD. However, whether bisdemethoxycurcumin (BDMC), a derivative of curcumin, can exert a neuroprotective effect in AD remains to be elucidated.MethodsIn this study, SK-N-SH cells were used to establish an in vitro model to investigate the effects of BDMC on the Aβ1–42-induced neurotoxicity. SK-N-SH cells were pretreated with BDMC and with or without compound C and EX527 for 30 min after co-incubation with rotenone for 24 h. Subsequently, western blotting, cell viability assay and SOD and GSH activity measurement were performed.ResultsBDMC increased the cell survival, anti-OS ability, AMPK phosphorylation levels and SIRT1 in SK-N-SH cells treated with Aβ1–42. However, after treatment with compound C, an AMPK inhibitor, and EX527, an SIRT1inhibitor, the neuroprotective roles of BDMC on SK-N-SH cells treated with Aβ1–42 were inhibited.ConclusionThese results suggest that BDMC exerts a neuroprotective role on SK-N-SH cells in vitro via AMPK/SIRT1 signaling, laying the foundation for the application of BDMC in the treatment of neurodegenerative diseases related to AMPK/SIRT1 signaling.

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Insertion of the βGeo Promoter Trap into the Fem1c Gene of ROSA3 Mice

Molecular and Cellular Biology ◽

10.1128/mcb.24.9.3794-3803.2004 ◽

2004 ◽

Vol 24 (9) ◽

pp. 3794-3803 ◽

Cited By ~ 6

Author(s):

Cassandra L. Schlamp ◽

Andrew T. Thliveris ◽

Yan Li ◽

Louis P. Kohl ◽

Claudia Knop ◽

...

Keyword(s):

Cell Death ◽

Ganglion Cells ◽

Neuronal Cell Death ◽

Insertion Site ◽

Neuronal Cell ◽

Pyramidal Cells ◽

Coding Region ◽

Protein Levels ◽

Molecular Events ◽

Promoter Trap

ABSTRACT ROSA3 mice were developed by retroviral insertion of the βGeo gene trap vector. Adult ROSA3 mice exhibit widespread expression of the trap gene in epithelial cells found in most organs. In the central nervous system the highest expression of βGeo is found in CA1 pyramidal cells of the hippocampus, Purkinje cells of the cerebellum, and ganglion cells of the retina. Characterization of the genomic insertion site for βGeo in ROSA3 mice shows that the trap vector is located in the first intron of Fem1c, a gene homologous to the sex-determining gene fem-1 of Caenorhabditis elegans. Transcription of the Rosa3 allele (R3) yields a spliced message that includes the first exon of Fem1c and the βGeo coding region. Although normal processing of the Fem1c transcript is disrupted in homozygous Rosa3 (Fem1cR3/R3 ) mice, some tissues show low levels of a partially processed transcript containing exons 2 and 3. Since the entire coding region of Fem1c is located in these two exons, Fem1cR3/R3 mice may still be able to express a putative FEM1C protein. To this extent, Fem1cR3/R3 mice show no adverse effects in their sexual development or fertility or in the attenuation of neuronal cell death, another function that has been attributed to both fem-1 and a second mouse homolog, Fem1b. Examination of βGeo expression in ganglion cells after exposure to damaging stimuli indicates that protein levels are rapidly depleted prior to cell death, making the βGeo reporter gene a potentially useful marker to study early molecular events in damaged neurons.

