Neuroprotective Effects of Magnesium L-threonate in a Hypoxic Zebrafish Model

Abstract Background: Hypoxia inhibits the uptake of glutamate (a major neurotransmitter in the brain closely related to cognitive function) into brain cells, and the initial response of cells to cortical hypoxia depends on glutamate. Previous studies have suggested that magnesium may have protective effects against hypoxic injuries. In particular, magnesium L-threonate (MgT) may increase magnesium ion concentrations in the brain better than MgSO4 and improve cognitive function. Methods: We evaluated cell viability under hypoxic conditions in the MgT- and MgSO4-treated human SH-SY5Y neurons, in vivo behavior using the T-maze test following hypoxia in MgT-treated zebrafish, activity of brain mitochondrial dehydrogenase by 2,3,5-triphenyltetrazolium chloride (TTC) staining, and protein expression of the excitatory amino acid transporter (EAAT) 4 glutamate transporter by western blotting. Results: Among the groups treated with hypoxia, cell viability significantly increased when pre-treated with 1 or 10 mM MgT (p = 0.009 and 0.026, respectively) Despite hypoxic insult, MgT-treated zebrafish showed preferences for the red compartment (p= 0.025 for distance and p= 0.007 for frequency of entries), suggesting memory preservation. TTC staining showed reduced cerebral infarction and preserved absorbance in the MgT-treated zebrafish brain after hypoxia (p=0.010 compared to the hypoxia group). In addition, western blot showed upregulation of EAAT4 protein in the MgT treated group. Conclusions: Pre-treatment with MgT attenuated cell death and cerebral infarction due to hypoxia and protected cognitive function in zebrafish. In addition, MgT appeared to modulate expression of the glutamate transporter, EAAT4.

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Neuroprotective Effects of Magnesium L-threonate in a Hypoxic Zebrafish Model

10.21203/rs.2.19975/v3 ◽

2020 ◽

Author(s):

Young Sung Kim ◽

Young Ju Won ◽

Byung Gun Lim ◽

Too Jae Min ◽

Yeon-Hwa Kim ◽

...

Keyword(s):

Cognitive Function ◽

Cerebral Infarction ◽

Cell Viability ◽

Excitatory Amino Acid ◽

Glutamate Transporter ◽

Protective Effects ◽

Triphenyltetrazolium Chloride ◽

Zebrafish Model ◽

Neuroprotective Effects ◽

The Brain

Abstract Background: Hypoxia inhibits the uptake of glutamate (a major neurotransmitter in the brain closely related to cognitive function) into brain cells, and the initial response of cells to cortical hypoxia depends on glutamate. Previous studies have suggested that magnesium may have protective effects against hypoxic injuries. In particular, magnesium L-threonate (MgT) may increase magnesium ion concentrations in the brain better than MgSO4 and improve cognitive function. Methods: We evaluated cell viability under hypoxic conditions in the MgT- and MgSO4-treated human SH-SY5Y neurons, in vivo behavior using the T-maze test following hypoxia in MgT-treated zebrafish, activity of brain mitochondrial dehydrogenase by 2,3,5-triphenyltetrazolium chloride (TTC) staining, and protein expression of the excitatory amino acid transporter (EAAT) 4 glutamate transporter by western blotting.Results: Among the groups treated with hypoxia, cell viability significantly increased when pre-treated with 1 or 10 mM MgT (p=0.009 and 0.026, respectively). Despite hypoxic insult, MgT-treated zebrafish showed preferences for the red compartment (p=0.025 for distance and p=0.007 for frequency of entries), suggesting memory preservation. TTC staining showed reduced cerebral infarction and preserved absorbance in the MgT-treated zebrafish brain after hypoxia (p=0.010 compared to the hypoxia group). In addition, western blot showed upregulation of EAAT4 protein in the MgT treated group.Conclusions: Pre-treatment with MgT attenuated cell death and cerebral infarction due to hypoxia and protected cognitive function in zebrafish. In addition, MgT appeared to modulate expression of the glutamate transporter, EAAT4.

