medchemexpress llc  (MedChemExpress)

Journal: Neural Regeneration Research

Article Title: Hypoxia-inducible factor 1α inhibitor reduces hippocampal neuronal ferroptosis

doi: 10.4103/NRR.NRR-D-24-01007

Figure Lengend Snippet: Experimental design. Each behavioral testing and molecular biology experiment was repeated at least three times. CON: Control; DHE: dihydroethidium; ELISA: enzyme linked immunosorbent assay; HE: hematoxylin-eosin; NS: normal saline; PTZ: pentylenetetrazol; PX-478: a hypoxia-inducible factor-1α inhibitor; qPCR: quantitative reverse transcription polymerase chain reaction; TEM: transmission electron microscopy; WB: western blot.

Article Snippet: Using a random number table, the mice were randomly assigned into four groups ( n = 16 per group) ( ): (1) CON group: received two intraperitoneal injections of normal saline (0.2 mL), 30 minutes apart, every other day, for a total of 11 times; (2) PTZ group: received an intraperitoneal injection of normal saline (0.2 mL) followed by 35 mg/kg PTZ (Sigma-Aldrich, St. Louis, MO, USA, P6500) 30 minutes later, every other day, for a total of 11 times; (3) PTZ + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 (MedChemExpress, Monmouth Junction, NJ, USA, HY-10231) followed by 35 mg/kg PTZ 30 minutes later, every other day, for a total of 11 times; (4) CON + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 followed by normal saline (0.2 mL) 30 minutes later, every other day, for a total of 11 times.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Saline, Reverse Transcription, Polymerase Chain Reaction, Transmission Assay, Electron Microscopy, Western Blot

Journal: Neural Regeneration Research

Article Title: Hypoxia-inducible factor 1α inhibitor reduces hippocampal neuronal ferroptosis

doi: 10.4103/NRR.NRR-D-24-01007

Figure Lengend Snippet: Seizure scores after each injection of PTZ in mice. Adult male C57BL/6 mice were assigned to four experimental groups: the CON group received saline only; the PTZ group received saline followed by 35 mg/kg of PTZ; the PTZ + PX-478 group received 5 mg/kg of PX-478 before PTZ administration; and the CON + PX-478 group received PX-478 followed by normal saline. Data are expressed as mean ± SEM ( n = 16). * P < 0.05, *** P < 0.001, PTZ vs . PTZ + PX-478 (Student’s t -test). CON: Control; PTZ: pentylenetetrazol; PX-478: a hypoxia-inducible factor-1α inhibitor.

Article Snippet: Using a random number table, the mice were randomly assigned into four groups ( n = 16 per group) ( ): (1) CON group: received two intraperitoneal injections of normal saline (0.2 mL), 30 minutes apart, every other day, for a total of 11 times; (2) PTZ group: received an intraperitoneal injection of normal saline (0.2 mL) followed by 35 mg/kg PTZ (Sigma-Aldrich, St. Louis, MO, USA, P6500) 30 minutes later, every other day, for a total of 11 times; (3) PTZ + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 (MedChemExpress, Monmouth Junction, NJ, USA, HY-10231) followed by 35 mg/kg PTZ 30 minutes later, every other day, for a total of 11 times; (4) CON + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 followed by normal saline (0.2 mL) 30 minutes later, every other day, for a total of 11 times.

Techniques: Injection, Saline, Control

Journal: Neural Regeneration Research

Article Title: Hypoxia-inducible factor 1α inhibitor reduces hippocampal neuronal ferroptosis

doi: 10.4103/NRR.NRR-D-24-01007

Figure Lengend Snippet: Effect of PX-478 on neuronal damage in hippocampal CA1, CA3, and DG regions of epilepsy model mice. In the CON and CON + PX-478 groups, neurons in the hippocampal CA1, CA3, and DG regions were organized and intact. In the PTZ group, neurons in CA1 and CA3 showed disorganized arrangement, loss, and abnormal morphology. DG damage was less severe. The PTZ + PX-478 group exhibited reduced neuronal damage in CA1 and CA3 compared with that in the PTZ group, though some neuronal loss remained. Scale bars: 100 μm. CON: Control; DG: dentate gyrus; PTZ: pentylenetetrazol; PX-478: a hypoxia-inducible factor-1α inhibitor.

