Specific combinations of ion channel inhibitors reduce excessive Ca2+ influx as a consequence of oxidative stress and increase neuronal and glial cell viability in vitro

Objective: Ischemia-reperfusion (I/R) injury is a pathological feature of ischemic stroke. This study investigated the regulatory role of miR-485-5p in I/R injury. Methods: SH-SY5Y cells were induced with oxygen and glucose deprivation/reoxygenation (OGD/R) to mimic I/R injury in vitro. Cells were transfected with designated constructs (miR-485- 5p mimics, miR-485-5p inhibitor, lentiviral vectors overexpressing Rac1 or their corresponding controls). Cell viability was evaluated using the MTT assay. The concentrations of lactate dehydrogenase, malondialdehyde, and reactive oxygen species were detected to indicate the degree of oxidative stress. Flow cytometry and caspase-3 activity assay were used for apoptosis assessment. Dual-luciferase reporter assay was performed to confirm that Rac family small GTPase 1 (Rac1) was a downstream gene of miR-485-5p. Results: OGD/R resulted in decreased cell viability, elevated oxidative stress, increased apoptosis, and downregulated miR-485-5p expression in SH-SY5Y cells. MiR-485-5p upregulation alleviated I/R injury, evidenced by improved cell viability, decreased oxidative markers, and reduced apoptotic rate. OGD/R increased the levels of Rac1 and neurogenic locus notch homolog protein 2 (Notch2) signaling-related proteins in cells with normal miR-485-5p expression, whereas miR- 485-5p overexpression successfully suppressed OGD/R-induced upregulation of these proteins. Furthermore, the delivery of vectors overexpressing Rac1 in miR-485-5p mimics-transfected cells reversed the protective effect of miR-485-5p in cells with OGD/R-induced injury. Conclusion: This study showed that miR-485-5p protected cells following I/R injury via targeting Rac1/Notch2 signaling suggest that targeted upregulation of miR-485-5p might be a promising therapeutic option for the protection against I/R injury.

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Inhibitory effects of Panax ginseng glycoproteins in models of doxorubicin-induced cardiac toxicity in vivo and in vitro

Food & Function ◽

10.1039/d1fo01307f ◽

2021 ◽

Author(s):

Yajun Chen ◽

Lei Wang ◽

Tianjia Liu ◽

Zhidong Qiu ◽

Ye Qiu ◽

...

Keyword(s):

Oxidative Stress ◽

Protective Effect ◽

Cell Viability ◽

Panax Ginseng ◽

Cardiac Toxicity ◽

H9c2 Cell ◽

Inhibitory Effects ◽

Myocardial Oxidative Stress

We investigated the protective effect of PGP against DOX-induced cardiotoxicity in vitro and in vivo. PGP increases H9C2 cell viability and inhibits apoptosis, alleviating DOX-induced myocardial oxidative stress-related cardiotoxicity.

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In vitro effect of vanadyl sulfate on cultured primary astrocytes: cell viability and oxidative stress markers.

Journal of Applied Toxicology ◽

10.1002/jat.3939 ◽

2020 ◽

Vol 40 (6) ◽

pp. 737-747

Author(s):

Agnieszka Ścibior ◽

Konrad A. Szychowski ◽

Iwona Zwolak ◽

Klaudia Dachowska ◽

Jan Gmiński

Keyword(s):

Oxidative Stress ◽

Cell Viability ◽

Vanadyl Sulfate ◽

Oxidative Stress Markers ◽

Stress Markers ◽

Primary Astrocytes ◽

Vitro Effect ◽

And Oxidative Stress

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TrxR2 overexpression alleviates inflammation-mediated neuronal death via reducing the oxidative stress and activating the Akt–Parkin pathway

Toxicology Research ◽

10.1039/c9tx00076c ◽

2019 ◽

Vol 8 (5) ◽

pp. 641-653 ◽

Cited By ~ 1

Author(s):

Jinbao Gao ◽

Yunjun Li ◽

Wende Li ◽

Haijiang Wang

Keyword(s):

