Journal: Life

Article Title: Modulation of Oxidative and ER Stress Pathways by the ADAM17 Inhibitor GW280264X in LPS-Induced Acute Liver Injury

doi: 10.3390/life15121877

Figure Lengend Snippet: Effects of GW280264X on redox regulatory proteins in liver tissue. ( A ) Kelch-like ECH-associated protein 1 (Keap1), ( B ) nuclear factor erythroid 2 2-related factor 2 (Nrf2), and ( C ) glutathione (GSH) levels. Data are presented as mean ± SEM ( n = 5). * p < 0.05 vs. 0.9% NaCl, # p < 0.05 vs. LPS. Black squares represent individual data points for each animal.

Article Snippet: Commercial ELISA kits were used to quantify liver tissue levels of KEAP1 (Cat. No. E2198Mo, BT-Laboratory, Shanghai, China), CHOP (Cat. No. E2718Mo, BT-Laboratory), 4-HNE (Cat. No. E1293Mo, BT-Laboratory), MDA (Cat. No. E0625Mo, BT-Laboratory), NRF2 (Cat. No. E1367Mo, BT-Laboratory), TNF-α (Cat. No. E-EL-M3063, Elabscience, Houston, TX, USA), GRP78 (Cat. No. E-EL-M2696, Elabscience, Houston, TX, USA), glutathione (GSH) colorimetric assay kit (Cat No. E-EL-0026, Elabscience, Houston, TX, USA) and ATF6 (Cat. No. ELK4479, Elk Biotechnology, Wuhan, China).

Techniques:

Journal: Scientific Reports

Article Title: Curcumin preserves bone health compromised by diabetes by inhibiting osteoporosis through regulation of the SIRT3/FoxO3a signalling pathway

doi: 10.1038/s41598-025-15165-8

Figure Lengend Snippet: Curcumin upregulates the expression of the oxidative stress-related proteins Nrf2 and SIRT3 in bone tissue cells. Histograms indicate the SIRT3 protein expression levels relative to the control group. Values are expressed as mean ± standard deviation for three independent experiments ( n = 3). * p < 0.05, *** p < 0.001 vs. control group, # p < 0.05, ## p < 0.01 vs. HG group.

Article Snippet: Curcumin (Solarbio), mouse anti-SIRT3 antibody (Proteintech) , mouse anti-FoxO3a antibody (Proteintech), mouse anti-cytochrome c antibody (Proteintech), mouse anti-NRF2 antibody (Proteintech), mouse anti-NOX2 antibody (Proteintech), mouse anti-caspase 3 antibody (Proteintech), mouse anti-SOD2 antibody (Proteintech), rabbit anti-mouse SIRT3 antibody (Proteintech) Proteintech), rabbit anti-mouse Bcl2 antibody (Proteintech), mouse anti-β-actin antibody (Beijing Zhongyi Jinqiao Company), sodium pentobarbital (Sigma), paraformaldehyde (Xi’an Reagent Factory), α-D-glucose (Sigma), anti-osteocalcin (OCN) antibody (Merck & Millipore ), NQO1 (Proteintech), mouse osteoprotegerin (OPG) antibody (R&D), minimum essential medium alpha (α-MEM) (Bioind), dimethyl sulfoxide Hybri-Max (Sigma), Flow Cytometry Kit (Jiangsu Biyuntian), osteogenic induction complete medium (Pythonbio), Mitochondrial Membrane Potential Assay Kit (Jiangsu Biyuntian), ROS Reactive Oxygen Species Assay Kit (Jiangsu Biyuntian).

Techniques: Expressing, Control, Standard Deviation

Journal: iScience

Article Title: Endothelial cell Nrf2 controls neuroinflammation following a systemic insult

doi: 10.1016/j.isci.2025.112630

Figure Lengend Snippet: Nrf2 activator RTA-404 promotes brain endothelial barrier strength (A) Human BEC cultures were treated with ±RTA-404 and transepithelial/transendothelial electrical resistance (TEER) was measured for 20 h ∗ p = 0.0136, one-way ANOVA plus Sidak post hoc (n = 8–9), data are represented as mean ± SE. (B) Human BEC cultures were treated with ±RTA-404 for 24h, RNA was extracted and RNA-seq was performed. A scatterplot was generated for genes with average expression >0.1 FPKM across the datasets. Highlighted with red and blue crosses are the genes whose expression are significantly increased or decreased respectively ( DESeq2 P _adj < 0.05, n = 3). ‘n’ refers here and throughout as an independent biological replicate (i.e., an independent culture or a mouse). (C) IPA analysis identifying activated pathways in human BEC culture treated with RTA-404 (RTA-404 vs. DMSO Control). Significant DEGs ( DESeq2 P _adj < 0.05, ∣Log 2 FC∣>1, n = 3) with BEC transcriptome as reference dataset were analyzed to calculate the p -value of overlap and Z score of overall activation/inhibition states of individual pathways. The significant activated ( p < 0.05, Z score >2) pathways are shown in the bar chart. (D) Immunocytochemistry showing RTA-404 activates Nrf2 nuclear translocation and upregulates the protein expression of Nrf2 target genes HMOX1 and NQO1 , as well as the gene encoding tight junction protein ZO1. From left to right p = 0.0008, 0.0005, 0.0002, and 0.05, n = 3–4, unpaired t-test. Scale bar = 100 μm. Data are represented as mean ± SE.