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Ginsenoside Rb2 suppresses the glutamate-mediated oxidative stress and neuronal cell death in HT22 cells

Journal of Ginseng Research ◽

10.1016/j.jgr.2018.12.002 ◽

2019 ◽

Vol 43 (2) ◽

pp. 326-334 ◽

Cited By ~ 19

Author(s):

Dong Hoi Kim ◽

Dae Won Kim ◽

Bo Hyun Jung ◽

Jong Hun Lee ◽

Heesu Lee ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Death ◽

Neuronal Cell Death ◽

Neuronal Cell ◽

Ht22 Cells ◽

Ginsenoside Rb2

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Neuroprotective Effects of Tetrahydrocurcumin against Glutamate-Induced Oxidative Stress in Hippocampal HT22 Cells

Molecules ◽

10.3390/molecules25010144 ◽

2019 ◽

Vol 25 (1) ◽

pp. 144 ◽

Cited By ~ 4

Author(s):

Chang-Hyun Park ◽

Ji Hoon Song ◽

Su-Nam Kim ◽

Ji Hwan Lee ◽

Hae-Jeung Lee ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Death ◽

Protein Kinases ◽

Apoptotic Cell ◽

Curcuma Longa ◽

Neuronal Cell Death ◽

Neuronal Cell ◽

Ht22 Cells ◽

Mitogen Activated Protein Kinases ◽

Mitogen Activated Protein

In the central nervous system, glutamate is a major excitable neurotransmitter responsible for many cellular functions. However, excessive levels of glutamate induce neuronal cell death via oxidative stress during acute brain injuries as well as chronic neurodegenerative diseases. The present study was conducted to examine the effect of tetrahydrocurcumin (THC), a major secondary metabolite of curcumin, and its possible mechanism against glutamate-induced cell death. We prepared THC using curcumin isolated from Curcuma longa (turmeric) and demonstrated the protective effect of THC against glutamate-induced oxidative stress in HT22 cells. THC abrogated glutamate-induced HT22 cell death and showed a strong antioxidant effect. THC also significantly reduced intracellular calcium ion increased by glutamate. Additionally, THC significantly reduced the accumulation of intracellular oxidative stress induced by glutamate. Furthermore, THC significantly diminished apoptotic cell death indicated by annexin V-positive in HT22 cells. Western blot analysis indicated that the phosphorylation of mitogen-activated protein kinases including c-Jun N-terminal kinase, extracellular signal-related kinases 1/2, and p38 by glutamate was significantly diminished by treatment with THC. In conclusion, THC is a potent neuroprotectant against glutamate-induced neuronal cell death by inhibiting the accumulation of oxidative stress and phosphorylation of mitogen-activated protein kinases.

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Peroxiredoxin 5 Inhibits Glutamate-Induced Neuronal Cell Death through the Regulation of Calcineurin-Dependent Mitochondrial Dynamics in HT22 Cells

Molecular and Cellular Biology ◽

10.1128/mcb.00148-19 ◽

2019 ◽

Vol 39 (20) ◽

Cited By ~ 6

Author(s):

Mi Hye Kim ◽

Hong Jun Lee ◽

Sang-Rae Lee ◽

Hyun-Shik Lee ◽

Jae-Won Huh ◽

...

Keyword(s):

Cell Death ◽

Neurodegenerative Diseases ◽

Mitochondrial Dynamics ◽

Mitochondrial Fission ◽

Neuronal Cell Death ◽

Neuronal Cell ◽

Glutamate Toxicity ◽

Ht22 Cells ◽

Peroxidase Enzyme ◽

The Central Nervous System

ABSTRACT Glutamate is an essential neurotransmitter in the central nervous system (CNS). However, high glutamate concentrations can lead to neurodegenerative diseases. A hallmark of glutamate toxicity is high levels of reactive oxygen species (ROS), which can trigger Ca2+ influx and dynamin-related protein 1 (Drp1)-mediated mitochondrial fission. Peroxiredoxin 5 (Prx5) is a well-known cysteine-dependent peroxidase enzyme. However, the precise effects of Prx5 on glutamate toxicity are still unclear. In this study, we investigated the role of Prx5 in glutamate-induced neuronal cell death. We found that glutamate treatment induces endogenous Prx5 expression and Ca2+/calcineurin-dependent dephosphorylation of Drp1, resulting in mitochondrial fission and neuronal cell death. Our results indicate that Prx5 inhibits glutamate-induced mitochondrial fission through the regulation of Ca2+/calcineurin-dependent dephosphorylation of Drp1, and it does so by scavenging cytosolic and mitochondrial ROS. Therefore, we suggest that Ca2+/calcineurin-dependent mitochondrial dynamics are deeply associated with glutamate-induced neurotoxicity. Consequently, Prx5 may be used as a potential agent for developing therapies against glutamate-induced neurotoxicity and neurodegenerative diseases where it plays a key role.

