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: CNS Neuroscience & Therapeutics

Article Title: Acetyl‐11‐keto‐beta‐boswellic acid modulates macrophage polarization and Schwann cell migration to accelerate spinal cord injury repair in rats

doi: 10.1111/cns.14642

Figure Lengend Snippet: Information of primary antibodies.

Article Snippet: It has been shown that AKBA can promote the expression of Nrf2, so the group with the addition of Nrf2 inhibitor (ML385) (5 μm) (MedChemExpress, Monmouth Junction, NJ, USA) was designed.

Techniques: Concentration Assay

Journal: CNS Neuroscience & Therapeutics

Article Title: Acetyl‐11‐keto‐beta‐boswellic acid modulates macrophage polarization and Schwann cell migration to accelerate spinal cord injury repair in rats

doi: 10.1111/cns.14642

Figure Lengend Snippet: Primer sequence list.

Article Snippet: It has been shown that AKBA can promote the expression of Nrf2, so the group with the addition of Nrf2 inhibitor (ML385) (5 μm) (MedChemExpress, Monmouth Junction, NJ, USA) was designed.

Techniques: Sequencing

Journal: CNS Neuroscience & Therapeutics

Article Title: Acetyl‐11‐keto‐beta‐boswellic acid modulates macrophage polarization and Schwann cell migration to accelerate spinal cord injury repair in rats

doi: 10.1111/cns.14642

Figure Lengend Snippet: AKBA can activate the expression of Nrf2 pathway to reduce oxidative stress. (A) Pictures of the fluorescent expression of Nrf2 in the spinal cord. The white scale is 20 μm. (B) Quantitative analysis of Nrf2 fluorescence pictures on day 7. (C) Nrf2/HO‐1 protein expression map. (D) Quantitative analysis of Nrf2 protein on day 7. (E) Quantitative analysis of HO‐1 protein on day 7. (F) The mRNA relative expression level of Nrf2 on the 7th day. (G) Relative mRNA expression level of HO‐1 on day 7. (H) CAT content. (I) GSH‐PX content. (J) T‐AOC content. (K) SOD enzyme activity. (L) MDA content. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: It has been shown that AKBA can promote the expression of Nrf2, so the group with the addition of Nrf2 inhibitor (ML385) (5 μm) (MedChemExpress, Monmouth Junction, NJ, USA) was designed.

Techniques: Expressing, Fluorescence, Activity Assay

Journal: CNS Neuroscience & Therapeutics

Article Title: Acetyl‐11‐keto‐beta‐boswellic acid modulates macrophage polarization and Schwann cell migration to accelerate spinal cord injury repair in rats

doi: 10.1111/cns.14642

Figure Lengend Snippet: AKBA can increase the anti‐inflammatory expression of macrophages induced by LPS and reduce the release of ROS. (A) Fluorescent expression map of CD206. The white scale is 20 μm. (B) Fluorescence quantitative analysis diagram of CD206. (C) Protein expression map of ARG‐1 and IL‐10. (D) Protein quantitative analysis chart of ARG‐1. (E) Protein quantitative analysis chart of IL‐10. (F) Fluorescence quantitative analysis of ROS. (G) CD206 flow cytometry results. (H) Expression map of ROS. The white scale is 20 μm. (I) Protein expression map of Nrf2 pathway. (J, K) Protein Quantitative Analysis of Nrf2, HO‐1. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: It has been shown that AKBA can promote the expression of Nrf2, so the group with the addition of Nrf2 inhibitor (ML385) (5 μm) (MedChemExpress, Monmouth Junction, NJ, USA) was designed.

