Journal: Redox Biology

Article Title: Glucose-6-phosphate dehydrogenase alleviates epileptic seizures by repressing reactive oxygen species production to promote signal transducer and activator of transcription 1-mediated N-methyl-d-aspartic acid receptors inhibition

doi: 10.1016/j.redox.2024.103236

Figure Lengend Snippet: Expression and functionality of pivotal enzymes in the glycolytic and pentose phosphate pathways. (a–d) Representative immunoblots (a, c) and quantification (b, d) of metabolism enzymes in the glycolysis and pentose phosphate pathways from the cortex and hippocampus of sham and chronic epileptic mice (n = 6). Data are presented as means ± SEM. **P < 0.01, ***P < 0.001, unpaired two-tailed Student's t-test. (e) Quantitative real-time polymerase chain reaction analysis of the expression of G6pd mRNA in the hippocampus at different time points after induction of status epilepticus (SE) (n = 5). Data are presented as means ± SEM. **P < 0.01, ***P < 0.001, one-way ANOVA with Tukey's post hoc test. (f) Relative enzyme activity of G6PD in the hippocampus at different time points after induction of SE (n = 3). Data are presented as means ± SEM. **P < 0.01, ***P < 0.001, one-way ANOVA with Tukey's post hoc test.

Article Snippet: For G6PD activator administration, the mice received bilateral intrahippocampal injections of AG1 (HY-123962; MCE, USA) at 4 μg or corn oil before intraperitoneal injection PTZ.

Techniques: Expressing, Western Blot, Two Tailed Test, Real-time Polymerase Chain Reaction, Activity Assay

Journal: Redox Biology

Article Title: Glucose-6-phosphate dehydrogenase alleviates epileptic seizures by repressing reactive oxygen species production to promote signal transducer and activator of transcription 1-mediated N-methyl-d-aspartic acid receptors inhibition

doi: 10.1016/j.redox.2024.103236

Figure Lengend Snippet: Localization of G6PD in epileptic brain tissues . (a–b) Immunostaining for G6PD with neuron-specific nucleoprotein (NeuN), glial fibrillary acidic protein (GFAP), and ionized calcium-binding adapter molecule 1 (Iba-1) in the sham and kainic acid (KA) groups at 28 d. Scale bars: 50 μm. (b–c) Quantitative analysis for the relative intensity of G6PD, NeuN, GFAP, and Iba-1 in single cells in the hippocampus CA3 region of epileptic mice at 28 d after KA injection (n = 50 cells from nine sections of each group, from three mice). Data are presented as means ± SEM. ***P < 0.001, unpaired two-tailed Student's t-test.

Article Snippet: For G6PD activator administration, the mice received bilateral intrahippocampal injections of AG1 (HY-123962; MCE, USA) at 4 μg or corn oil before intraperitoneal injection PTZ.

Techniques: Immunostaining, Binding Assay, Injection, Two Tailed Test

Journal: Redox Biology

Article Title: Glucose-6-phosphate dehydrogenase alleviates epileptic seizures by repressing reactive oxygen species production to promote signal transducer and activator of transcription 1-mediated N-methyl-d-aspartic acid receptors inhibition

