nrf2 inhibitor ml385  (TargetMol)

Journal: Bioactive Materials

Article Title: Metformin alleviates aging-associated periodontitis via NRF2-mediated restoration of the IRE1α dependent unfolded protein response

doi: 10.1016/j.bioactmat.2026.03.060

Figure Lengend Snippet: SASP and NRF2 expression in periodontal tissues from young and aged individuals with or without periodontitis. Periodontal tissues and PDLSCs were collected from young (≤25 years), middle-aged (35-45 years), and aged (≥60 years) individuals, with or without periodontitis. (A) Schematic diagram of the experimental procedure. (B, C) SA-β-gal (B) and P21 (C) staining in periodontal tissues of young (Young), young individuals with periodontitis (Young + PD), aged individuals (Aged), and aged individuals with periodontitis (Aged + PD). (D) RT-qPCR analysis of mRNA expression levels of IL1β , TNFα , IL6 , IL8 , MMP3 , and MMP13 in periodontal tissues. (E) IF staining for NFR2 in periodontal tissues. (F) Western blot analysis of NRF2 expression in PDLSCs collected from young (Young), middle-aged (Middle), and aged (Aged) individuals. (G) NRF2 expression levels in PDLSCs at different passages (Passage 3, 9, and 12) with or without LPS (10 μg/ml) treatment. ns, not significant; PD, periodontitis; IF, Immunofluorescence. Data are presented as means ± SEM, with n = 3-5 for each subgroup. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: To assess the role of NRF2, PDLSCs were pretreated with ML385 ( HY100523 , MCE), an NRF2 inhibitor, for 24 h. For NRF2 overexpression, PDLSCs were transduced with an adenoviral vector encoding NRF2 (Adv- NRF2 ; titer: 7.11 × 10 10 ) or a control vector (Adv- GFP ; titer: 8.0 × 10 10 ) (OBiO Inc., Shanghai, China) for 24 h. To knock down IRE1α, PDLSCs were transfected with siRNA targeting IRE1α (sense: 5′-GUUUGAUCCCGGACUCAAATT-3′; antisense: 5′-UUUGAGUCCGGGAUCAAAC TT-3′) or a scrambled control siRNA.

Techniques: Expressing, Staining, Quantitative RT-PCR, Western Blot, Immunofluorescence

Journal: Bioactive Materials

Article Title: Metformin alleviates aging-associated periodontitis via NRF2-mediated restoration of the IRE1α dependent unfolded protein response

doi: 10.1016/j.bioactmat.2026.03.060

Figure Lengend Snippet: Overexpression of Nrf2 suppresses SASP in senescent PDLSCs. PDLSCs were transduced with adenovirus carrying Nrf2 (Adv- NRF2 ) or control virus (Adv- GFP ), followed by treatment with LPS (10 μg/ml) with or without D-gal (30 mg/ml). (A) Representative images of SA-β-gal staining, followed by quantification. (B) Western blot analysis of P16 and P21 protein levels. (C) mRNA expression levels of IL1β , TNFα , IL6 , IL8 , TNFα , MMP3 , and MMP13 in PDLSCs. (D, E) ALP (D) and ARS (E) staining of PDLSCs. (F) Western blot analysis and quantification of COL-1α1, RUNX2, OPN, and ALP protein expression levels in PDLSCs after 7 days of osteogenic induction. ns, not significant. Data are presented as means ± SEM (n = 3 per subgroup). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: To assess the role of NRF2, PDLSCs were pretreated with ML385 ( HY100523 , MCE), an NRF2 inhibitor, for 24 h. For NRF2 overexpression, PDLSCs were transduced with an adenoviral vector encoding NRF2 (Adv- NRF2 ; titer: 7.11 × 10 10 ) or a control vector (Adv- GFP ; titer: 8.0 × 10 10 ) (OBiO Inc., Shanghai, China) for 24 h. To knock down IRE1α, PDLSCs were transfected with siRNA targeting IRE1α (sense: 5′-GUUUGAUCCCGGACUCAAATT-3′; antisense: 5′-UUUGAGUCCGGGAUCAAAC TT-3′) or a scrambled control siRNA.