Techniques: Expressing, Fluorescence, Flow Cytometry

Journal: CNS Neuroscience & Therapeutics

Article Title: Acetyl‐11‐keto‐beta‐boswellic acid modulates macrophage polarization and Schwann cell migration to accelerate spinal cord injury repair in rats

doi: 10.1111/cns.14642

Figure Lengend Snippet: Nrf2 inhibitors attenuate the ability of AKBA to reduce the iNOS expression. (A) The fluorescence expression map of iNOS. The white scale is 20 μm. (B) Fluorescence quantitative analysis diagram of iNOS. (C) The expression level of iNOS detected by ELISA. (D) Detection level of IL‐1β detected by ELISA. (E) The detection level of TNF‐α detected by ELISA. (F) Protein expression map of TNF‐α and IL‐1β. (G, H) Protein quantitative analysis of TNF‐α, IL‐1β. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: It has been shown that AKBA can promote the expression of Nrf2, so the group with the addition of Nrf2 inhibitor (ML385) (5 μm) (MedChemExpress, Monmouth Junction, NJ, USA) was designed.

Techniques: Expressing, Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: CNS Neuroscience & Therapeutics

Article Title: Acetyl‐11‐keto‐beta‐boswellic acid modulates macrophage polarization and Schwann cell migration to accelerate spinal cord injury repair in rats

doi: 10.1111/cns.14642

Figure Lengend Snippet: Nrf2 inhibitors attenuate the ability of AKBA to increase the CD206 expression. (A) Fluorescent expression map of CD206. The white scale is 20 μm. (B) Fluorescence quantitative analysis of CD206. (C) Protein expression maps of ARG‐1 and IL‐10. (D, E) Protein quantitative analysis of ARG‐1, IL‐10. (F) Expression map of ROS. The white scale is 50 μm. (G) Protein expression map of Nrf2/HO‐1. (H, I) Protein quantitative analysis of Nrf2 and HO‐1. (J) Quantitative analysis of ROS. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: It has been shown that AKBA can promote the expression of Nrf2, so the group with the addition of Nrf2 inhibitor (ML385) (5 μm) (MedChemExpress, Monmouth Junction, NJ, USA) was designed.

Techniques: Expressing, Fluorescence

Journal: CNS Neuroscience & Therapeutics

Article Title: Acetyl‐11‐keto‐beta‐boswellic acid modulates macrophage polarization and Schwann cell migration to accelerate spinal cord injury repair in rats

doi: 10.1111/cns.14642

Figure Lengend Snippet: Nrf2 inhibitors attenuate the ability of AKBA to promote Schwann cell migration. (A) Detection of S100 at the site of spinal cord injury. (B) AKBA‐treated macrophage culture promotes the migration ability of Schwann cells. (C) Statistical graph of Schwann cell migration. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: It has been shown that AKBA can promote the expression of Nrf2, so the group with the addition of Nrf2 inhibitor (ML385) (5 μm) (MedChemExpress, Monmouth Junction, NJ, USA) was designed.

Techniques: Migration

Journal: CNS Neuroscience & Therapeutics

Article Title: Acetyl‐11‐keto‐beta‐boswellic acid modulates macrophage polarization and Schwann cell migration to accelerate spinal cord injury repair in rats

doi: 10.1111/cns.14642

Figure Lengend Snippet: AKBA promotes rat macrophage polarization and Schwann cell migration. (A) iNOS fluorescence image. Red fluorescence indicates iNOS. (B) iNOS fluorescence quantitative analysis chart. (C) CD206 fluorescence image. Red fluorescence indicates CD206. (D) CD206 fluorescence quantitative analysis chart. (E) CD206 flow chart. (F) CD206 flow cytometry analysis diagram. (G) ROS staining picture. (H) ROS fluorescence analysis chart. (I) The migration ability of rat macrophage culture medium on SC cells in each group. (J) Analysis of Schwann cell migration numbers. (K) Protein expression diagram of TNF‐α and IL‐1β. (L) Protein quantitative analysis chart of IL‐1β. (M) Quantitative analysis chart of TNF‐α. (N) Protein expression diagram of ARG‐1 and IL‐10. (O) ARG‐1 protein quantitative analysis chart. (P) Quantitative protein analysis chart of IL‐10. (Q) Protein expression diagram of Nrf2 and HO‐1. Protein quantitative analysis chart of (R) Nrf2. (S) HO‐1 protein quantitative analysis chart. C represents the blank group, C + M represents the blank plus ML385 group, C + A represents the blank plus AKBA group, and C + A + M represents the blank plus AKBA and ML385. L represents the LPS group, L + A represents the LPS + AKBA group, L + M represents the LPS + ML385 group, and L + A + M represents the LPS + AKBA + ML385 group. The lowercase letters above the statistical graph indicate differences. The same letters indicate no significant difference, and different letters indicate significant differences ( p < 0.05).

