nrf2 inhibitor  (TargetMol)

Journal: Genes & Diseases

Article Title: TIGAR promotes osteogenic differentiation and ameliorates glucocorticoid-induced osteoporosis via autophagy-Nrf2-ROS axis

doi: 10.1016/j.gendis.2025.101735

Figure Lengend Snippet: TIGAR activated nuclear factor erythroid-2 related factor (Nrf2) to reduce dexamethasone (Dex)-induced oxidative stress through inducing autophagy. Bone marrow mesenchymal stem cells (BMSCs) were transfected with TIGAR overexpression plasmid and treated with Dex. (A, B) Immunofluorescence staining of Nrf2 of BMSCs and the quantification of the integrated optical density (IOD) per field. Scale bars, 50 μm. (C, D) Western blot analysis and quantification of Nrf2 expression in extracted nuclear proteins. (E, F) ROS level under Dex treatment in BMSCs with or without administration of 10 nM Nrf2 inhibitor (ML385) after transfecting with TIGAR overexpression plasmid, and the quantification of IOD per filed. Scale bars, 100 μm. BMSCs were treated with Dex and chloroquine (CQ) (20 μM) after transfecting with TIGAR overexpression plasmid. (G–I) Western blot analysis and quantification of p62 and LC3-II expression under different treatments. (J, K) Representative images of mCherry-GFP-LC3 puncta and number of autophagosomes (yellow) (analyzed by Pearson's correlation). Scale bars, 50 μm. (L–N) Western blot analysis and quantification of Nrf2 and kelch-associated protein 1 (Keap1) expression under different treatments. (O, P) The immunofluorescence staining of Nrf2 in BMSCs and the quantification of the IOD per field. Scale bars, 50 μm. (Q, R) Representative immunofluorescence images of LC3 and Keap1. Pearson's correlation of co-localization is shown in the bar graph format from the three independent experiments analyzed. Scale bars, 50 μm. (S, T) Representative images of ROS and the quantification of the IOD per field. Scale bars, 100 μm. Data are shown as mean ± SEM. n = 3, biologically independent samples. Two-way analysis of variance (ANOVA) with Tukey's multiple comparisons test was used to assess statistical significance. ∗ p < 0.05, ∗∗ p < 0.01. NC, negative control. OE, TIGAR overexpression plasmid.

Article Snippet: Nrf2 inhibitor (10 μM) (ML385, TargetMol Chemicals Inc., USA, CAS846557-71-9) and chloroquine (CQ, TargetMol Chemicals Inc., CAS54-05-7) in 20 μg/mL were administered to cells to explore the underlying mechanisms.

Techniques: Transfection, Over Expression, Plasmid Preparation, Immunofluorescence, Staining, Western Blot, Expressing, Negative Control

Journal: Drug Design, Development and Therapy

Article Title: Vitamin K 2 Protects Against Glucocorticoid-Induced Osteoporosis by Activating the NRF2/FSP1 Pathway to Inhibit Osteoblast Ferroptosis

doi: 10.2147/DDDT.S554610

Figure Lengend Snippet: The mechanism of VK 2 in preventing GIOP through the NRF2/FSP1 pathway to inhibit ferroptosis in osteoblasts.

Article Snippet: Based on specific experimental objectives, cells were additionally treated with the ferroptosis inducer FIN56 (5μm, HY-103087, MCE, China), the FSP1 inhibitor iFSP1 (3μm, HY-136057, MCE, China), or the NRF2 inhibitor ML385 (50μm, HY-100523, MCE, China), RSL3 (800nM, HY-100218A, MCE, China).

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Molecular insights into Atorvastatin’s role in delaying intervertebral disc degeneration

doi: 10.3389/fcell.2025.1693951

Figure Lengend Snippet: Ator promotes Nrf2 expression and its downstream gene activation. Nrf2 expression was detected by pretreatment with Ator for 6 h with the addition of 300 μM H 2 O 2 . (A) Immunofluorescence for Nrf2 expression. (B–G) Western blotting was performed to detect the expression of Total-Nrf2, HO-1, NQO-1, and Cytosolic-Nrf2 with β-actin as an internal reference and Nuclear-Nrf2 with Lamin-b as an internal reference. (H–J) Detection of Nrf2, NQO-1, and HO-1 expression using qPCR. All the data are shown as mean ± deviation, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001. Abbreviations: Ator, atorvastatin; ns, non-significant.