Article Snippet: Using a random number table, the mice were randomly assigned into four groups ( n = 16 per group) ( ): (1) CON group: received two intraperitoneal injections of normal saline (0.2 mL), 30 minutes apart, every other day, for a total of 11 times; (2) PTZ group: received an intraperitoneal injection of normal saline (0.2 mL) followed by 35 mg/kg PTZ (Sigma-Aldrich, St. Louis, MO, USA, P6500) 30 minutes later, every other day, for a total of 11 times; (3) PTZ + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 (MedChemExpress, Monmouth Junction, NJ, USA, HY-10231) followed by 35 mg/kg PTZ 30 minutes later, every other day, for a total of 11 times; (4) CON + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 followed by normal saline (0.2 mL) 30 minutes later, every other day, for a total of 11 times.

Techniques: Control

Journal: Neural Regeneration Research

Article Title: Hypoxia-inducible factor 1α inhibitor reduces hippocampal neuronal ferroptosis

doi: 10.4103/NRR.NRR-D-24-01007

Figure Lengend Snippet: Effect of PX-478 on the expression of oxidative stress indicators in the hippocampus of epilepsy model mice. (A) ROS staining of CA1, CA3, and DG regions in hippocampus. Compared with the CON and CON + PX-478 groups, the PTZ group appeared to have higher ROS levels in all regions, which were reduced in the PTZ + PX-478 group. Scale bars: 100 μm. (B) Relative activity of antioxidant enzymes SOD, CAT, and GSH-Px in hippocampus. (C) Relative MDA content in hippocampus. The data, normalized to CON, are presented as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s honestly significant difference test). CAT: Catalase; CON: control; DG: dentate gyrus; GSH-Px: glutathione peroxidase; MDA: malondialdehyde; ns: not significant; PTZ: pentylenetetrazol; PX-478: a hypoxia-inducible factor-1α inhibitor; ROS: reactive oxygen species; SOD: superoxide dismutase.

Article Snippet: Using a random number table, the mice were randomly assigned into four groups ( n = 16 per group) ( ): (1) CON group: received two intraperitoneal injections of normal saline (0.2 mL), 30 minutes apart, every other day, for a total of 11 times; (2) PTZ group: received an intraperitoneal injection of normal saline (0.2 mL) followed by 35 mg/kg PTZ (Sigma-Aldrich, St. Louis, MO, USA, P6500) 30 minutes later, every other day, for a total of 11 times; (3) PTZ + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 (MedChemExpress, Monmouth Junction, NJ, USA, HY-10231) followed by 35 mg/kg PTZ 30 minutes later, every other day, for a total of 11 times; (4) CON + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 followed by normal saline (0.2 mL) 30 minutes later, every other day, for a total of 11 times.

Techniques: Expressing, Staining, Activity Assay, Control

Journal: Neural Regeneration Research

Article Title: Hypoxia-inducible factor 1α inhibitor reduces hippocampal neuronal ferroptosis

doi: 10.4103/NRR.NRR-D-24-01007

Figure Lengend Snippet: Effect of PX-478 on mitochondrial damage in hippocampal neurons of epilepsy model mice. (A) Transmission electron microscope images of hippocampus for each group with 5.0 μm, 2.0 μm, and 500 nm scales. In the CON and CON + PX-478 groups, the morphology and size of mitochondria in hippocampal neurons appeared normal. In the PTZ group, hippocampal neurons displayed smaller mitochondria with increased membrane density and reduced or absent cristae, whereas the PTZ + PX-478 group showed a notable reduction in mitochondrial damage. Arrows indicate mitochondria. (B) Quantification of mitochondrial length and area in the hippocampus. The data are presented as mean ± SEM ( n = 5). * P < 0.05, *** P < 0.001 (one-way analysis of variance followed by Tukey’s honest significant difference test). CON: Control; ns: not significant; PTZ: pentylenetetrazol; PX-478: a hypoxia-inducible factor-1α inhibitor.