Article Snippet: It has been shown that AKBA can promote the expression of Nrf2, so the group with the addition of Nrf2 inhibitor (ML385) (5 μm) (MedChemExpress, Monmouth Junction, NJ, USA) was designed.

Techniques: Migration, Fluorescence, Flow Cytometry, Staining, Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Dexmedetomidine alleviates inflammation in neuropathic pain by suppressing NLRP3 via Nrf2 activation

doi: 10.3892/etm.2021.10479

Figure Lengend Snippet: Experimental schedule. A total of 138 rats were included in the present study. A total of 84 rats underwent behavioral tests on day 0, 1, 3, 7 and 14 after the operation, including the DEX+1, DEX+2 and DEX+5 groups, two sham groups, two model groups and one control, DEX, MCC950, nigericin, DEX + ML385, MCC950 + ML385 and DEX + nigericin groups (n=6 rats per group). A total of 54 rats were sacrificed for spinal cord collection 7 days after the drug administration, including one control, sham, model, DEX, MCC950, nigericin, DEX + ML385, MCC950 + ML385 and DEX + nigericin groups (n=6 rats per group).

Article Snippet: The groups were as follows: Model group (n=6x3; untreated model rats), DEX+1 group (n=6; intraperitoneally injected with 1 μg/kg DEX every day for 7 days after the operation; Jiangsu Nhwa Pharmaceutical Co., Ltd.), DEX+2 (n=6; intraperitoneally injected with 2 μg/kg DEX for 7 days post-operation), DEX+5 group (n=6; intraperitoneally injected with 5 μg/kg DEX for 7 days post-operation), DEX group (n=6x2; intraperitoneally injected with 5 μg/kg DEX for 7 days post-operation), MCC950 group [n=6x2; intraperitoneally injected with 50 μg/kg MCC950 (NLRP3 antagonist) for 7 days post-operation; MedChemExpress], nigericin group [n=6x2; intraperitoneally injected with 1 mg/kg nigericin (NLRP3 activator) for 7 days post-operation; MedChemExpress], DEX + ML385 group [n=6x2; intraperitoneally injected with 5 μg/kg DEX and 30 mg/kg ML385 (Nrf2 inhibitor) for 7 days post-operation; Selleck Chemicals], DEX + nigericin group (n=6x2; intraperitoneally injected with 5 μg/kg DEX and 1 mg/kg nigericin for 7 days post-operation) and MCC950 + ML385 group (n=6x2; intraperitoneally injected with 30 mg/kg ML385 and 50 μg/kg MCC950 for 7 days post-operation).

Techniques: Control

Journal: Experimental and Therapeutic Medicine

Article Title: Dexmedetomidine alleviates inflammation in neuropathic pain by suppressing NLRP3 via Nrf2 activation

doi: 10.3892/etm.2021.10479

Figure Lengend Snippet: DEX reduces neuropathic pain by activating Nrf2 in CCI rats. CCI rats were injected with DEX and ML385. (A) Reverse transcription-quantitative PCR and (B) western blotting were performed to detect the expression of Nrf2. (C) MWT was recorded in response to mechanical stimulation. (D) Hematoxylin and eosin staining revealed spinal cord injury (magnification, x400). (E) TUNEL staining was performed to examine the apoptosis in the spinal cords (magnification, x400). (F) The expression levels of inflammatory factors were measured via ELISA. (G) Western blotting was performed to detect the expression of NLRP3. n=6 per group. * P<0.05; ** P<0.01; *** P<0.001. DEX, dexmedetomidine; CCI, chronic constriction injury; MWT, mechanical withdrawal threshold; Nrf2, nuclear factor erythroid 2-related factor 2; NLRP3, NLR family pyrin domain containing 3; g, pressure unit.

