Journal: Neurotoxicity research

Article Title: The MT1G Gene in LUHMES Neurons Is a Sensitive Biomarker of Neurotoxicity

doi: 10.1007/s12640-020-00272-3

Figure Lengend Snippet: Chemicals used and abbreviations

Article Snippet: Chemicals Chemicals included Ferbam from TCI America, Portland, OR; ML385 and MS275 from Toris Biosciences, Minneapolis, MN; APTO235 (MedChemExpress, Princeton, NJ); and sankel (Muse Chem, Fairfield, NJ).

Techniques:

Journal: Neurotoxicity research

Article Title: The MT1G Gene in LUHMES Neurons Is a Sensitive Biomarker of Neurotoxicity

doi: 10.1007/s12640-020-00272-3

Figure Lengend Snippet: MT1G mRNA expression responses to chelated and unchelated metals. a dLUHMES cells were treated for 6 h with toxicants MHG, CuCl2, DDC, or ziram; and with vehicle, white bars; APTO-253 5 μM, speckled bars; or ML385 5 μM, dark hatched bars. MT1G expression was measured by qRT-PCR. Treatments were MHG 2 μM; CuCl2 5 μM; DDC 5 μM; NiDDC2 5 μM; ziram 5 μM; b undifferentiated LUHMES cells subjected to the same treatments

Article Snippet: Chemicals Chemicals included Ferbam from TCI America, Portland, OR; ML385 and MS275 from Toris Biosciences, Minneapolis, MN; APTO235 (MedChemExpress, Princeton, NJ); and sankel (Muse Chem, Fairfield, NJ).

Techniques: Expressing, Quantitative RT-PCR

Journal: Frontiers in Pharmacology

Article Title: Melatonin Alleviates Acute Sleep Deprivation-Induced Memory Loss in Mice by Suppressing Hippocampal Ferroptosis

doi: 10.3389/fphar.2021.708645

Figure Lengend Snippet: Effect of melatonin on MT2/ERK/Nrf2 signaling in the hippocampus of sleep-deprived mice (A, C) Immunohistochemical staining of MT2 in hippocampus. Brown indicates positive cells. (B, D) negative control. A–B: bar = 200 μm; C–D: bar = 50 μm. (E–H) Relative protein levels of MT1, MT2, p -ERK1/2 and Nrf2 were normalized to β-actin ( n = 6). Differences were assessed using one-way ANOVA. The result represents the mean ± standard error of the mean. Values not sharing a common superscript letter differ significantly at p < 0.05; those with the same letter do not differ significantly ( p ≥ 0.05). CON: control group, SD: sleep deprivation group, SD + L-Mel: SD + low melatonin (20 mg/kg) supplement group, SD + H-Mel: SD + high melatonin (40 mg/kg) supplement group.

Article Snippet: Cells were plated before drug treatment in a 6- or 96-well plate at 10 6 or 10 4 cells/well, respectively, and cultured in DMEM with 10% FBS (complete medium) for 6 h. The cells were then cultured in DMEM without FBS (basal medium) for 12 h. The ferroptosis inducer Erastin (E7781; Sigma, United States), the ferroptosis inhibitor Fer-1, the MT2-selective Mel receptor antagonist 4P-PDOT (1034; Tocris Bioscience, Bristol, United Kingdom), the ERK inhibitor PD98059 (1213; Tocris Bioscience, Bristol, United Kingdom) and the Nrf2 inhibitor ML385 (SML1833; Sigma, United States) were each dissolved in DMSO.