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Neuroprotective Effects of Magnesium L-threonate in a Hypoxic Zebrafish Model

10.21203/rs.2.19975/v4 ◽

2020 ◽

Author(s):

Young Sung Kim ◽

Young Ju Won ◽

Byung Gun Lim ◽

Too Jae Min ◽

Yeon-Hwa Kim ◽

...

Keyword(s):

Cognitive Function ◽

Cerebral Infarction ◽

Cell Viability ◽

Excitatory Amino Acid ◽

Glutamate Transporter ◽

Protective Effects ◽

Triphenyltetrazolium Chloride ◽

Zebrafish Model ◽

Neuroprotective Effects ◽

The Brain

Abstract Background: Hypoxia inhibits the uptake of glutamate (a major neurotransmitter in the brain closely related to cognitive function) into brain cells, and the initial response of cells to cortical hypoxia depends on glutamate. Previous studies have suggested that magnesium may have protective effects against hypoxic injuries. In particular, magnesium L-threonate (MgT) may increase magnesium ion concentrations in the brain better than MgSO 4 and improve cognitive function. Methods: We evaluated cell viability under hypoxic conditions in the MgT- and MgSO 4 -treated human SH-SY5Y neurons, in vivo behavior using the T-maze test following hypoxia in MgT-treated zebrafish, activity of brain mitochondrial dehydrogenase by 2,3,5-triphenyltetrazolium chloride (TTC) staining, and protein expression of the excitatory amino acid transporter (EAAT) 4 glutamate transporter by western blotting. Results : Among the groups treated with hypoxia, cell viability significantly increased when pre-treated with 1 or 10 mM MgT (p = 0.009 and 0.026, respectively). Despite hypoxic insult, MgT-treated zebrafish showed preferences for the red compartment (p= 0.025 for distance and p= 0.007 for frequency of entries), suggesting memory preservation. TTC staining showed reduced cerebral infarction and preserved absorbance in the MgT-treated zebrafish brain after hypoxia (p=0.010 compared to the hypoxia group). In addition, western blot showed upregulation of EAAT4 protein in the MgT treated group. Conclusions: Pre-treatment with MgT attenuated cell death and cerebral infarction due to hypoxia and protected cognitive function in zebrafish. In addition, MgT appeared to modulate expression of the glutamate transporter, EAAT4.

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Neuroprotective Effects of Magnesium L-threonate in a Hypoxic Zebrafish Model

10.21203/rs.2.19975/v2 ◽

2020 ◽

Author(s):

Young Sung Kim ◽

Young Ju Won ◽

Byung Gun Lim ◽

Too Jae Min ◽

Yeon-Hwa Kim ◽

...

Keyword(s):