Oxidative Stress ◽

Cell Death ◽

Cell Viability ◽

Western Blotting ◽

Neuronal Death ◽

Protective Effects ◽

Mitochondrial Apoptosis ◽

Inflammation Response ◽

N2a Cells

Abstract Neuronal death caused by inflammatory cytokine-mediated neuroinflammation is being extensively explored. Thioredoxin reductase (TrxR) 2 is a novel mediator of inflammation response. In the current study, we focus on the mechanisms of TrxR2 overexpression in inflammation-mediated neuronal death. LPS was used to induce neuroinflammation in N2a cells in vitro. Adenovirus-loaded TrxR2 was transfected into N2a cells to up-regulate TrxR2 expression. Then, cell viability was determined via MTT assay and TUNEL assay. Apoptosis was measured via western blotting and ELISA. Oxidative stress was detected via ELISA and flow cytometry. A pathway inhibitor was used to verify the role of the Akt–Parkin pathway in the LPS-mediated N2a cell death in the presence of TrxR2 overexpression. With the help of immunofluorescence assay and western blotting, we found that TrxR2 expression was significantly reduced in response to LPS treatment, and this effect was associated with N2a cell death via apoptosis. At the molecular level, TrxR2 overexpression elevated the activity of the Akt–Parkin pathway, as evidenced by the increased expression of p-Akt and Parkin. Interestingly, inhibition of the Akt–Parkin pathway abolished the regulatory effect of TrxR2 on LPS-treated N2a cells, as evidenced by the decreased cell viability and increased apoptotic ratio. Besides, TrxR2 overexpression also reduced oxidative stress, inflammation factor transcription and mitochondrial apoptosis. However, inhibition of Akt–Parkin axis abrogated the protective effects of TrxR2 on redox balance, mitochondrial performance and cell survival. LPS-mediated neuronal death was linked to a drop in TrxR2 overexpression and the inactivation of the Akt–Parkin pathway. Overexpression of TrxR2 sustained mitochondrial function, inhibited oxidative stress, repressed inflammation response, and blocked mitochondrial apoptosis, finally sending a pro-survival signal for the N2a cells in the setting of LPS-mediated inflammation environment.

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Cytochrome c oxidase III as a mechanism for apoptosis in heart failure following myocardial infarction

AJP Cell Physiology ◽

10.1152/ajpcell.00045.2009 ◽

2009 ◽

Vol 297 (4) ◽

pp. C928-C934 ◽

Cited By ~ 23

Author(s):

Changgong Wu ◽

Lin Yan ◽

Christophe Depre ◽

Sunil K. Dhar ◽

You-Tang Shen ◽

...

Keyword(s):

Oxidative Stress ◽

Myocardial Infarction ◽

Heart Failure ◽

Cytochrome C ◽

Protein Expression ◽

Cell Viability ◽

Cytochrome C Oxidase ◽

Mitochondrial Gene

Cytochrome c oxidase (COX) is composed of 13 subunits, of which COX I, II, and III are encoded by a mitochondrial gene. COX I and II function as the main catalytic components, but the function of COX III is unclear. Because myocardial ischemia affects mitochondrial oxidative metabolism, we hypothesized that COX activity and expression would be affected during postischemic cardiomyopathy. This hypothesis was tested in a monkey model following myocardial infarction (MI) and subsequent pacing-induced heart failure (HF). In this model, COX I protein expression was decreased threefold after MI and fourfold after HF ( P < 0.05 vs. sham), whereas COX II expression remained unchanged. COX III protein expression increased 5-fold after MI and further increased 10-fold after HF compared with sham ( P < 0.05 vs. sham). The physiological impact of COX III regulation was examined in vitro. Overexpression of COX III in mitochondria of HL-1 cells resulted in an 80% decrease in COX I, 60% decrease in global COX activity, 60% decrease in cell viability, and threefold increase in apoptosis ( P < 0.05). Oxidative stress induced by H2O2 significantly ( P < 0.05) increased COX III expression. H2O2 decreased cell viability by 47 ± 3% upon overexpression of COX III, but only by 12 ± 5% in control conditions ( P < 0.05). We conclude that ischemic stress in vivo and oxidative stress in vitro lead to upregulation of COX III, followed by downregulation of COX I expression, impaired COX oxidative activity, and increased apoptosis. Therefore, upregulation of COX III may contribute to the increased susceptibility to apoptosis following MI and subsequent HF.