Article Snippet: Using a random number table, the mice were randomly assigned into four groups ( n = 16 per group) ( ): (1) CON group: received two intraperitoneal injections of normal saline (0.2 mL), 30 minutes apart, every other day, for a total of 11 times; (2) PTZ group: received an intraperitoneal injection of normal saline (0.2 mL) followed by 35 mg/kg PTZ (Sigma-Aldrich, St. Louis, MO, USA, P6500) 30 minutes later, every other day, for a total of 11 times; (3) PTZ + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 (MedChemExpress, Monmouth Junction, NJ, USA, HY-10231) followed by 35 mg/kg PTZ 30 minutes later, every other day, for a total of 11 times; (4) CON + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 followed by normal saline (0.2 mL) 30 minutes later, every other day, for a total of 11 times.

Techniques: Transmission Assay, Microscopy, Membrane, Control

Journal: Neural Regeneration Research

Article Title: Hypoxia-inducible factor 1α inhibitor reduces hippocampal neuronal ferroptosis

doi: 10.4103/NRR.NRR-D-24-01007

Figure Lengend Snippet: Effect of PX-478 on the expression of ferroptosis indicators in the hippocampus of epilepsy model mice. (A) Relative Fe 2+ accumulation in the hippocampus. (B–D) RT-qPCR and western blot results for GPX4 (B), PTGS2/COX-2 (C), and FTH-1 and TRF (D) in the hippocampus. The data are presented as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s honest significant difference test). CON: Control; COX-2: cyclooxygenase 2; FTH-1: ferritin heavy chain 1; GPX4: glutathione peroxidase 4; ns: not significant; PTGS2: prostaglandin synthase 2; PTZ: pentylenetetrazol; PX-478: a hypoxia-inducible factor-1α inhibitor; RT-qPCR: reverse transcription quantitative polymerase chain reaction; TRF: transferrin.

Article Snippet: Using a random number table, the mice were randomly assigned into four groups ( n = 16 per group) ( ): (1) CON group: received two intraperitoneal injections of normal saline (0.2 mL), 30 minutes apart, every other day, for a total of 11 times; (2) PTZ group: received an intraperitoneal injection of normal saline (0.2 mL) followed by 35 mg/kg PTZ (Sigma-Aldrich, St. Louis, MO, USA, P6500) 30 minutes later, every other day, for a total of 11 times; (3) PTZ + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 (MedChemExpress, Monmouth Junction, NJ, USA, HY-10231) followed by 35 mg/kg PTZ 30 minutes later, every other day, for a total of 11 times; (4) CON + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 followed by normal saline (0.2 mL) 30 minutes later, every other day, for a total of 11 times.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: Hypoxia-inducible factor 1α inhibitor reduces hippocampal neuronal ferroptosis

doi: 10.4103/NRR.NRR-D-24-01007

Figure Lengend Snippet: Effect of PX-478 on the expression of HIF-1α/HO-1 pathway in the hippocampus of epilepsy model mice. (A) RT-qPCR results for HIF-1α and HO-1 in the hippocampus. (B) Western blot results for HIF-1α and HO-1 in the hippocampus. The data were normalized to CON and presented as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s honest significant difference test). CON: Control; HIF-1α: hypoxia-inducible factor 1α; HO-1: heme oxygenase-1; ns: not significant; PTZ: pentylenetetrazol; PX-478: a hypoxia-inducible factor-1α inhibitor; RT-qPCR: reverse transcription quantitative polymerase chain reaction.

Article Snippet: Using a random number table, the mice were randomly assigned into four groups ( n = 16 per group) ( ): (1) CON group: received two intraperitoneal injections of normal saline (0.2 mL), 30 minutes apart, every other day, for a total of 11 times; (2) PTZ group: received an intraperitoneal injection of normal saline (0.2 mL) followed by 35 mg/kg PTZ (Sigma-Aldrich, St. Louis, MO, USA, P6500) 30 minutes later, every other day, for a total of 11 times; (3) PTZ + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 (MedChemExpress, Monmouth Junction, NJ, USA, HY-10231) followed by 35 mg/kg PTZ 30 minutes later, every other day, for a total of 11 times; (4) CON + PX-478 group: received an intraperitoneal injection of 5 mg/kg PX-478 followed by normal saline (0.2 mL) 30 minutes later, every other day, for a total of 11 times.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: LDHA protects vascular endothelial cells from oxidative stress-induced mitochondrial damage via HIF-1α activation and glycolytic reprogramming