Cognitive Function ◽

Cerebral Infarction ◽

Cell Viability ◽

Excitatory Amino Acid ◽

Glutamate Transporter ◽

Protective Effects ◽

Triphenyltetrazolium Chloride ◽

Zebrafish Model ◽

Neuroprotective Effects ◽

The Brain

Abstract Background: Hypoxia inhibits the uptake of glutamate (a major neurotransmitter in the brain closely related to cognitive function) into brain cells, and the initial response of cells to cortical hypoxia depends on glutamate. Previous studies have suggested that magnesium may have protective effects against hypoxic injuries. In particular, magnesium L-threonate (MgT) may increase magnesium ion concentrations in the brain better than MgSO 4 and improve cognitive function.Methods: We evaluated cell viability under hypoxic conditions in the MgT- and MgSO 4 -treated human SH-SY5Y neurons, in vivo behavior using the T-maze test following hypoxia in MgT-treated zebrafish, activity of brain mitochondrial dehydrogenase by 2,3,5-triphenyltetrazolium chloride (TTC) staining, and protein expression of the excitatory amino acid transporter (EAAT) 4 glutamate transporter by western blotting.Results : Among the groups treated with hypoxia, cell viability significantly increased when pre-treated with 1 or 10 mM MgT (p = 0.009 and 0.026, respectively) Despite hypoxic insult, MgT-treated zebrafish showed preferences for the red compartment (p= 0.025 for distance and p= 0.007 for frequency of entries), suggesting memory preservation. TTC staining showed reduced cerebral infarction and preserved absorbance in the MgT-treated zebrafish brain after hypoxia (p=0.010 compared to the hypoxia group). In addition, western blot showed upregulation of EAAT4 protein in the MgT treated group.Conclusions: Pre-treatment with MgT attenuated cell death and cerebral infarction due to hypoxia and protected cognitive function in zebrafish. In addition, MgT appeared to modulate expression of the glutamate transporter, EAAT4.

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Glutathione in the Brain

International Journal of Molecular Sciences ◽

10.3390/ijms22095010 ◽

2021 ◽

Vol 22 (9) ◽

pp. 5010

Author(s):

Koji Aoyama

Keyword(s):

Neurodegenerative Diseases ◽

Regulatory Mechanism ◽

Excitatory Amino Acid ◽

Redox Homeostasis ◽

Disease Onset ◽

Glutamate Transporter ◽

Antioxidant Defense System ◽

Common Finding ◽

Potential Therapeutic Application ◽

The Brain

Glutathione (GSH) is the most abundant non-protein thiol, and plays crucial roles in the antioxidant defense system and the maintenance of redox homeostasis in neurons. GSH depletion in the brain is a common finding in patients with neurodegenerative diseases, such as Alzheimer’s disease and Parkinson’s disease, and can cause neurodegeneration prior to disease onset. Excitatory amino acid carrier 1 (EAAC1), a sodium-dependent glutamate/cysteine transporter that is selectively present in neurons, plays a central role in the regulation of neuronal GSH production. The expression of EAAC1 is posttranslationally controlled by the glutamate transporter-associated protein 3–18 (GTRAP3-18) or miR-96-5p in neurons. The regulatory mechanism of neuronal GSH production mediated by EAAC1 may be a new target in therapeutic strategies for these neurodegenerative diseases. This review describes the regulatory mechanism of neuronal GSH production and its potential therapeutic application in the treatment of neurodegenerative diseases.

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Evaluation of the Anti-apoptotic and Anti-cytotoxic Effect of Epicatechin Gallate and Edaravone on SH-SY5Y Neuroblastoma Cells

Basic and Clinical Neuroscience Journal ◽

10.32598/bcn.9.10.1159.2 ◽

2019 ◽

pp. 619-630

Author(s):

Mohammad Shokrzadeh ◽

Hashem Javanmard ◽

Golpar Golmohammad Zadeh ◽

Hossein Asgarian Omran ◽

Mona Modanlou ◽

...

Keyword(s):