Article Snippet: The groups were as follows: Model group (n=6x3; untreated model rats), DEX+1 group (n=6; intraperitoneally injected with 1 μg/kg DEX every day for 7 days after the operation; Jiangsu Nhwa Pharmaceutical Co., Ltd.), DEX+2 (n=6; intraperitoneally injected with 2 μg/kg DEX for 7 days post-operation), DEX+5 group (n=6; intraperitoneally injected with 5 μg/kg DEX for 7 days post-operation), DEX group (n=6x2; intraperitoneally injected with 5 μg/kg DEX for 7 days post-operation), MCC950 group [n=6x2; intraperitoneally injected with 50 μg/kg MCC950 (NLRP3 antagonist) for 7 days post-operation; MedChemExpress], nigericin group [n=6x2; intraperitoneally injected with 1 mg/kg nigericin (NLRP3 activator) for 7 days post-operation; MedChemExpress], DEX + ML385 group [n=6x2; intraperitoneally injected with 5 μg/kg DEX and 30 mg/kg ML385 (Nrf2 inhibitor) for 7 days post-operation; Selleck Chemicals], DEX + nigericin group (n=6x2; intraperitoneally injected with 5 μg/kg DEX and 1 mg/kg nigericin for 7 days post-operation) and MCC950 + ML385 group (n=6x2; intraperitoneally injected with 30 mg/kg ML385 and 50 μg/kg MCC950 for 7 days post-operation).

Techniques: Injection, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Staining, TUNEL Assay, Enzyme-linked Immunosorbent Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Dexmedetomidine alleviates inflammation in neuropathic pain by suppressing NLRP3 via Nrf2 activation

doi: 10.3892/etm.2021.10479

Figure Lengend Snippet: DEX relieves neuropathic pain in CCI rats by suppressing NLRP3 via activation of Nrf2. CCI rats were injected with MCC950 + ML385 or Nigericin + DEX. (A) Western blotting was performed to detect the expression of NLRP3. (B) MWT was recorded in response to mechanical stimulation. (C) Hematoxylin and eosin staining revealed spinal cord injury (magnification, x400). (D) TUNEL staining was performed to examine the apoptosis in the spinal cords (magnification, x400). (E) The expression levels of inflammatory factors were measured via ELISA. n=6 per group. * P<0.05; ** P<0.01. DEX, dexmedetomidine; CCI, chronic constriction injury; MWT, mechanical withdrawal threshold; Nrf2, nuclear factor erythroid 2-related factor 2; NLRP3, NLR family pyrin domain containing 3; g, pressure unit.

Article Snippet: The groups were as follows: Model group (n=6x3; untreated model rats), DEX+1 group (n=6; intraperitoneally injected with 1 μg/kg DEX every day for 7 days after the operation; Jiangsu Nhwa Pharmaceutical Co., Ltd.), DEX+2 (n=6; intraperitoneally injected with 2 μg/kg DEX for 7 days post-operation), DEX+5 group (n=6; intraperitoneally injected with 5 μg/kg DEX for 7 days post-operation), DEX group (n=6x2; intraperitoneally injected with 5 μg/kg DEX for 7 days post-operation), MCC950 group [n=6x2; intraperitoneally injected with 50 μg/kg MCC950 (NLRP3 antagonist) for 7 days post-operation; MedChemExpress], nigericin group [n=6x2; intraperitoneally injected with 1 mg/kg nigericin (NLRP3 activator) for 7 days post-operation; MedChemExpress], DEX + ML385 group [n=6x2; intraperitoneally injected with 5 μg/kg DEX and 30 mg/kg ML385 (Nrf2 inhibitor) for 7 days post-operation; Selleck Chemicals], DEX + nigericin group (n=6x2; intraperitoneally injected with 5 μg/kg DEX and 1 mg/kg nigericin for 7 days post-operation) and MCC950 + ML385 group (n=6x2; intraperitoneally injected with 30 mg/kg ML385 and 50 μg/kg MCC950 for 7 days post-operation).