doi: 10.3892/mmr.2026.13851

Figure Lengend Snippet: LDHA overexpression enhances HIF-1α expression and nuclear translocation in VECs under OGD/R conditions. (A) WB of total HIF-1α expression in VECs treated with 0.5 mM H 2 O 2 (OGD/R) and overexpressing LDHA. (B) Semi-quantification of total HIF-1α protein levels from WB normalized to β-actin. (C) Representative western blot analysis of cytoplasmic HIF-1α from fractionated lysates; β-actin was used as the cytoplasmic control. (D) Semi-quantification of cytoplasmic HIF-1α protein in (C) using β-actin as a standard. (E) Representative western blot analysis of nuclear HIF-1α; Lamin B1 was used as a nuclear control. (F) Semi-quantification of nuclear HIF-1α protein in (E) using Lamin B1 as a standard. (G) Immunofluorescence staining of HIF-1α localization in VECs treated with 0.5 mM H 2 O 2 (OGD/R) and overexpressing LDHA. Magnification, ×40. Data are presented as the mean ± SD of three independent experiments. *P<0.05. H 2 O 2 , hydrogen peroxide; OGD/R, oxygen-glucose deprivation/reperfusion; VECs, vascular endothelial cells; LDHA, lactate dehydrogenase A; HIF-1α, hypoxia-inducible factor 1α; WB, western blotting.

Article Snippet: To investigate the roles of HIF-1α signaling and glycolysis, the cells were treated with 20 μM PX478 (MedChemExpress), a HIF-1α inhibitor, for 20 h at 37°C, or 2 mM 2-deoxy-D-glucose (2-DG; MedChemExpress), a glycolysis inhibitor, for 24 h at 37°C, under oxidative stress conditions induced by preconditioning with 0.5 mM H 2 O 2 for 12 h at 37°C, followed by OGD/R (6-h OGD at 37°C and 24-h reoxygenation at 37°C) [hereafter referred to as 0.5 mM H 2 O 2 (OGD/R)].

Techniques: Over Expression, Expressing, Translocation Assay, Western Blot, Control, Immunofluorescence, Staining

Journal: Molecular Medicine Reports

Article Title: LDHA protects vascular endothelial cells from oxidative stress-induced mitochondrial damage via HIF-1α activation and glycolytic reprogramming

doi: 10.3892/mmr.2026.13851

Figure Lengend Snippet: LDHA overexpression alleviates OGD/R-induced apoptosis and mitochondrial damage in VECs via HIF-1α nuclear translocation. (A) Flow cytometric analysis of apoptosis in VECs treated with 0.5 mM H 2 O 2 (OGD/R) and overexpressing LDHA, with or without the HIF-1α inhibitor PX478. (B) ROS levels measured using a ROS assay kit in VECs treated with 0.5 mM H 2 O 2 (OGD/R) and overexpressing LDHA, with or without PX478. (C) MMP was assessed by JC-1 fluorescent dye in VECs treated with 0.5 mM H 2 O 2 (OGD/R) and overexpressing LDHA, with or without PX478. The images show representative red (intact MMP) and green (depolarized MMP) fluorescence. Magnification, ×40. (D-G) WB of cytoplasmic and nuclear HIF-1α expression in VECs treated with 0.5 mM H 2 O 2 (OGD/R) and overexpressing LDHA, with or without PX478. (D) Representative cytoplasmic HIF-1α western blot analysis; β-actin was used as a cytoplasmic control. (E) Semi-quantification of cytoplasmic HIF-1α in (D), normalized to β-actin. (F) Representative nuclear HIF-1α western blot analysis; Lamin B1 was used as a nuclear control. (G) Semi-quantification of nuclear HIF-1α in (F), normalized to Lamin B1. Data are presented as the mean ± SD of three independent experiments. *P<0.05 and **P<0.01 . H 2 O 2 , hydrogen peroxide; OGD/R, oxygen-glucose deprivation/reperfusion; VECs, vascular endothelial cells; WB, western blotting; PI, propidium iodide; FITC, fluorescein isothiocyanate; LDHA, lactate dehydrogenase A; HIF-1α, hypoxia-inducible factor 1α; ROS, reactive oxygen species; MMP, mitochondrial membrane potential.