Techniques: Immunohistochemical staining, Staining, Negative Control, Control

Journal: Frontiers in Pharmacology

Article Title: Melatonin Alleviates Acute Sleep Deprivation-Induced Memory Loss in Mice by Suppressing Hippocampal Ferroptosis

doi: 10.3389/fphar.2021.708645

Figure Lengend Snippet: Effect of melatonin on MT2/ERK/Nrf2 signaling in the HT-22 cells exposed to Erastin. The HT-22 cells exposed to Erastin and melatonin or Fer-1 were pretreated with 4P-PDOT, PD98059 and ML385, respectively. (A) Relative cell viability (%) ( n = 5). (B) Fluorescence staining of ROS. Bar = 200 μm. (C) Relative quantification of intracellular ROS level ( n = 5). (D–J) Relative protein levels of p -ERK1/2, Nrf2, GPX4, TFR1, DMT1 and FPN normalized to β-actin ( n = 6). (K–L) SOD and MDA levels ( n = 6). (M–O) Relative mRNA levels of iron transporter proteins (TFR1, DMT1 and FPN) ( n = 6). Differences were assessed using one-way ANOVA. The result represents the mean ± standard error of the mean. Values not sharing a common superscript letter differ significantly at p < 0.05; those with the same letter do not differ significantly ( p ≥ 0.05).

Article Snippet: Cells were plated before drug treatment in a 6- or 96-well plate at 10 6 or 10 4 cells/well, respectively, and cultured in DMEM with 10% FBS (complete medium) for 6 h. The cells were then cultured in DMEM without FBS (basal medium) for 12 h. The ferroptosis inducer Erastin (E7781; Sigma, United States), the ferroptosis inhibitor Fer-1, the MT2-selective Mel receptor antagonist 4P-PDOT (1034; Tocris Bioscience, Bristol, United Kingdom), the ERK inhibitor PD98059 (1213; Tocris Bioscience, Bristol, United Kingdom) and the Nrf2 inhibitor ML385 (SML1833; Sigma, United States) were each dissolved in DMSO.

Techniques: Fluorescence, Staining, Quantitative Proteomics

Journal: Frontiers in Pharmacology

Article Title: Melatonin Alleviates Acute Sleep Deprivation-Induced Memory Loss in Mice by Suppressing Hippocampal Ferroptosis

doi: 10.3389/fphar.2021.708645

Figure Lengend Snippet: Hypothetical diagram of how melatonin improves SD-induced hippocampal ferroptosis. Exogenous melatonin likely alleviates hippocampal ferroptosis caused by acute SD through by binding to the MT2 receptor and activating ERK/Nrf2 signaling, thereby improving lipid peroxidation and iron transporter disorder. ARE, antioxidant response element; CP, ceruloplasmin; DMT1, divalent metal transporter 1; ERK1/2, extracellular regulated protein kinases; FPN, ferroportin; GPX4, glutathione peroxidase 4; Keap1, Kelch-like ECH-associated protein 1; Mel, melatonin; ML385, Nrf2 inhibitor; MDA, malondialdehyde; MT2, melatonin receptor 2; Nrf2, nuclear factor erythroid 2-related factor 2; PD98059, ERK inhibitor; ROS, reactive oxygen species; SD, sleep deprivation; SOD, superoxide dismutase; STEAP3, 6-transmembrane epithelial antigen of the prostate 3; TFR1, transferrin receptor 1; 4P-PDOT, 4-phenyl-2-propionamidotetralin.

Article Snippet: Cells were plated before drug treatment in a 6- or 96-well plate at 10 6 or 10 4 cells/well, respectively, and cultured in DMEM with 10% FBS (complete medium) for 6 h. The cells were then cultured in DMEM without FBS (basal medium) for 12 h. The ferroptosis inducer Erastin (E7781; Sigma, United States), the ferroptosis inhibitor Fer-1, the MT2-selective Mel receptor antagonist 4P-PDOT (1034; Tocris Bioscience, Bristol, United Kingdom), the ERK inhibitor PD98059 (1213; Tocris Bioscience, Bristol, United Kingdom) and the Nrf2 inhibitor ML385 (SML1833; Sigma, United States) were each dissolved in DMSO.