Techniques: Activation Assay, Injection, Western Blot, Expressing, Staining, TUNEL Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Oncology

Article Title: SIRT5 Promotes Cisplatin Resistance in Ovarian Cancer by Suppressing DNA Damage in a ROS-Dependent Manner via Regulation of the Nrf2/HO-1 Pathway

doi: 10.3389/fonc.2019.00754

Figure Lengend Snippet: SIRT5 inhibits ROS via positively regulating Nrf2/HO-1 pathway. (A) The mRNA expression of Nrf2 and HO-1 in ovarian cancer and normal tissues (GEPIA). (B) Nrf2 and HO-1 protein levels were higher in SKOV-3 and CAOV-3 than A2780 cells assessed by western blot and the relative gray values were shown in histogram. (C) The correlation between Nrf2 and SIRT5 in ovarian cancer based on GEPIA database by Pearson correlation coefficient. (D) Overexpression of SIRT5 upregulated Nrf2 and HO-1 protein levels and knockdown of SIRT5 reduced their expression assessed by western blot and the relative gray values were shown in histogram. (E,F) Overexpression of SIRT5 facilitated the nuclear translocation of Nrf2 by immunofluorescence staining and western blot after nuclear and cytoplasm isolation. (G) ML385, a specific inhibitor of Nrf2, was added into A2780 cells with terminal concentration of 5 μM for 24 or 48 h and the inhibition efficiency was measured by western blot. (H) The inhibition effect of ROS by overexpression SIRT5 in A2780 cells was reversed when ML385 was applied. (I) Representative flow cytometry results analyzed by FlowJo software. Data are presented as the mean ± SD of three independent experiments. EV, empty vector; NC, negative control; DDP, cisplatin; TPM, transcripts per million. Scale bar = 20 μm, * P < 0.05, ** P <0.01, *** P <0.001, compared with A2780, the EV or NC cells.

Article Snippet: Cisplatin and ML385 (an Nrf2-specific inhibitor) were purchased from MCE, China.

Techniques: Expressing, Western Blot, Over Expression, Knockdown, Translocation Assay, Immunofluorescence, Staining, Isolation, Concentration Assay, Inhibition, Flow Cytometry, Software, Plasmid Preparation, Negative Control

Journal: European journal of histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function.

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Figure 1. Effect of LPS on NRF-2 activation in NRK-52e cells. A) Cell viability after addition of 6–50 μg/mL LPS for 24 h (n=5). B) Cell viability after addition of 50 μg/mL LPS for 6–24 h (n=5). C) Western blots showing NRF2 expression in cells at 6, 12, and 24 h after addition of LPS (50 µg/mL). D) Quantitative analysis of the western blot data, with expression relative to β-actin (n=3). E) Immunofluorescence detection of the expression and distribution of NRF2 following LPS treatment. Data are presented as means ± SD; *p<0.05 vs control.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports.32,33 The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Activation Assay, Western Blot, Expressing, Immunofluorescence, Control

Journal: European journal of histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function.

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Figure 2. Effect of NRF2 inhibition on viability and apoptosis in LPS-induced NRK-52e cells. A) Representative images of apoptotic cells measured by the TUNEL assay. B) Cell viability, determined by the MTT assay (n=5). (C) Western blotting of Bcl-2, Bax, cytochrome c, and NRF2. D-I) Quantitative analysis of the Western blotting data, with expression relative to β-actin, Lamin BI, or COXIV (n=3). Data are presented as the means ± SD; *p<0.05 vs control, #p<0.05 vs LPS.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports.32,33 The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Inhibition, TUNEL Assay, MTT Assay, Western Blot, Expressing, Control

Journal: European journal of histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function.