Article Snippet: Nrf2 flox mouse , The Jackson Laboratory , Cat. #25433.

Techniques: RNA Sequencing, Generated, Expressing, Control, Activation Assay, Inhibition, Immunocytochemistry, Translocation Assay

Journal: iScience

Article Title: Endothelial cell Nrf2 controls neuroinflammation following a systemic insult

doi: 10.1016/j.isci.2025.112630

Figure Lengend Snippet: Endothelial cell-specific knockout of Nrf2 impairs BEC homeostasis and reduces endothelial barrier strength (A) qRT-PCR confirms gene expression of exon 5 of Nrf2 is specifically deleted in BECs, but not in astrocytes, microglia or oligodendrocytes in Nfe2l2 ENDO mice. Data are represented as mean ± SE, from left to right p = 0.0007, 0.0995, 0.6903, and 0.7432, n = 4, unpaired t-test. (B) Mouse BECs isolated from Nfe2l2 ENDO mice and Nfe2l2 Flox mice were cultured in vitro and TEER was measured for 24hrs. ∗ p = 0.0005, One-way ANOVA plus Sidak post hoc (n = 8–9). Data are represented as mean ± SE. (C) Scatterplot of RNA-seq analysis showing the influence of Nrf2 specific KO in ECs on BEC transcriptome in basal conditions. Two weeks after tamoxifen-induced recombination, BECs were FACS-sorted from Nfe2l2 ENDO mice and their Nfe2l2 fl/fl littermate’s control mice. Scatterplot was generated for genes with average expression >0.1 FPKM across the datasets. Highlighted with red and blue crosses are the genes whose expression are significantly increased or decreased respectively ( DESeq2 P _adj < 0.05, n = 4–6). (D) Volcano plot of mass spectrometry analysis showing the influence of Nrf2 specific KO in ECs on BEC proteome in basal conditions. Two weeks after tamoxifen-induced recombination, BECs were FACS-sorted from Nfe2l2 ENDO mice and their Nfe2l2 fl/fl littermate’s control mice. Highlighted with red and blue dots are the proteins whose expression are significantly increased or decreased respectively ( p < 0.05, n = 4).

Article Snippet: Nrf2 flox mouse , The Jackson Laboratory , Cat. #25433.

Techniques: Knock-Out, Quantitative RT-PCR, Gene Expression, Isolation, Cell Culture, In Vitro, RNA Sequencing, Control, Generated, Expressing, Mass Spectrometry

Journal: iScience

Article Title: Endothelial cell Nrf2 controls neuroinflammation following a systemic insult

doi: 10.1016/j.isci.2025.112630

Figure Lengend Snippet: Nrf2 activator RTA-404 modifies the BEC transcriptome under inflammatory conditions Mice were given i.p. injection of either saline or RTA-404 (n = 6–8) daily for 4 days, following which either saline or LPS was given for 24 h. The brain cells were isolated and then sorted with FACS for BECs, RNA extracted and RNA-seq performed. Scatterplots were generated for genes with average expression >0.1 FPKM across the datasets. Highlighted with red and blue crosses are the genes whose expression are significantly increased or decreased respectively ( DESeq2 P _adj < 0.05, n = 6–8). (A) Scatterplot of RNA-seq analysis showing the BEC transcriptome modified by peripheral LPS insult compared with saline in Nfe2l2 fl/fl mice (LPS vs. Saline). (B) Scatterplot of RNA-seq analysis showing the LPS-induced BEC transcriptome modified by pre-administration of RTA-404 in Nfe2l2 fl/fl mice (LPS+RTA-404 vs. LPS+saline). (C) IPA analysis identifying activated or inhibited pathways by pre-administration of RTA-404 under conditions of LPS exposure (LPS+RTA-404 vs. LPS+saline). Significant DEGs ( DESeq2 P _adj < 0.05, ∣Log 2 FC∣>1, n = 6–8) with the BEC transcriptome as reference dataset were analyzed to calculate the p-value of overlap and Z score of overall activation/inhibition states of individual pathways. The significant activated (p < 0.05, Z score >2) or inhibited (p < 0.05, Z score<−2) pathways are shown in the bar chart in orange and blue respectively. (D) Volcano plot of mass spectrometry analysis showing the LPS-induced BEC proteome modified by pre-administration of RTA-404 in Nfe2l2 fl/fl mice (LPS+RTA-404 vs. LPS+saline). (E) Scatterplot of RNA-seq analysis showing the LPS-induced BEC transcriptome modified by pre-administration of RTA-404 in Nfe2l2 ENDO mice.