Techniques: Over Expression, Transduction, Control, Virus, Staining, Western Blot, Expressing

Journal: Bioactive Materials

Article Title: Metformin alleviates aging-associated periodontitis via NRF2-mediated restoration of the IRE1α dependent unfolded protein response

doi: 10.1016/j.bioactmat.2026.03.060

Figure Lengend Snippet: Nrf2 knockout aggravates SASP and alveolar bone resorption. (A) Schematic diagram of the experimental procedure. (B, C) IF staining of SA-β-gal (B) and P21 (C) in periodontal tissues from WT and Nrf2 −/− mice, settled with a periodontitis model and D-gal induced aged mouse model. (D) Relative mRNA level of Il1β in mice periodontal tissue. (E) Micro-CT reconstructions, followed by 3D and 2D views of maxillary molars and evaluation of the distance from the CEJ to the ABC. (F, G) Mice periodontal tissue sections prepared for H&E staining (F) and Masson's trichrome staining (G). ns, not significant. Data are presented as means ± SEM (n = 5 per subgroup). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: To assess the role of NRF2, PDLSCs were pretreated with ML385 ( HY100523 , MCE), an NRF2 inhibitor, for 24 h. For NRF2 overexpression, PDLSCs were transduced with an adenoviral vector encoding NRF2 (Adv- NRF2 ; titer: 7.11 × 10 10 ) or a control vector (Adv- GFP ; titer: 8.0 × 10 10 ) (OBiO Inc., Shanghai, China) for 24 h. To knock down IRE1α, PDLSCs were transfected with siRNA targeting IRE1α (sense: 5′-GUUUGAUCCCGGACUCAAATT-3′; antisense: 5′-UUUGAGUCCGGGAUCAAAC TT-3′) or a scrambled control siRNA.

Techniques: Knock-Out, Staining, Micro-CT

Journal: Bioactive Materials

Article Title: Metformin alleviates aging-associated periodontitis via NRF2-mediated restoration of the IRE1α dependent unfolded protein response

doi: 10.1016/j.bioactmat.2026.03.060

Figure Lengend Snippet: NRF2 restores UPR function through upregulating IRE1α. (A, B) PDLSCs were transduced with an adenovirus vector or loaded with NRF2 (Adv- GFP / NRF2 ), followed by administrated with LPS and/or D-gal. Western blot for UPR markers (A) in PDLSCs. IF staining for IRE1α in PDLSCs (B). (C, D) PDLSCs were administered with NRF2 inhibitor (ML385), followed by treatment with LPS and/or D-gal. Protein levels of UPR markers (C). IF staining for IRE1α (D). (E) IF staining of IRE1α in WT and Nrf2 −/− mice, settled with the periodontitis model and D-gal induced aged mouse model. (F) Prediction of NRF2's binding motifs on the promoters of UPR markers in the JASPAR database ( https://jaspar.elixir.no/ ). (G) Prediction of the relationship between NRF2 and IRE1α mRNA expression in the GEPIA database ( http://gepia.cancer-pku.cn/ ). (H) CUT-RUN-qPCR assay and agarose gel electrophoresis for the binding of NRF2 to IRE1α promoter in PDLSCs. Data are presented as means ± SEM (n = 3 per subgroup). ∗P < 0.05.

Article Snippet: To assess the role of NRF2, PDLSCs were pretreated with ML385 ( HY100523 , MCE), an NRF2 inhibitor, for 24 h. For NRF2 overexpression, PDLSCs were transduced with an adenoviral vector encoding NRF2 (Adv- NRF2 ; titer: 7.11 × 10 10 ) or a control vector (Adv- GFP ; titer: 8.0 × 10 10 ) (OBiO Inc., Shanghai, China) for 24 h. To knock down IRE1α, PDLSCs were transfected with siRNA targeting IRE1α (sense: 5′-GUUUGAUCCCGGACUCAAATT-3′; antisense: 5′-UUUGAGUCCGGGAUCAAAC TT-3′) or a scrambled control siRNA.