Synergistic Effect ◽

Cell Viability ◽

Mtt Assay ◽

Neuroprotective Effect ◽

Neuroblastoma Cells ◽

Protective Effects ◽

Older Individuals ◽

Epicatechin Gallate ◽

Neuroprotective Effects ◽

Late Apoptosis

Introduction: Parkinson disease (PD) is the second most common neurodegenerative disease affecting older individuals with signs of motor disability and cognitive impairment. Epicatechin (EC) and edaravone have neuroprotective effects most probably due to their antioxidant activity; however, a limited number of studies have considered their role in PD. This research aimed at investigating the neuroprotective effect of EC and edaravone in a neurotoxin-induced model of PD. Methods: An in vitro model of PD was made by subjecting SH-SY5Y neuroblastoma cells to neurotoxin: 6-hydroxydopamine (6-OHDA) 100 µM/well. The cytoprotective effect of EC and edaravone in five concentrations on cell viability was tested using the MTT assay. The apoptotic assay was done by annexin V and propidium iodide method using flow cytometry. Results: According to the MTT assay analysis, EC and edaravone had protective effects against 6-OH DA-induced cytotoxicity in SH-SY5Y neuroblastoma cells that were much more significant for edaravone and also a relative synergistic effect between EC and edaravone was observed. The apoptotic analysis showed that edaravone alone could decrease early and late apoptosis, whereas EC diminished early apoptosis, but enhanced late apoptosis and necrosis. Besides, co-treatment of edaravone and EC had a synergistic effect on decreasing apoptosis and increasing cell viability. Conclusion: The protective effect of edaravone on apoptosis and cytotoxicity was demonstrated clearly and EC had a synergistic effect with edaravone.

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Hesperetin protects SH-SY5Y cells against 6- hydroxydopamine-induced neurotoxicity via activation of NRF2/ARE signaling pathways

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v19i6.12 ◽

2020 ◽

Vol 19 (6) ◽

pp. 1197-1201 ◽

Cited By ~ 1

Author(s):

Jing Li ◽

Yue Liu ◽

Li Wang ◽

Zhaowei Gu ◽

Zhigang Huan ◽

...

Keyword(s):

Oxidative Stress ◽

Parkinson’S Disease ◽

Parkinson's Disease ◽

Cell Viability ◽

Heme Oxygenase 1 ◽

Protective Effects ◽

Neuroprotective Effects ◽

Ldh Release ◽

6 Hydroxydopamine

Purpose: To investigation the protective effects of hesperetin against 6-hydroxydopamine (6-OHDA)- induced neurotoxicity. Methods: SH-SY5Y cells were incubated with 6-OHDA to create an in vitro model of neurotoxicity. This model was used to test the neuroprotective effects of hesperetin. Cell viability was assessed by MTT and lactate dehydrogenase (LDH) release assays. Flow cytometry and western blot were used to quantify apoptosis. Oxidative stress was evaluated by determining intracellular glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), and reactive oxygen species (ROS). Results: In SH-SY5Y cells, treatment with 6-OHDA decreased cell viability and promoted LDH release. However, exogenous hesperetin protected against 6-OHDA-mediated toxicity. Similarly, although incubation with 6-OHDA induced apoptosis and increased cleaved caspase-3 and -9 levels, treatment with hesperetin protected against these effects. Treatment with 6-OHDA also led to significant oxidative stress, as indicated by reduced GSH and SOD levels and increased MDA and ROS levels in SH-SY5Y cells. However, these changes were reversed by pre-treatment with hesperetin. Of interest, hesperetin led to changes in 6-OHDA-induced expression of NRF2, heme oxygenase-1 (HO-1), glutamate-cysteine ligase (GCL) catalytic subunit (GCLC), and GCL modulatory (GCLM). Conclusion: Hesperetin protects against cell toxicity, apoptosis, and oxidative stress via activation of NRF2 pathway in a 6-OHDA-induced model of neurotoxicity. Future studies should investigate the use of hesperetin as a potential therapeutic approach for prevention or management of Parkinson’s disease. Keywords: Hesperetin, 6-OHDA, Neurotoxicity, NRF2, Parkinson’s disease

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Effect of growth hormone and melatonin on the brain: from molecular mechanisms to structural changes

Hormone Molecular Biology and Clinical Investigation ◽

10.1515/hmbci.2011.115 ◽

2011 ◽

Vol 7 (2) ◽

Author(s):

Roman A. Kireev ◽

Sara Cuesta ◽

Elena Vara ◽

Jesus A.F. Tresguerres

Keyword(s):