Techniques: Binding Assay

Journal: Molecular Medicine

Article Title: Artemisinin ameliorates cognitive decline by inhibiting hippocampal neuronal ferroptosis via Nrf2 activation in T2DM mice

doi: 10.1186/s10020-024-00797-9

Figure Lengend Snippet: ML385 or erastin treatment abolishes the inhibitory effect of Art on neuronal ferroptosis in the hippocampus. T2DM mice were coadministered artemisinin (Art, 40 mg/kg/d, i.p.) and ML385 (30 mg/kg, i.p.) or erastin (30 mg/kg, i.p.) for 4 consecutive weeks. A Representative transmission electron microscopy (TEM) images of mitochondrial morphology (normal mitochondria are indicated with red arrows, and shrunken and broken mitochondria are indicated with yellow arrows) in the hippocampal CA1 area of the mice. Scale bar, 500 nm. B – E ROS, MDA, GSH, and Fe 2+ contents in the hippocampal CA1 area were detected using assay kits. The data are expressed as the mean ± SEM (n = 4 per group). * P < 0.05, ** P < 0.001

Article Snippet: Art (purity ≥ 98%), erastin, and ML385 were purchased from APExBIO Technology LLC (Houston, USA).

Techniques: Transmission Assay, Electron Microscopy

Journal: Molecular Medicine

Article Title: Artemisinin ameliorates cognitive decline by inhibiting hippocampal neuronal ferroptosis via Nrf2 activation in T2DM mice

doi: 10.1186/s10020-024-00797-9

Figure Lengend Snippet: Cotreatment with ML385 or erastin abolishes the neuro-protective effect of Art on T2DM mice. T2DM mice were cotreated with artemisinin (Art, 40 mg/kg/d, i.p.) and ML385 (30 mg/kg, i.p.) or erastin (30 mg/kg, i.p.) for 4 consecutive weeks. Neuronal injury and neuron loss in the hippocampus were measured by H&E staining. A Representative images of the hippocampal CA1 region by H&E staining. Scale bar, 100 µm. B Statistical analyses for neuron number in the CA1 region. The data are expressed as the mean ± SEM (n = 4 per group). * P < 0.05

Article Snippet: Art (purity ≥ 98%), erastin, and ML385 were purchased from APExBIO Technology LLC (Houston, USA).

Techniques: Staining

Journal: Molecular Medicine

Article Title: Artemisinin ameliorates cognitive decline by inhibiting hippocampal neuronal ferroptosis via Nrf2 activation in T2DM mice

doi: 10.1186/s10020-024-00797-9

Figure Lengend Snippet: Cotreatment with ML385 or erastin reverses the ameliorative effect of Art on cognitive deficits in T2DM mice. T2DM mice were cotreated with artemisinin (Art, 40 mg/kg/d, i.p.) and ML385 (30 mg/kg, i.p.) or erastin (30 mg/kg, i.p.) for 4 consecutive weeks. A Total entry times and ( B ) the alternation rate were detected in the Y maze test. C The escape latency to find the platform in the navigation phase, D time spent in the target quadrant and ( E ) number of platform crossings during the probe trial phase in the MWM test were also analysed. Data are the mean ± SEM (n = 12 per group). * P < 0.05 vs the T2DM + Art group

Article Snippet: Art (purity ≥ 98%), erastin, and ML385 were purchased from APExBIO Technology LLC (Houston, USA).

Techniques:

Journal: Heliyon

Article Title: Fraxin inhibits melanogenesis by suppressing the ERK/MAPK pathway and antagonizes oxidative stress by activating the NRF2 pathway

doi: 10.1016/j.heliyon.2023.e18929

Figure Lengend Snippet: The role of fraxin on melanogenesis and intracellular ROS in UVB-treated MNT1 cells. (a) MNT1 cells was pretreatment with fraxin for 1 day, then treated with UVB (30 mJ/cm 2 ) and ML385 (20 μM) for 2 days, (a) the melanin content was observed by Fontana-Masson staining, (b) the level of intracellular ROS was observed by DCFH-DA fluorescencent probes, (c) the expression of NRF2, CAT, and HO-1 proteins was measured by western blotting. (d) The schematic diagram of the molecular mechanism.

Article Snippet: ML385 was purchased from Adooq Bioscience.

Techniques: Staining, Expressing, Western Blot