Techniques: Transduction, Plasmid Preparation, Western Blot, Staining, Binding Assay, Expressing, Agarose Gel Electrophoresis

Journal: Bioactive Materials

Article Title: Metformin alleviates aging-associated periodontitis via NRF2-mediated restoration of the IRE1α dependent unfolded protein response

doi: 10.1016/j.bioactmat.2026.03.060

Figure Lengend Snippet: Metformin suppresses SASP and upregulates NRF2 and UPR markers expression in senescent PDLSCs. (A) P16 and P21 expression in PDLSCs treated with metformin combined with LPS and D-gal. (B) SA-β-gal staining of PDLSCs. (C) mRNA expression level in PDLSCs. (D) Western blot for NRF2 and UPR markers in PDLSCs treated with metformin combined with LPS and D-gal. (E, F) IF staining for IRE1α (E) and NRF2 (F) in PDLSCs. (G, H) ALP (G) and ARS (H) staining of PDLSCs followed by osteogenic induction for 7 and 21 days, respectively. (I) Protein expression levels of osteogenic markers in PDLSCs were followed by 7 days of osteogenic induction. Data are presented as means ± SEM (n = 3 per subgroup). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: To assess the role of NRF2, PDLSCs were pretreated with ML385 ( HY100523 , MCE), an NRF2 inhibitor, for 24 h. For NRF2 overexpression, PDLSCs were transduced with an adenoviral vector encoding NRF2 (Adv- NRF2 ; titer: 7.11 × 10 10 ) or a control vector (Adv- GFP ; titer: 8.0 × 10 10 ) (OBiO Inc., Shanghai, China) for 24 h. To knock down IRE1α, PDLSCs were transfected with siRNA targeting IRE1α (sense: 5′-GUUUGAUCCCGGACUCAAATT-3′; antisense: 5′-UUUGAGUCCGGGAUCAAAC TT-3′) or a scrambled control siRNA.

Techniques: Expressing, Staining, Western Blot

Journal: Bioactive Materials

Article Title: Metformin alleviates aging-associated periodontitis via NRF2-mediated restoration of the IRE1α dependent unfolded protein response

doi: 10.1016/j.bioactmat.2026.03.060

Figure Lengend Snippet: Metformin suppresses SASP and UPR via NRF2. (A) Schematic diagram of the experimental procedure. PDLSCs were pretreated with D-gal and LPS, followed by administration with metformin and/or NRF2 inhibitor (ML385). (B) Representative images of SA-β-gal staining for PDLSCs. (C) Protein expression level of P21 and P16. (D) Relative mRNA expression of IL1β , IL6 , IL8 , TNFα , MMP3 , and MMP13 in PDLSCs. (E) Western blot analysis of UPR markers expression levels in PDLSCs. (F) Transmission electron microscopy (TEM) images of ER morphology in PDLSCs. ns, not significant. Data are presented as means ± SEM (n = 3 per subgroup). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: To assess the role of NRF2, PDLSCs were pretreated with ML385 ( HY100523 , MCE), an NRF2 inhibitor, for 24 h. For NRF2 overexpression, PDLSCs were transduced with an adenoviral vector encoding NRF2 (Adv- NRF2 ; titer: 7.11 × 10 10 ) or a control vector (Adv- GFP ; titer: 8.0 × 10 10 ) (OBiO Inc., Shanghai, China) for 24 h. To knock down IRE1α, PDLSCs were transfected with siRNA targeting IRE1α (sense: 5′-GUUUGAUCCCGGACUCAAATT-3′; antisense: 5′-UUUGAGUCCGGGAUCAAAC TT-3′) or a scrambled control siRNA.

Techniques: Staining, Expressing, Western Blot, Transmission Assay, Electron Microscopy

Journal: Bioactive Materials

Article Title: Metformin alleviates aging-associated periodontitis via NRF2-mediated restoration of the IRE1α dependent unfolded protein response