Growth Hormone ◽

Structural Changes ◽

Molecular Mechanisms ◽

Economic Consequences ◽

Aging Brain ◽

Protective Effects ◽

Negative Effects ◽

Gh Treatment ◽

Neuroprotective Effects ◽

The Brain

AbstractAging of the brain causes important reductions in quality of life and has wide socio-economic consequences. An increase in oxidative stress, and the associated inflammation and apoptosis, could be responsible for the pathogenesis of aging associated brain lesions. Melatonin has neuroprotective effects, by limiting the negative effects of oxygen and nitrogen free radicals. Growth hormone (GH) might exert additional neuro-protective and or neurogenic effects on the brain. The molecular mechanisms of the protective effects of GH and melatonin on the aging brain have been investigated in young and old Wistar rats. A reduction in the total number of neurons in the hilus of the dentate gyrus was evident at 24 months of age and was associated with a significant increase in inflammation markers as well as in pro-apoptotic parameters, confirming the role of apoptosis in its reduction. Melatonin treatment was able to enhance neurogenesis in old rats without modification of the total number of neurons, whereas GH treatment increased the total number of neurons without enhancing neurogenesis. Both GH and melatonin were able to reduce inflammation and apoptosis in the hippocampus. In conclusion, neuroprotective effects demonstrated by GH and melatonin in the hippocampus were exerted by decreasing inflammation and apoptosis.

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Hexarelin Modulation of MAPK and PI3K/Akt Pathways in Neuro-2A Cells Inhibits Hydrogen Peroxide—Induced Apoptotic Toxicity

Pharmaceuticals ◽

10.3390/ph14050444 ◽

2021 ◽

Vol 14 (5) ◽

pp. 444

Author(s):

Ramona Meanti ◽

Laura Rizzi ◽

Elena Bresciani ◽

Laura Molteni ◽

Vittorio Locatelli ◽

...

Keyword(s):

Hydrogen Peroxide ◽

Cell Viability ◽

Caspase 3 ◽

Mrna Levels ◽

Protective Effects ◽

Neuroprotective Effects ◽

Caspase 7 ◽

Induced Apoptosis

Hexarelin, a synthetic hexapeptide, exerts cyto-protective effects at the mitochondrial level in cardiac and skeletal muscles, both in vitro and in vivo, may also have important neuroprotective bioactivities. This study examined the inhibitory effects of hexarelin on hydrogen peroxide (H2O2)-induced apoptosis in Neuro-2A cells. Neuro-2A cells were treated for 24 h with various concentrations of H2O2 or with the combination of H2O2 and hexarelin following which cell viability and nitrite (NO2−) release were measured. Cell morphology was also documented throughout and changes arising were quantified using Image J skeleton and fractal analysis procedures. Apoptotic responses were evaluated by Real-Time PCR (caspase-3, caspase-7, Bax, and Bcl-2 mRNA levels) and Western Blot (cleaved caspase-3, cleaved caspase-7, MAPK, and Akt). Our results indicate that hexarelin effectively antagonized H2O2-induced damage to Neuro-2A cells thereby (i) improving cell viability, (ii) reducing NO2− release and (iii) restoring normal morphologies. Hexarelin treatment also reduced mRNA levels of caspase-3 and its activation, and modulated mRNA levels of the BCL-2 family. Moreover, hexarelin inhibited MAPKs phosphorylation and increased p-Akt protein expression. In conclusion, our results demonstrate neuroprotective and anti-apoptotic effects of hexarelin, suggesting that new analogues could be developed for their neuroprotective effects.

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Comparison of the protective effects of Scrophularia striata extract with vitamin E on cognitive function, anxiety and pain sensitivity in diazinon-exposed male rats

Physiology and Pharmacology ◽

10.32598/ppj.25.1.30 ◽

2020 ◽

Vol 25 (1) ◽

pp. 47-56

Author(s):

Ali Abedi ◽

◽

Khayam Bamdad ◽

Hilda Yaghoubi Shahir ◽

Raziyeh Dehghany ◽

...