doi: 10.1016/j.redox.2024.103236

Figure Lengend Snippet: G6PD modulates epileptic seizure activity. (a) Immunofluorescence labeling of enhanced green fluorescent proteins in the hippocampal CA1 region transfected with AAV-eGFP-adG6PD (n = 1). (b–c) Immunoblotting images (b) and quantification (c) of G6PD protein levels in the hippocampus of mice infected with the indicated AAVs (n = 6). Data are presented as the means ± SEM, ***P < 0.001, unpaired two-tailed Student's t-test. (d, e) Representative baseline and chronic epileptic electroencephalogram recordings from the cortex of chronic epileptic (adCtrl group) and G6PD overexpression after chronic epilepsy modeling (adG6PD group). (f, g) Quantitative analysis of the daily number of spontaneous recurrent seizures and duration of seizure-like events in the KA-induced epilepsy model in the adCtrl (n = 11) and adG6PD (n = 8) groups. Data are presented as means ± SEM. **P < 0.01, ***P < 0.001, unpaired two-tailed Student's t-test. (h–j) Representative traces and a summary of the amplitudes and frequencies of spontaneous excitatory postsynaptic currents in the hippocampal CA1 region in the adCtrl (n = 3, 6 cells) and adG6PD (n = 3, 8 cells) groups. (k–m) Representative traces and summary of the amplitude and frequency of spontaneous inhibitory postsynaptic currents in the hippocampal CA1 region of the adCtrl (n = 3, 8 cells) and adG6PD (n = 3, 8 cells) groups. Data are presented as means ± SEM.**P < 0.01, ***P < 0.001, unpaired two-tailed Student's t-test. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: For G6PD activator administration, the mice received bilateral intrahippocampal injections of AG1 (HY-123962; MCE, USA) at 4 μg or corn oil before intraperitoneal injection PTZ.

Techniques: Activity Assay, Immunofluorescence, Labeling, Transfection, Western Blot, Infection, Two Tailed Test, Over Expression

Journal: Redox Biology

Article Title: Glucose-6-phosphate dehydrogenase alleviates epileptic seizures by repressing reactive oxygen species production to promote signal transducer and activator of transcription 1-mediated N-methyl-d-aspartic acid receptors inhibition

doi: 10.1016/j.redox.2024.103236

Figure Lengend Snippet: G6PD negatively regulates the expression of N-methyl- d -aspartic acid receptors (NMDARs) through increasing STAT1 (a) RNA sequencing revealed that G6PD increased the transcription of Stat1 in epileptic mice. (b–e) Representative immunoblots (b, d) and quantification (c, e) of total STAT1 (n = 6), STAT3 (n = 6), p-STAT1 (n = 3), and nuclear STAT1 (n = 6) expression in the hippocampi of mice from the adCtrl and adG6PD groups. (f–i) Representative immunoblots (f, h) and quantification (g, i) of total and surface protein expression of NMDARs in the mouse hippocampus from the adCtrl and adG6PD groups (n = 6). Data are presented as means ± SEM. **P < 0.01, ***P < 0.001, unpaired two-tailed Student's t-test. (j) Effect of G6PD overexpression on the number of daily spontaneous recurrent seizures in the adCtrl, adG6PD, and adG6PD + fludarabine groups (n = 8). Data are presented as means ± SEM. ***P < 0.001, one-way ANOVA with Tukey's post-hoc test. (k–l) Representative immunoblots (b) and quantification (c) of NMDARs (Glu2A, Glu2B, and GluN1) in the hippocampi of mice in the adCtrl, adG6PD, and adG6PD + fludarabine groups (n = 6). Data are presented as means ± SEM. *P < 0.05, ***P < 0.001, one-way ANOVA with Tukey's post-hoc test.

Article Snippet: For G6PD activator administration, the mice received bilateral intrahippocampal injections of AG1 (HY-123962; MCE, USA) at 4 μg or corn oil before intraperitoneal injection PTZ.

Techniques: Expressing, RNA Sequencing, Western Blot, Two Tailed Test, Over Expression

Journal: Redox Biology

Article Title: Glucose-6-phosphate dehydrogenase alleviates epileptic seizures by repressing reactive oxygen species production to promote signal transducer and activator of transcription 1-mediated N-methyl-d-aspartic acid receptors inhibition