Article Snippet: Nrf2 flox mouse , The Jackson Laboratory , Cat. #25433.

Techniques: Injection, Saline, Isolation, RNA Sequencing, Generated, Expressing, Modification, Activation Assay, Inhibition, Mass Spectrometry

Journal: iScience

Article Title: Endothelial cell Nrf2 controls neuroinflammation following a systemic insult

doi: 10.1016/j.isci.2025.112630

Figure Lengend Snippet: Endothelial cell Nrf2 maintains microglial homeostatic signature and regulates microglial activation under inflammatory conditions Mice were given i.p. injection of either saline or RTA-404 (n = 6–8) daily for 4 days, following which either saline or LPS was given for 24 h. The brain cells were isolated and then sorted with FACS for microglia, RNA extracted and RNA-seq performed. Scatterplots were generated for genes with average expression >0.1 FPKM across the datasets. Highlighted with red and blue crosses are the genes whose expression are significantly increased or decreased respectively ( DESeq2 P _adj < 0.05, n = 6–8). (A) Scatterplot of RNA-seq analysis showing the microglia transcriptome modified by peripheral LPS insult compared with saline in Nfe2l2 fl/fl mice (LPS vs. Saline). (B) Scatterplot of RNA-seq analysis showing the LPS-induced microglia transcriptome modified by pre-administration of RTA-404 in Nfe2l2 fl/fl mice (LPS+RTA-404 vs. LPS+saline). (C) Scatterplot of RNA-seq analysis showing the LPS-induced microglia transcriptome modified by pre-administration of RTA-404 in Nfe2l2 ENDO mice. (D) RTA-404 promotes microglial homeostatic signature under inflammatory conditions. Genes considered are those expressed >0.5 FPKM in our data and within the group of microglia signature genes defined by (Butovsky et al., 2014). For each gene, log 2 fold change (log 2 FC) of microglia gene expression in Nfe2l2 fl/fl mice by pre-administration of RTA-404 under conditions of LPS exposure (LPS+RTA-404 vs. LPS+saline) is shown. The data were mined from the complete set shown in (B). ∗P < 0.0001, F(1, 498) = 95.1 relates to main effect of pre-administration of RTA-404 on microglia signature gene expression under inflammatory conditions, two-way ANOVA. (E) IPA analysis identifying activated or inhibited pathways in microglia by pre-administration of RTA-404 under conditions of LPS exposure (LPS+RTA-404 vs. LPS+saline). Significantly DEGs ( DESeq2 P_adj < 0.05, ∣Log 2 FC∣>1, n = 6–8) with the microglia transcriptome as reference dataset were analyzed to calculate the p-value of overlap and Z score of overall activation/inhibition states of individual pathways. The significant activated (p < 0.05, Z score >2) or inhibited (p < 0.05, Z score <-2) pathways are shown in the bar chart in orange and blue respectively. (F) The influence of RTA-404 on LPS-induced genes in microglia. For LPS-induced genes (x axis, FPKM>1 , DESeqP _adj < 0.05, Log 2 FC > 1) in microglia in Nfe2l2 fl/fl mice, the Log 2 FC (y axis) of each gene following treatment with LPS ± RTA-404 in microglia in either Nfe2l2 fl/fl mice or Nfe2l2 ENDO mice were plotted. (iii) ∗p = 0.0014, F (1,6720) = 10.24 relates to main effect of RTA-404 on microglial gene sets induced by LPS, two-way ANOVA. (G) The influence of RTA-404 on LPS-repressed genes in microglia. For LPS-repressed genes (x axis, FPKM>1 , DESeqP _adj < 0.05, Log 2 FC < -1) in microglia in Nfe2l2 fl/fl mice, the Log 2 FC (y axis) of each gene following treatment with LPS ± RTA-404 in microglia in either Nfe2l2 fl/fl mice or Nfe2l2 ENDO mice were plotted. (iii): ∗P < 0.0001, F (1,2072) = 20.12 relates to main effect of RTA-404 on microglial gene sets suppressed by LPS, two-way ANOVA.