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Figure 3. Effect of NRF2 on the inflammatory response and oxidative stress in LPS-induced injury of NRK-52e cells. A) Immunoblotting of NRF2 following transfection with LV-NRF2. B) Western blotting of NRF2, Nucleus NRF2, p-p65, IKB-α, p-p38 and erk. C) Quantitative analysis of the Western blotting data, with expression relative to β-actin, p65, p38 or Lamin B1 (n=3). D,E) ELISA of TNF-α and IL-6 in cell culture supernatants (n=5). (F) SOD activity (n=5). G) MDA level (n=5). H) Representative fluores- cence microscopy images of ROS production (DCFH-DA probe). I) Quantitative analysis of the ROS data (n=5). Data are presented as means ± SD; *p<0.05 vs control, #p<0.05 vs LPS.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports.32,33 The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Western Blot, Transfection, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Activity Assay, Microscopy, Control

Journal: European journal of histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function.

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Figure 4. Effect of NRF2 inhibition on viability and apoptosis in LPS-induced NRK-52e cells. A) Representative images of apoptotic cells measured by the TUNEL assay. B) Cell viability, determined by the MTT assay (n=5). (C) Western blotting of Bcl-2, Bax, cytochrome c, and NRF2. D-I) Quantitative analysis of the Western blotting data, with expression relative to β-actin, Lamin BI, or COXIV (n=3). Data are presented as the means ± SD; *p<0.05 vs control, #p<0.05 vs LPS.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports.32,33 The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Inhibition, TUNEL Assay, MTT Assay, Western Blot, Expressing, Control

Journal: European journal of histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function.

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Figure 5. Effect of NRF2 on mitochondrial homeostasis of LPS-induced NRK-52e cells. A) Western blotting of proteins related to mitophagy and mitochondrial-biogenesis. B-F) Quantitative analysis of the Western blotting data, with expression relative to β-actin (n=3). G) Representative fluorescence microscopy images showing co-localization of anti-LC3 II (green)/anti-COXIV (red) antibodies. Data are presented as means ± SD; *p<0.05 vs control, #p<0.05 vs LPS.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports.32,33 The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Western Blot, Expressing, Fluorescence, Microscopy, Control

Journal: European journal of histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function.

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Figure 6. Effect of CLP of rats on renal expression of NRF2. A) Western blotting of NRF2 in kidney tissues. B) Quantitative analysis of the Western blotting data, with expression relative to β-actin (n=5). C) Representative immunohistochemical images showing NRF2 expression. Data are presented as means ±SD; *p<0.05 vs control.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports.32,33 The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Expressing, Western Blot, Immunohistochemical staining, Control

Journal: European journal of histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function.

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Figure 7. Effect of NRF2 activation on oxidative stress, inflammatory responses, and renal function in rats that received CLP. A,B) Western blotting of NRF2 in kidney tissues. C-E) Quantitative analysis Western blotting data, with expression relative to β-actin or Lamin B1 (n=5). F) MDA levels in kidney tissues (n=8). G) SOD activity in kidney tissues (n=8). H,I) ELISA of serum levels TNF-α and IL-6 (n=8). J,K) Serum creatinine and BUN (n=8). L) Representative hematoxylin and eosin staining images of renal tissues. Data are presented as means ± SDs; asterisks, edema; triangle, vacuolization; arrow, loss of brush border; *p<0.05 vs sham; #p<0.05 vs CLP.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports.32,33 The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Activation Assay, Western Blot, Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay, Staining

Journal: European journal of histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function.

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Figure 8. Effect of NRF2 on mitochondrial damage and mitochondrial homeostasis in rats that received CLP. A) Western blotting of kidney proteins related to mitophagy and mitochondrial biogenesis. B-F) Quantitative analysis of the western blotting data with expres- sion relative to β-actin (n=5; data are presented as means ± SD; *p<0.05 vs sham, #p<0.05 vs CLP). G) ATP level in kidney tissues (n=8). H) Representative transmission electron micrographs of renal mitochondria. M, mitochondrion; N, nucleus; CLP, the arrow shows a mitochondrion with swelling and cristae fracture; CLP+ML385, the arrow shows a mitochondrion with swelling, cristae fracture, and rupture of membranes; CLP+TBHQ, the arrow shows a mitochondrion with mild swelling and cristae fracture.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports.32,33 The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Western Blot, Transmission Assay