doi: 10.1016/j.bioactmat.2026.03.060

Figure Lengend Snippet: TAM-GM@Met suppresses SASP and upregulates NRF2 and UPR markers expression in senescent PDLSCs. (A) Schematic diagram of the co-culture of hydrogels with PDLSCs. (B) Western blot for P16 and P21 protein expression in PDLSCs treated with composite hydrogels combined with LPS and D-gal pre-treated with D-gal and LPS. (C) SA-β-gal staining of PDLSCs. (D) SASP mRNA expression level in PDLSCs. (E, F) ALP (E) and ARS (F) staining of PDLSCs followed by osteogenic induction for 7 and 21 days, respectively. (G) Protein expression levels of osteogenic markers in PDLSCs followed by 7 days of osteogenic induction. (H) Western blot for NRF2 and UPR markers in PDLSCs treated with metformin combined with LPS and D-gal. ns, not significant. Data are presented as means ± SEM (n = 3 per subgroup). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: To assess the role of NRF2, PDLSCs were pretreated with ML385 ( HY100523 , MCE), an NRF2 inhibitor, for 24 h. For NRF2 overexpression, PDLSCs were transduced with an adenoviral vector encoding NRF2 (Adv- NRF2 ; titer: 7.11 × 10 10 ) or a control vector (Adv- GFP ; titer: 8.0 × 10 10 ) (OBiO Inc., Shanghai, China) for 24 h. To knock down IRE1α, PDLSCs were transfected with siRNA targeting IRE1α (sense: 5′-GUUUGAUCCCGGACUCAAATT-3′; antisense: 5′-UUUGAGUCCGGGAUCAAAC TT-3′) or a scrambled control siRNA.

Techniques: Expressing, Co-Culture Assay, Western Blot, Staining

Journal: Bioactive Materials

Article Title: Metformin alleviates aging-associated periodontitis via NRF2-mediated restoration of the IRE1α dependent unfolded protein response

doi: 10.1016/j.bioactmat.2026.03.060

Figure Lengend Snippet: In vivo therapeutic effect of the TAM-GM@Met on mice with an aging-associated periodontitis model. (A) Schematic illustration of the therapeutic process on the D-gal induced aging mice periodontitis model. (B) Micro-CT reconstructions and buccal-palatal sectional views of maxillary molars and evaluation of the distance from the CEJ to the ABC. (C, D) Mice periodontal tissue sections prepared for H&E staining (C) and Masson's trichrome staining (D). (E) Schematic of TAM-GM@Met preparation. TAM hydrogel incorporates galactose-coated MPDA nanoparticles (GM) to target senescent cells for metformin delivery. Metformin activates NRF2, leading to transcriptional upregulation of IRE1α, restoration of the UPR, and suppression of SASP in PDLSCs. ns, not significant; TAM, tannic acid and Ag-MOFs based hydrogel; MPDA, mesoporous polydopamine; UPR, unfolded protein response. Data are presented as means ± SEM (n = 5 per subgroup). ∗∗∗P < 0.001.

Article Snippet: To assess the role of NRF2, PDLSCs were pretreated with ML385 ( HY100523 , MCE), an NRF2 inhibitor, for 24 h. For NRF2 overexpression, PDLSCs were transduced with an adenoviral vector encoding NRF2 (Adv- NRF2 ; titer: 7.11 × 10 10 ) or a control vector (Adv- GFP ; titer: 8.0 × 10 10 ) (OBiO Inc., Shanghai, China) for 24 h. To knock down IRE1α, PDLSCs were transfected with siRNA targeting IRE1α (sense: 5′-GUUUGAUCCCGGACUCAAATT-3′; antisense: 5′-UUUGAGUCCGGGAUCAAAC TT-3′) or a scrambled control siRNA.

Techniques: In Vivo, Micro-CT, Staining

Journal: Renal Failure

Article Title: Curcumin regulates ferroptosis by activating α7 nicotinic acetylcholine receptor to improve sepsis-induced acute kidney injury

doi: 10.1080/0886022X.2026.2657652

Figure Lengend Snippet: LPS induces ferroptosis and inhibits α7nAChR expression in HK-2 cells . (A) HK-2 cells were exposed to varying concentrations of LPS (1, 5, and 10 μg/mL) for 24 h, and the cell viability was subsequently assessed using the CCK-8 assay. (B) HK-2 cells were treated with 5 μg/mL LPS for 12, 24, 48 h, and the cell viability was assessed using the CCK-8. HK-2 cells were exposed to 5 μg/mL LPS for 24 h. (C,D) ELISA was employed to quantify the levels of LDH, TNF-α, and IL-1β in the supernatant of HK-2 cells. (E) Flow cytometry analysis was conducted to measure the ROS level within the cells. (F,G) Biochemical assays were utilized to determine the levels of MDA, GSH, and Fe 2+ in the cells. (H) Western blotting analysis was performed to evaluate the protein expression levels of GPX4, SLC7A11, and NRF2. (I) Immunofluorescence staining was applied to visualize the expression and distribution of α7nAChR protein within the cells. Scale bar = 20 μm. (J) Western blotting analysis was performed to assess the protein expression level of α7nAChR. n = 3. Con, the control group. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Two hours prior to LPS exposure, HK-2 cells were treated with 10 μM NRF2 inhibitor ML385 [ ] (MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Expressing, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Western Blot, Immunofluorescence, Staining, Control