Article Snippet: NPC were divided into five groups: control group (routine culture without any treatment), H 2 O 2 group (induced using 300 um/mL H 2 O 2 ), Ator treatment group (10 umol/mL Ator (MCE, US) and 300 umol/mL H 2 O 2 co-treatment were selected), Nrf2 inhibitor group (10 μM Nrf2-IN-1 (MCE, US) plus 300 umol/mL H 2 O 2 co-treatment), Nrf2 inhibitor plus Ator group (10uM Nrf2-IN-1 plus 300 umol/mL H 2 O 2 and 10 umol/mL Ator co-treatment) Cells were incubated at 37 °C in a 5% CO 2 incubator for subsequent experiments.

Techniques: Expressing, Activation Assay, Immunofluorescence, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: Molecular insights into Atorvastatin’s role in delaying intervertebral disc degeneration

doi: 10.3389/fcell.2025.1693951

Figure Lengend Snippet: Activation of Nrf2 is necessary for Ator to exert antioxidant effects. Addition of Nrf2-IN-1 treatment followed by Ator pretreatment for 6 h and H 2 O 2 treatment for 6 h (A–F) Western blotting for detection of relevant protein expression. Data are shown as mean ± deviation, n = 3. (G–I) qPCR for Nrf2, NQO-1, HO-1 expression. Data are shown as mean ± deviation, n = 3. (J) Antioxidant enzyme SOD expression. (K) ROS expression level. (L) CCK8 assay for cellular activity. Data are shown as mean ± deviation, n = 6. *P < 0.05, **P < 0.01, ***P < 0.001. Abbreviations: Ator, atorvastatin; Nrf2-IN-1, Nrf2 inhibitor; ns, non-significant.

Article Snippet: NPC were divided into five groups: control group (routine culture without any treatment), H 2 O 2 group (induced using 300 um/mL H 2 O 2 ), Ator treatment group (10 umol/mL Ator (MCE, US) and 300 umol/mL H 2 O 2 co-treatment were selected), Nrf2 inhibitor group (10 μM Nrf2-IN-1 (MCE, US) plus 300 umol/mL H 2 O 2 co-treatment), Nrf2 inhibitor plus Ator group (10uM Nrf2-IN-1 plus 300 umol/mL H 2 O 2 and 10 umol/mL Ator co-treatment) Cells were incubated at 37 °C in a 5% CO 2 incubator for subsequent experiments.

Techniques: Activation Assay, Western Blot, Expressing, CCK-8 Assay, Activity Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Molecular insights into Atorvastatin’s role in delaying intervertebral disc degeneration

doi: 10.3389/fcell.2025.1693951

Figure Lengend Snippet: Schematic representation of Ator inhibition of H 2 O 2 -induced apoptosis in NPCs. Ator action on NPC causes Nrf2 to separate from Keap1 and enter the nucleus, which combines with ARE to promote the translation of downstream antioxidant proteins such as HO-1 and NQO-1 and inhibit H 2 O 2 -induced oxidative stress. It also reduced the activation of caspase-3 and inhibited apoptosis in NPC. Abbreviations:Nrf2, Nuclear factor erythroid 2-related factor 2; Keap1, Kelch-like ECH-associated protein 1; ROS, Reactive Oxygen Species; ARE, Antioxidant Response Element; HO-1, Heme Oxygenase-1; NQO-1, NAD(P) H Quinone Oxidoreductase 1; Bcl2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein; H 2 O 2 , Hydrogen Peroxide.

Article Snippet: NPC were divided into five groups: control group (routine culture without any treatment), H 2 O 2 group (induced using 300 um/mL H 2 O 2 ), Ator treatment group (10 umol/mL Ator (MCE, US) and 300 umol/mL H 2 O 2 co-treatment were selected), Nrf2 inhibitor group (10 μM Nrf2-IN-1 (MCE, US) plus 300 umol/mL H 2 O 2 co-treatment), Nrf2 inhibitor plus Ator group (10uM Nrf2-IN-1 plus 300 umol/mL H 2 O 2 and 10 umol/mL Ator co-treatment) Cells were incubated at 37 °C in a 5% CO 2 incubator for subsequent experiments.

Techniques: Inhibition, Activation Assay