Keyword(s):

Body Weight ◽

Vitamin E ◽

Cognitive Function ◽

Pain Sensitivity ◽

Male Rats ◽

Thermal Stimulus ◽

Protective Effects ◽

Hot Plate ◽

Neuroprotective Effects ◽

Scrophularia Striata

Introduction: Scrophularia striata is used in traditional medicine to treat various disorders and has neuroprotective effects. There are no studies about the effects of S. striata on cognitive functions in diazinon (DZN)- exposed rats. According to the results of previous studies, vitamin E (Vit. E) can also act as a protective agent against cognitive impairments. Therefore, the present study was designed to compare the effects of Vit. E and S. striata on DZN-induced behavioral impairments in male rats. Methods: Neuroprotective effects of S. striata (30mg/kg, 5 days/week for 8 weeks) and Vit. E (200mg/kg, 5 days/week for 8 weeks, IP) were assessed through changes in memory, anxiety-like behaviors and pain threshold following DZN exposure. Open field, shuttle box and hot plate were used to examine anxiety-like behaviors, passive avoidance learning and memory as well as pain sensitivity, respectively. Results: Our findings indicated that exposure to DZN caused a significant decrease in memory retention and an increase in anxiety-like behaviors. S. striata and Vit. E administration compensated memory and emotional impairments induced by DZN. As well as, S. striata alone decreased reaction time against thermal stimulus in the hot plate test. The findings of the present study also indicated that exposure to DZN significantly decreased body weight, while S. striata and Vit. E consumption restored it. Conclusion: Results of our study indicated the protective effects of S. striata consumption like Vit. E against DZN-induced disruptions in anxiety, cognitive function and body weight loss.

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The Neuroprotective Effects of Astragaloside IV against H2O2-Induced Damage in SH-SY5Y Cells are Associated with Synaptic Plasticity

Journal of Chemistry ◽

10.1155/2020/5343619 ◽

2020 ◽

Vol 2020 ◽

pp. 1-7

Author(s):

Zurong Song ◽

Ali Tao

Keyword(s):

Synaptic Plasticity ◽

Western Blot ◽

Cell Viability ◽

Cell Damage ◽

Neuroblastoma Cell ◽

Human Neuroblastoma Cell ◽

Protective Effects ◽

Astragaloside Iv ◽

Neuroprotective Effects ◽

Human Neuroblastoma

The aim of this study was to investigate whether the neuroprotective effects of astragaloside IV (AS-IV) against hydrogen peroxide (H2O2)-induced damage on human neuroblastoma cell line (SH-SY5Y) are associated with synaptic plasticity. The concentration screening of AS-IV and H2O2 on SH-SY5Y cells and the protective effects of AS-IV on SH-SY5Y cells under H2O2 stress were all determined by MTT assay. The expression of postsynaptic density 95 (PSD-95) and growth-associated protein 43 (GAP-43) were measured by western blot (WB) and inmunofluorescence staining assay under the same treatment conditions. According to the MTT results, the concentration of H2O2 at 50 μmol/L for 3 h was used for the cell damage model, and various concentrations of AS-IV (0.1, 0.2, 0.3, and 0.4 μmol/L) were used to affect SH-SY5Y cells. The MTT results showed that pretreatment of SH-SY5Y cells with AS-IV (0.1, 0.2, 0.3, and 0.4 μmol/L) attenuated the damage induced by H2O2 (50 μmol/L, 51.62% cell viability) and increased cell viability to 64.19, 63.48, 65.86, and 65.81%, respectively. Western blot analysis and immunofluorescence staining showed that the protective effects of AS-IV against SH-SY5Y cell damage caused by H2O2 resulted in reduced expression of PSD-95 and increased expression of GAP-43 in comparison with the H2O2 treatment group. The conclusion shows that AS-IV protected SH-SY5Y cells and enhanced their viability under H2O2 stress. AS-IV may facilitate presynaptic and postsynaptic plasticity to exert protective effects against oxidative damage of SH-SY5Y cells.

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