Journal: Renal Failure

Article Title: Curcumin regulates ferroptosis by activating α7 nicotinic acetylcholine receptor to improve sepsis-induced acute kidney injury

doi: 10.1080/0886022X.2026.2657652

Figure Lengend Snippet: Curcumin mitigates LPS-induced injury and ferroptosis in HK-2 cells . (A) HK-2 cells were exposed to varying concentrations of curcumin (2, 5, 10, 20, 50 μmol/L) for 24 h, and the cell viability was subsequently assessed using the CCK-8 assay. (B) The impact of varying concentrations of curcumin (2, 5, 10, 20, 50 μmol/L) on the viability of HK-2 cells exposed to LPS was assessed using the CCK-8 assay. Pre-treat HK-2 cells with 20 μmol/L curcumin for 1 h, followed by co-treatment with 5 μg/mL LPS for 24 h. (C-D) ELISA was employed to quantify the levels of LDH, TNF-α, and IL-1β in the supernatant of HK-2 cells. (E) Biochemical assays were utilized to determine the levels of MDA, GSH, and Fe 2+ in the cells. (F) Flow cytometry analysis was conducted to measure ROS level within the cells. (G,H) qRT-PCR and Western blotting analysis were performed to evaluate the mRNA and protein expression levels of GPX4, SLC7A11, and NRF2. (I) Immunofluorescence staining was applied to visualize the expression and distribution of α7nAChR protein within the cells. Scale bar = 20 μm. (J) Western blotting analysis was performed to assess the protein expression level of α7nAChR. n = 3. Con: the control group; Cur: curcumin. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Two hours prior to LPS exposure, HK-2 cells were treated with 10 μM NRF2 inhibitor ML385 [ ] (MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Quantitative RT-PCR, Western Blot, Expressing, Immunofluorescence, Staining, Control

Journal: Renal Failure

Article Title: Curcumin regulates ferroptosis by activating α7 nicotinic acetylcholine receptor to improve sepsis-induced acute kidney injury

doi: 10.1080/0886022X.2026.2657652

Figure Lengend Snippet: Downregulation of α7nAChR partially reversed the inhibitory effect of curcumin on ferroptosis in LPS-induced HK-2 cells . (A,B) After transfection with si-α7nAChR, the mRNA and protein expression levels of α7nAChR in the cells were detected using qRT-PCR and Western blotting analysis. After transfection with si-α7nAChR, the cells were treated with curcumin and LPS for 24 h. (C) Western blotting analysis was performed to detect the expression level of α7nAChR protein in HK-2 cells. (D) Immunofluorescence staining was utilized to examine the expression and distribution of α7nAChR protein in HK-2 cells. Scale bar = 20 μm. (E,F) Flow cytometry was performed to quantify the production of ROS in HK-2 cells. (G,H) Biochemical assays were employed to measure the levels of MDA, Fe 2+ , and GSH in HK-2 cells. (I,J) qRT-PCR and Western blotting analysis were utilized to assess the mRNA and protein expression levels of GPX4, SLC7A11, and NRF2 in HK-2 cells. n = 3. Con: the control group. Cur: curcumin. ** p < 0.01, *** p < 0.001.

Article Snippet: Two hours prior to LPS exposure, HK-2 cells were treated with 10 μM NRF2 inhibitor ML385 [ ] (MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Transfection, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, Flow Cytometry, Control