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Differential concentration-specific effects of caffeine on cell viability, oxidative stress, and cell cycle in pulmonary oxygen toxicity in vitro

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2014.06.132 ◽

2014 ◽

Vol 450 (4) ◽

pp. 1345-1350 ◽

Cited By ~ 31

Author(s):

Kirti Kumar Tiwari ◽

Chun Chu ◽

Xanthi Couroucli ◽

Bhagavatula Moorthy ◽

Krithika Lingappan

Keyword(s):

Oxidative Stress ◽

Cell Cycle ◽

Cell Viability ◽

Oxygen Toxicity ◽

Specific Effects ◽

Toxicity In Vitro

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Protective Effect of Liposome-Encapsulated Glutathione in a Human Epidermal Model Exposed to a Mustard Gas Analog

Journal of Toxicology ◽

10.1155/2011/109516 ◽

2011 ◽

Vol 2011 ◽

pp. 1-11 ◽

Cited By ~ 17

Author(s):

Victor Paromov ◽

Sudha Kumari ◽

Marianne Brannon ◽

Naga S. Kanaparthy ◽

Hongsong Yang ◽

...

Keyword(s):

Oxidative Stress ◽

Protective Effect ◽

Cell Viability ◽

Sulfur Mustard ◽

Alkylating Agents ◽

Water Soluble ◽

Model Systems ◽

Mustard Gas

Sulfur mustard or mustard gas (HD) and its monofunctional analog, 2-chloroethyl ethyl sulfide (CEES), or “half-mustard gas,” are alkylating agents that induce DNA damage, oxidative stress, and inflammation. HD/CEES are rapidly absorbed in the skin causing extensive injury. We hypothesize that antioxidant liposomes that deliver both water-soluble and lipid-soluble antioxidants protect skin cells from immediate CEES-induced damage via attenuating oxidative stress. Liposomes containing water-soluble antioxidants and/or lipid-soluble antioxidants were evaluated usingin vitromodel systems. Initially, we found that liposomes containing encapsulated glutathione (GSH-liposomes) increased cell viability and attenuated production of reactive oxygen species (ROS) in HaCaT cells exposed to CEES. Next, GSH-liposomes were tested in a human epidermal model, EpiDerm. In the EpiDerm, GSH-liposomes administered simultaneously or 1 hour after CEES exposure (2.5 mM) increased cell viability, inhibited CEES-induced loss of ATP and attenuated changes in cellular morphology, but did not reduce caspase-3 activity. These findings paralleled the previously describedin vivoprotective effect of antioxidant liposomes in the rat lung and established the effectiveness of GSH-liposomes in a human epidermal model. This study provides a rationale for use of antioxidant liposomes against HD toxicity in the skin considering further verification in animal models exposed to HD.

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Oxytocin Promotes C6 Glial Cell Death and Aggravates Hydrogen Peroxide-induced Oxidative Stress

10.36922/itps.v3i1.939 ◽

2020 ◽

pp. 27-33

Author(s):

Ahmet Sevki Taskiran ◽

Merve Ergul

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

Cell Death ◽

Cell Viability ◽

Glial Cell ◽

Glial Cells ◽

Control Group ◽

C6 Cells ◽

Cox 2 ◽

Cox 1

Background. Recent studies have shown that oxytocin plays a vital role in neurons and glial cells. However, its effect on hydrogen peroxide (H2O2)-induced oxidative stress as well as cyclooxygenase-1 (COX-1) and COX-2 in glial cells are still unclear. Objective. This study aims to examine the effect of oxytocin on glial cell viability, oxidative stress, COX-1, and COX-2 in C6 glial cells after exposure to H2O2. Methods. In this study, C6 glioma cell line was used to study the effect of oxytocin on the glial cell in four cell groups. The control group was untreated. Cells in the H2O2 group were treated with 0.5 mM H2O2 for 24 h. Cells in the oxytocin group were treated with various concentrations (0.25, 0.5, 1, and 2 μg/mL) of oxytocin for 24 h. Cells in the oxytocin+H2O2 group were pre-treated with various concentrations (0.25, 0.5, 1, and 2 μg/mL) of oxytocin for 1 h before 24-h exposure to 0.5 mM H2O2. Cell viability was evaluated using XTT assay. Total antioxidant status and total oxidant status (TOS), COX-1, and COX-2 levels in the cells were measured by commercial kits. Results. Oxytocin with various concentrations significantly decreased the viability of C6 cells after H2O2-induced oxidative stress (P < 0.01). It also significantly increased the levels of TOS, COX-1, and COX-2 in C6 cells after H2O2-induced oxidative stress (P < 0.001). Conclusion. Oxytocin increases glial cell death after H2O2-induced oxidative damage in C6 cells, along with upregulation of COX-1 and COX-2 levels.