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Cerebral Ischemia Causes Dysregulation of Synaptic Adhesion in Mouse Synaptosomes

Journal of Cerebral Blood Flow & Metabolism ◽

10.1038/sj.jcbfm.9600510 ◽

2007 ◽

Vol 28 (1) ◽

pp. 99-110 ◽

Cited By ~ 15

Author(s):

Willard J Costain ◽

Ingrid Rasquinha ◽

Jagdeep K Sandhu ◽

Peter Rippstein ◽

Bogdan Zurakowski ◽

...

Keyword(s):

Cell Death ◽

Cerebral Ischemia ◽

Adhesion Molecules ◽

Neuronal Cell Death ◽

Neuronal Cell ◽

Artery Occlusion ◽

Synaptic Proteins ◽

Mrna Levels ◽

Neuronal Nuclei ◽

Protein Levels

Synaptic pathology is observed during hypoxic events in the central nervous system in the form of altered dendrite structure and conductance changes. These alterations are rapidly reversible, on the return of normoxia, but are thought to initiate subsequent neuronal cell death. To characterize the effects of hypoxia on regulators of synaptic stability, we examined the temporal expression of cell adhesion molecules (CAMs) in synaptosomes after transient middle cerebral artery occlusion (MCAO) in mice. We focused on events preceding the onset of ischemic neuronal cell death (< 48 h). Synaptosome preparations were enriched in synaptically localized proteins and were free of endoplasmic reticulum and nuclear contamination. Electron microscopy showed that the synaptosome preparation was enriched in spheres (≈650 nm in diameter) containing secretory vesicles and postsynaptic densities. Forebrain mRNA levels of synaptically located CAMs was unaffected at 3 h after MCAO. This is contrasted by the observation of consistent downregulation of synaptic CAMs at 20 h after MCAO. Examination of synaptosomal CAM protein content indicated that certain adhesion molecules were decreased as early as 3 h after MCAO. For comparison, synaptosomal Agrn protein levels were unaffected by cerebral ischemia. Furthermore, a marked increase in the levels of p-Ctnnb1 in ischemic synaptosomes was observed. p-Ctnnb1 was detected in hippocampal fiber tracts and in cornu ammonis 1 neuronal nuclei. These results indicate that ischemia induces a dysregulation of a subset of synaptic proteins that are important regulators of synaptic plasticity before the onset of ischemic neuronal cell death.

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Peroxiredoxin II Maintains the Mitochondrial Membrane Potential against Alcohol-Induced Apoptosis in HT22 Cells

Antioxidants ◽

10.3390/antiox9010001 ◽

2019 ◽

Vol 9 (1) ◽

pp. 1

Author(s):

Mei-Hua Jin ◽

Jia-Bin Yu ◽

Hu-Nan Sun ◽

Ying-Hua Jin ◽

Gui-Nan Shen ◽

...