Journal: Renal Failure

Article Title: Curcumin regulates ferroptosis by activating α7 nicotinic acetylcholine receptor to improve sepsis-induced acute kidney injury

doi: 10.1080/0886022X.2026.2657652

Figure Lengend Snippet: Curcumin suppresses ferroptosis in renal tissues of CLP-induced SA-AKI mice . (A,B) The levels of MDA, GSH, and Fe 2+ in the renal tissue of SA-AKI mice were measured using biochemical methods. (C) The ROS level in kidney tissue from SA-AKI mice was assessed using DHE staining. Scale bar = 20 μm. (D,E) The mRNA and protein expression levels of GPX4, SLC7A11, and NRF2 in kidney tissues of SA-AKI mice were analyzed by qRT-PCR and Western blotting analysis. n = 6. Cur: curcumin. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Two hours prior to LPS exposure, HK-2 cells were treated with 10 μM NRF2 inhibitor ML385 [ ] (MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Staining, Expressing, Quantitative RT-PCR, Western Blot

Journal: Renal Failure

Article Title: Curcumin regulates ferroptosis by activating α7 nicotinic acetylcholine receptor to improve sepsis-induced acute kidney injury

doi: 10.1080/0886022X.2026.2657652

Figure Lengend Snippet: α7nAChR inhibits ferroptosis in LPS-induced HK-2 cells via NRF2-mediated antioxidant capacity . (A,B) After transfection with Lv-α7nAChR or Lv-NC, the mRNA and protein expression levels of α7nAChR in the cells were detected using qRT-PCR and Western blotting analysis. (C) Flow cytometry was performed to quantify the production of ROS in HK-2 cells. (D,E) Biochemical assays were employed to measure the levels of MDA, GSH, and Fe 2+ in HK-2 cells. (F) The protein expression levels of GPX4, SLC7A11, and NRF2 in HK-2 cells were assessed using Western blotting analysis. n = 3. Con: the control group. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Two hours prior to LPS exposure, HK-2 cells were treated with 10 μM NRF2 inhibitor ML385 [ ] (MedChemExpress, Monmouth Junction, NJ, USA).

Techniques: Transfection, Expressing, Quantitative RT-PCR, Western Blot, Flow Cytometry, Control

Journal: Frontiers in Immunology

Article Title: Fumarate inhibits the formation of neutrophil extracellular traps (NETs) in a Nrf2-controlled and Annexin-A1-dependent manner associated with mitochondrial fusion

doi: 10.3389/fimmu.2026.1770063

Figure Lengend Snippet: Expression of Nrf2 and ANXA1 in relation to NET release by neutrophils treated with dimethyl fumarate. Neutrophils isolated from mice were either pre-treated with DMF (25 µM), ML385 (Nrf2 inhibitor), WRW4 (Fpr2 antagonist) for 1 h or their combination: DMF→ML385, WRW4→DMF incubated with lipopolysaccharide (LPS) at a concentration of 50 μg/mL (for 6 h) or left unstimulated (CTR). (A) Quantification of Nrf2 expression and (B) NETs (extDNA and citH3) quantification after Nrf2 inhibition. (Ci) Representative images and (Cii) area of NETs formed upon the above treatments with WRW4 and DMF. ExtDNA is shown in green and citH3 is shown in red. To visualize co-localization of extDNA with citH3 expression, the images from each channel were overlaid. (D) Levels of ANXA1 were measured in neutrophils supernatants upon the above treatments with DMF and ML385 by ELISA. The results are expressed as the mean values ± SD; n≥3. Values significantly different between the groups (p < 0.05) according to one-way ANOVA ( post hoc Bonferroni test) are designated by letters, where the same letter indicates no differences between groups (different letters indicate statistical differences). Explanation of abbreviations: Nrf2, nuclear factor erythroid 2-related factor 2; ANXA1, Annexin A1; extDNA, extracellular DNA; citH3, citrullinated histone H3.

Article Snippet: The following inhibitors were used: nuclear factor erythroid 2-related factor 2 (NRF2) inhibitor – ML385 (10 μM, MedChemExpress), formyl peptide receptor 2 (Fpr2) antagonist - WRW4 (10 μM, MedChemExpress), NADPH oxidase (NOX) inhibitor - Diphenyleneiodonium chloride (DPI, 10 μM, Merck), necroptosis inhibitor - Necrostatin-1 (50 μM, Torcis Bioscence, Bristol, UK), pyroptosis inhibitor - Disulfiram (30 μM, MedChemExpress) and mitochondrial complex I inhibitor - Rotenone (5 μM, Sigma-Aldrich, Saint Louis, MO, USA).

Techniques: Expressing, Isolation, Incubation, Concentration Assay, Inhibition, Enzyme-linked Immunosorbent Assay