doi: 10.1016/j.redox.2024.103236

Figure Lengend Snippet: G6PD promotes the expression of STAT1 by decreasing reactive oxygen species (ROS) (a–d) Quantification of ROS, NADPH/NADP + , and GSH/GSSH in the control, AG1, AG1+DMF, 6-An, and 6-An + NAC groups after kainic acid (KA) treatment of HT22 cells. Data are presented as means ± SEM. ****P < 0.0001, one-way ANOVA with Tukey's post-hoc test. (e–h) Quantification of ROS, NADPH/NADP + , and GSH/GSSH in the control, 6-An, 6-An, 6-An, and 6-An + NAC groups after kainic acid (KA) treatment of HT22 cells. Data are presented as means ± SEM. ****P < 0.0001, one-way ANOVA with Tukey's post-hoc test. (i–i) Representative immunoblots (i) and quantification (j) of STAT1 in agonist (Con, AG1, and AG1+DMF groups) and inhibitor (Con, 6-An, and 6-An + NAC groups) groups after KA treatment of HT22 cells. Data are presented as means ± SEM. **P < 0.01, ***P < 0.001, one-way ANOVA with Tukey's post-hoc test.

Article Snippet: For G6PD activator administration, the mice received bilateral intrahippocampal injections of AG1 (HY-123962; MCE, USA) at 4 μg or corn oil before intraperitoneal injection PTZ.

Techniques: Expressing, Control, Western Blot

Journal: Redox Biology

Article Title: Glucose-6-phosphate dehydrogenase alleviates epileptic seizures by repressing reactive oxygen species production to promote signal transducer and activator of transcription 1-mediated N-methyl-d-aspartic acid receptors inhibition

doi: 10.1016/j.redox.2024.103236

Figure Lengend Snippet: Transcription factors Znf768 and Klf6 r egulate G6PD expression. (a) Quantitative real-time polymerase chain reaction analysis of predicted transcription factors (n = 4). Data are presented as means ± SEM, ns P > 0.05, **P < 0.01, nonsignificant unpaired two-tailed Student's t-test. (b–c) Representative immunoblots (b) and quantification (c) of ZNF768 and KLF6 from the hippocampus of sham and chronic epileptic mice (n = 6). Data are presented as means ± SEM, **P < 0.01, ***P < 0.001, unpaired two-tailed Student's t-test. (d) Relative luciferase activities in HEK293 cells. Cells were transiently co-transfected G6pd with vector or Znf768 or Klf6 (n = 6). Data are presented as means ± SEM, **P < 0.001, unpaired two-tailed Student's t-test.

Article Snippet: For G6PD activator administration, the mice received bilateral intrahippocampal injections of AG1 (HY-123962; MCE, USA) at 4 μg or corn oil before intraperitoneal injection PTZ.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Two Tailed Test, Western Blot, Luciferase, Transfection, Plasmid Preparation

Journal: Redox Biology

Article Title: Glucose-6-phosphate dehydrogenase alleviates epileptic seizures by repressing reactive oxygen species production to promote signal transducer and activator of transcription 1-mediated N-methyl-d-aspartic acid receptors inhibition

doi: 10.1016/j.redox.2024.103236

Figure Lengend Snippet: The G6PD activator AG1 alleviates seizure susceptibility (a) Example traces matching the minimal and tonic-clonic seizures of PTZ-injected mice. Scale bars, 0.3 mV and 1 s. (b) Timeline of 6-An and AG1 injection before the administration of pentylenetetrazol (PTZ) (12.5 mg/kg PTZ was injected into C57BL/6 mice once every 10 min, up to 10 times). (c) The seizure susceptibility in the sham and 6-An group following the injection of PTZ (n = 8). Data are presented as means ± SEM, **P < 0.01, unpaired two-tailed Student's t-test. (f, g) Immunoblot images and statistical analysis of hippocampal G6PD after AG1 administration (n = 4). Data are presented as means ± SEM. *P < 0.05, one-way ANOVA with Tukey's post-hoc test. (h, i) The seizure susceptibility in the sham and AG1 groups following the injection of PTZ (n = 8). Data are presented as means ± SEM, *P < 0.05, unpaired two-tailed Student's t-test.

Article Snippet: For G6PD activator administration, the mice received bilateral intrahippocampal injections of AG1 (HY-123962; MCE, USA) at 4 μg or corn oil before intraperitoneal injection PTZ.

Techniques: Injection, Two Tailed Test, Western Blot