Article Snippet: To investigate the roles of HIF-1α signaling and glycolysis, the cells were treated with 20 μM PX478 (MedChemExpress), a HIF-1α inhibitor, for 20 h at 37°C, or 2 mM 2-deoxy-D-glucose (2-DG; MedChemExpress), a glycolysis inhibitor, for 24 h at 37°C, under oxidative stress conditions induced by preconditioning with 0.5 mM H 2 O 2 for 12 h at 37°C, followed by OGD/R (6-h OGD at 37°C and 24-h reoxygenation at 37°C) [hereafter referred to as 0.5 mM H 2 O 2 (OGD/R)].

Techniques: Over Expression, Translocation Assay, ROS Assay, Fluorescence, Expressing, Western Blot, Control, Membrane

Journal: Molecular Medicine Reports

Article Title: LDHA protects vascular endothelial cells from oxidative stress-induced mitochondrial damage via HIF-1α activation and glycolytic reprogramming

doi: 10.3892/mmr.2026.13851

Figure Lengend Snippet: LDHA overexpression alleviates OGD/R-induced apoptosis and mitochondrial damage in VECs via HIF-1α nuclear translocation. (A) Flow cytometric analysis of apoptosis in VECs treated with 0.5 mM H 2 O 2 (OGD/R) and overexpressing LDHA, with or without the HIF-1α inhibitor PX478. (B) ROS levels measured using a ROS assay kit in VECs treated with 0.5 mM H 2 O 2 (OGD/R) and overexpressing LDHA, with or without PX478. (C) MMP was assessed by JC-1 fluorescent dye in VECs treated with 0.5 mM H 2 O 2 (OGD/R) and overexpressing LDHA, with or without PX478. The images show representative red (intact MMP) and green (depolarized MMP) fluorescence. Magnification, ×40. (D-G) WB of cytoplasmic and nuclear HIF-1α expression in VECs treated with 0.5 mM H 2 O 2 (OGD/R) and overexpressing LDHA, with or without PX478. (D) Representative cytoplasmic HIF-1α western blot analysis; β-actin was used as a cytoplasmic control. (E) Semi-quantification of cytoplasmic HIF-1α in (D), normalized to β-actin. (F) Representative nuclear HIF-1α western blot analysis; Lamin B1 was used as a nuclear control. (G) Semi-quantification of nuclear HIF-1α in (F), normalized to Lamin B1. Data are presented as the mean ± SD of three independent experiments. *P<0.05 and **P<0.01 . H 2 O 2 , hydrogen peroxide; OGD/R, oxygen-glucose deprivation/reperfusion; VECs, vascular endothelial cells; WB, western blotting; PI, propidium iodide; FITC, fluorescein isothiocyanate; LDHA, lactate dehydrogenase A; HIF-1α, hypoxia-inducible factor 1α; ROS, reactive oxygen species; MMP, mitochondrial membrane potential.

Article Snippet: To investigate the roles of HIF-1α signaling and glycolysis, the cells were treated with 20 μM PX478 (MedChemExpress), a HIF-1α inhibitor, for 20 h at 37°C, or 2 mM 2-deoxy-D-glucose (2-DG; MedChemExpress), a glycolysis inhibitor, for 24 h at 37°C, under oxidative stress conditions induced by preconditioning with 0.5 mM H 2 O 2 for 12 h at 37°C, followed by OGD/R (6-h OGD at 37°C and 24-h reoxygenation at 37°C) [hereafter referred to as 0.5 mM H 2 O 2 (OGD/R)].

Techniques: Over Expression, Translocation Assay, ROS Assay, Fluorescence, Expressing, Western Blot, Control, Membrane