Keyword(s):

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Mitochondrial Membrane ◽

Neuronal Cell Death ◽

Neuronal Cell ◽

Ht22 Cells ◽

Apoptotic Protein ◽

Intracellular Ros ◽

Induced Apoptosis ◽

The Brain

Excessive alcohol intake can significantly reduce cognitive function and cause irreversible learning and memory disorders. The brain is particularly vulnerable to alcohol-induced ROS damage; the hippocampus is one of the most sensitive areas of the brain for alcohol neurotoxicity. In the present study, we observed significant increasing of intracellular ROS accumulations in Peroxiredoxin II (Prx II) knockdown HT22 cells, which were induced by alcohol treatments. We also found that the level of ROS in mitochondrial was also increased, resulting in a decrease in the mitochondrial membrane potential. The phosphorylation of GSK3β (Ser9) and anti-apoptotic protein Bcl2 expression levels were significantly downregulated in Prx II knockdown HT22 cells, which suggests that Prx II knockdown HT22 cells were more susceptible to alcohol-induced apoptosis. Scavenging the alcohol-induced ROS with NAC significantly decreased the intracellular ROS levels, as well as the phosphorylation level of GSK3β in Prx II knockdown HT22 cells. Moreover, NAC treatment also dramatically restored the mitochondrial membrane potential and the cellular apoptosis in Prx II knockdown HT22 cells. Our findings suggest that Prx II plays a crucial role in alcohol-induced neuronal cell apoptosis by regulating the cellular ROS levels, especially through regulating the ROS-dependent mitochondrial membrane potential. Consequently, Prx II may be a therapeutic target molecule for alcohol-induced neuronal cell death, which is closely related to ROS-dependent mitochondria dysfunction.

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Efficient Delivery of Therapeutic siRNA with Nanoparticles Induces Apoptosis in Prostate Cancer Cells

Journal of Nanomaterials ◽

10.1155/2018/4719790 ◽

2018 ◽

Vol 2018 ◽

pp. 1-10 ◽

Cited By ~ 4

Author(s):

Xupeng Mu ◽

Haibin Lu ◽

Lianlian Fan ◽

Shaohua Yan ◽

Kebang Hu

Keyword(s):

Sirna Delivery ◽

Mitochondrial Pathway ◽

Gene Knockdown ◽

Hybrid Nanoparticles ◽

Cell Viability Assay ◽

Protein Levels ◽

Viability Assay ◽

Efficient Delivery ◽

Du145 Cells

Gene silencing using small interfering RNA (siRNA) has shown significant potential in the treatment of cancer. Herein, we developed the lipid-polymer hybrid nanoparticles (PEG-LP/siRNA NPs) for siRNA delivery. The cell viability assay indicated that PEG-LP/siRNA NPs had negligible cell cytotoxicity. The cellular uptake efficiency of PEG-LP/siRNA NPs measured by flow cytometry was up to 94.4%. Importantly, in vitro gene knockdown experiments demonstrated that PEG-LP/siJnk-1 NPs could significantly downregulate the expression of Jnk-1 at both the mRNA and protein levels in DU145 cells. Gene knockdown of Jnk-1 could activate apoptosis in part by the mitochondrial pathway in DU145 cells. Moreover, the PEG-LP/siJnk-1 NPs could inhibit tumor growth in a DU145 xenograft murine model, suggesting its therapeutic promise in cancer therapy.

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Dual Targeting by Inhibition of Phosphoinositide-3-Kinase and Mammalian Target of Rapamycin Attenuates the Neuroinflammatory Responses in Murine Hippocampal Cells and Seizures in C57BL/6 Mice

Frontiers in Immunology ◽

10.3389/fimmu.2021.739452 ◽

2021 ◽

Vol 12 ◽

Author(s):

Preeti Vyas ◽

Rajkumar Tulsawani ◽

Divya Vohora

Keyword(s):