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Targeted Delivery of Intranasally Administered Cyclovirobuxine D Liposomes to Brain for Treatment Cerebral Ischemia-Reperfusion Injury via Anti-oxidative Stress and Activating Autophagy

10.21203/rs.3.rs-966870/v2 ◽

2022 ◽

Author(s):

Tuo Liu ◽

Fang Yang ◽

Xiangyi Lu ◽

Chang Liu ◽

Yang Yu ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Viability ◽

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Cerebral Ischemia Reperfusion ◽

Protein Expressions ◽

Cerebral Ischemia Reperfusion Injury ◽

Ht22 Cell

Abstract The lack of effective therapy mandates development of treatment for cerebral ischemia-reperfusion injury (CIRI. The previous study suggested that Cyclovirobuxine D (CVBD) encapsulated in Angiopep-conjugated Polysorbate 80-Coated Liposomes showed a better brain targeting by intranasal administration. Therefore, this study focused on the protection and mechanism of CVBD brain-targeted liposomes in treating CIRI. In order to evaluate these, the CIRI rat model was induced by middle cerebral artery occlusion (MCAO)-reperfusion. Pharmacological evaluation was assessed in vivo by general indexs, neurobehavioral scores, triphenyl tetrazolium chloride (TTC) staining, histopathological staining (HE staining and Nissl staining), small animal magnetic resonance imaging, biochemical assay and Western blot. The results show that CVBD liposomes alleviated pathological damage of brain. Futhermore, the protective effect of CVBD liposomes on OGD/R-injured HT22 cell was investigated by cell fusion degree, cell proliferation curve and cell viability. OGD/R-injured HT22 cell was infected by mRFP-GFP-LC3 adenovirus. The autophagosome and autophagy flow were observed by laser confocal microscopy, and autophagy-related protein expressions (LC3, p62 and Beclin 1) were analyzed by Western blot. Meanwhile, the classic autophagy inhibitor, chloroquine, was used to explore the autophagy-regulated mechanism of CVBD brain-targeted liposomes in treating CIRI. In cell model of oxygen and glucose deprivation/re-oxygenation, CVBD liposomes increased cell viability and decreased ROS level. CVBD liposomes improved oxidative stress protein expressions and activated autophagy in vitro. Furthermore, CVBD liposomes reversed the decrease of cell viability, increase of ROS level, and reduction of protein expressions associated to anti-oxidative stress and autophagy induced by chloroquine. Collectively, CVBD liposomes inhibited CIRI via regulating oxidative stress and enhancing autophagy level in vivo and in vitro, showing a great potential in treating CIRI in clinic.

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EVALUATION OF TAURINE’S NEUROPROTECTIVE EFFECTS ON SH-SY5Y CELLS UNDER OXIDATIVE STRESS

10.1101/2021.04.26.441504 ◽

2021 ◽

Author(s):

Rafaella Carvalho Rossato ◽

Alessandro Eustaquio Campos Granato ◽

Jessica Cristina Pinto ◽

Carlos Dailton Guedes de Oliveira Moraes ◽

Geisa Nogueira Salles ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Viability ◽

Cell Morphology ◽

Neuroprotective Effect ◽

Study Strategies ◽

Neuroprotective Effects ◽

Human Neuroblastoma ◽

Characteristic Model ◽

The Face

ABSTRACTAlzheimer’s disease (AD) is a type of dementia that affects millions of people. Although there is no cure, several study strategies seek to elucidate the mechanisms of the disease. Recent studies address the benefits of taurine. Thus, the present study aims to analyze the neuroprotective effect of taurine on human neuroblastoma, using an in vitro experimental model of oxidative stress induced by hydrocortisone in the SH-SY5Y cell line as a characteristic model of AD. The violet crystal assay was used for cell viability and the evaluation of cell morphology was performed by scanning electron microscopy (SEM). After pretreatment with taurine, the SH-SY5Y cell showed an improvement in cell viability in the face of oxidative stress and improved cell morphology. Thus, the treatment presented a neuroprotective effect.GRAPHICAL ABSTRACT

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