Cell Signaling ◽

Day Treatment ◽

Neuronal Cell Death ◽

Neuronal Cell ◽

Neuronal Loss ◽

Ht22 Cells ◽

Seizure Severity ◽

Combination Model

Emerging evidence suggests the association of seizures and inflammation; however, underlying cell signaling mechanisms are still not fully understood. Overactivation of phosphoinositide-3-kinases is associated with both neuroinflammation and seizures. Herein, we speculate the PI3K/Akt/mTOR pathway as a promising therapeutic target for neuroinflammation-mediated seizures and associated neurodegeneration. Firstly, we cultured HT22 cells for detection of the downstream cell signaling events activated in a lipopolysaccharide (LPS)-primed pilocarpine (PILO) model. We then evaluated the effects of 7-day treatment of buparlisib (PI3K inhibitor, 25 mg/kg p.o.), dactolisib (PI3K/mTOR inhibitor, 25 mg/kg p.o.), and rapamycin (mTORC1 inhibitor, 10 mg/kg p.o.) in an LPS-primed PILO model of seizures in C57BL/6 mice. LPS priming resulted in enhanced seizure severity and reduced latency. Buparlisib and dactolisib, but not rapamycin, prolonged latency to seizures and reduced neuronal loss, while all drugs attenuated seizure severity. Buparlisib and dactolisib further reduced cellular redox, mitochondrial membrane potential, cleaved caspase-3 and p53, nuclear integrity, and attenuated NF-κB, IL-1β, IL-6, TNF-α, and TGF-β1 and TGF-β2 signaling both in vitro and in vivo post-PILO and LPS+PILO inductions; however, rapamycin mitigated the same only in the PILO model. Both drugs protected against neuronal cell death demonstrating the contribution of this pathway in the seizure-induced neuronal pyknosis; however, rapamycin showed resistance in a combination model. Furthermore, LPS and PILO exposure enhanced pAkt/Akt and phospho-p70S6/total-p70S6 kinase activity, while buparlisib and dactolisib, but not rapamycin, could reduce it in a combination model. Partial rapamycin resistance was observed possibly due to the reactivation of the pathway by a functionally different complex of mTOR, i.e., mTORC2. Our study substantiated the plausible involvement of PI3K-mediated apoptotic and inflammatory pathways in LPS-primed PILO-induced seizures and provides evidence that its modulation constitutes an anti-inflammatory mechanism by which seizure inhibitory effects are observed. We showed dual inhibition by dactolisib as a promising approach. Targeting this pathway at two nodes at a time may provide new avenues for antiseizure therapies.

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Calcium-dependent serine-threonine phosphatase, calcineurin inactivation mediated by baicalein attenuates prion protein-mediated neuronal cell damage

10.21203/rs.3.rs-18414/v6 ◽

2021 ◽

Author(s):

Jeong-Min Hong ◽

Ji-Hong Moon ◽

Young Min Oh ◽

Sang-Youel Park

Keyword(s):

Cell Death ◽

Prion Protein ◽

Cell Damage ◽

Prion Diseases ◽

Neuronal Cell Death ◽

Neuronal Cell ◽

Therapeutic Drug ◽

Autophagic Flux ◽

Calcium Dependent ◽

Protein Levels

Abstract Background: Prion diseases are a group of unvaryingly fatal neurodegenerative disorders characterized by neuronal cell death. Calcineurin and autophagy mediate prion-induced neurodegeneration, suggesting that inhibition of calcineurin and autophagy could be a target for therapy. Baicalein has been reported to exert neuroprotective effects against calcium-dependent neuronal cell death. Results: In the present study, we investigated whether baicalein attenuates prion peptide-mediated neurotoxicity and reduces calcineurin. We found that baicalein treatment inhibits prion protein-induced apoptosis. Baicalein inhibited calcium up-regulation and protected the cells against prion peptide‑induced neuron cell death by calcineurin inactivation. Furthermore, baicalein increased p62 protein levels and decrease LC3-II protein levels indicating autophagic flux inhibition and baicalein inhibited prion protein-induced neurotoxicity through autophagy flux inhibition. Conclusions: Taken together, this study demonstrated that baicalein attenuated prion peptide-induced neurotoxicity via calcineurin inactivation and autophagic flux reduction, and also suggest that baicalein may be an effective therapeutic drug against neurodegenerative diseases, including prion diseases.

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