Article Snippet: Nrf2 flox mouse , The Jackson Laboratory , Cat. #25433.

Techniques: Activation Assay, Injection, Saline, Isolation, RNA Sequencing, Generated, Expressing, Modification, Gene Expression, Inhibition

Journal: iScience

Article Title: Endothelial cell Nrf2 controls neuroinflammation following a systemic insult

doi: 10.1016/j.isci.2025.112630

Figure Lengend Snippet: Endothelial cell Nrf2 is a key point of regulating astrogliosis under inflammatory conditions Mice were given i.p. injection of either saline or RTA-404 (n = 6–8) daily for 4 days, following which either saline or LPS was given for 24 h. The brain cells were isolated and then sorted with FACS for astrocytes, RNA extracted and RNA-seq performed. Scatterplots were generated for genes with average expression >0.1 FPKM across the datasets. Highlighted with red and blue crosses are the genes whose expression are significantly increased or decreased respectively ( DESeq2 P _adj < 0.05, n = 6–8). (A) Scatterplot of RNA-seq analysis showing the astrocyte transcriptome modified by peripheral LPS insult compared with saline in Nfe2l2 fl/fl mice (LPS vs Saline). (B) Scatterplot of RNA-seq analysis showing the LPS-induced astrocyte transcriptome modified by pre-administration of RTA-404 in Nfe2l2 fl/fl mice (LPS+RTA-404 vs. LPS+saline). (C) Scatterplot of RNA-seq analysis showing the LPS-induced astrocyte transcriptome modified by pre-administration of RTA-404 in Nfe2l2 ENDO mice. (D) IPA analysis identifying activated or inhibited pathways in astrocytes by pre-administration of RTA-404 under conditions of LPS exposure (LPS+RTA-404 vs. LPS+saline). Significantly DEGs ( DESeq2 P_adj < 0.05, ∣Log 2 FC∣>1, n = 6–8) with the astrocyte transcriptome as reference dataset were analyzed to calculate the p-value of overlap and Z score of overall activation/inhibition states of individual pathways. The significantly inhibited (p < 0.05, Z score <-2) pathways are shown in the bar chart in blue. (E) The influence of RTA-404 on LPS-induced genes in astrocytes. For LPS-induced genes (x axis, FPKM>1 , DESeqP_adj < 0.05, Log 2 FC > 1) in astrocytes in Nfe2l2 fl/fl mice, the Log 2 FC (y axis) of each gene following treatment with LPS ± RTA-404 in astrocytes in either Nfe2l2 fl/fl mice or Nfe2l2 ENDO mice were plotted. (iii) ∗p < 0.0001, F (1,6056) = 29.29 relates to main effect of RTA-404 on astrocytic gene sets induced by LPS, two-way ANOVA. (F) The influence of RTA-404 on LPS-repressed genes in astrocytes. For LPS-repressed genes (x axis, FPKM>1 , DESeqP _adj < 0.05, Log 2 FC < -1) in astrocytes in Nfe2l2 fl/fl mice, the Log 2 FC (y axis) of each gene following treatment with LPS ± RTA-404 in astrocytes in either Nfe2l2 fl/fl mice or Nfe2l2 ENDO mice were plotted. (iii): ∗p < 0.0001, F (1,25556) = 68.12 relates to main effect of RTA-404 on astrocytic gene sets suppressed by LPS, two-way ANOVA.

Article Snippet: Nrf2 flox mouse , The Jackson Laboratory , Cat. #25433.

Techniques: Injection, Saline, Isolation, RNA Sequencing, Generated, Expressing, Modification, Activation Assay, Inhibition

Journal: iScience

Article Title: Endothelial cell Nrf2 controls neuroinflammation following a systemic insult

doi: 10.1016/j.isci.2025.112630

Figure Lengend Snippet: Endothelial cell Nrf2 regulates immune cell brain infiltration under inflammatory conditions Mice were given i.p. injection of either saline or RTA-404 (n = 6–8) daily for 4 days, following which either saline or LPS was given for 24 h. The brain cells were analyzed using flow cytometer and the number of macrophages (CD11b + CD45 high ) in the brain following LPS or RTA-404 treatment were quantified. (A) Representative flow cytometry graphs showing microglia and macrophage population in the brain. (B) Statistical analysis showing the effect of either LPS or RTA-404 on peripheral monocyte brain infiltration. Data are represented as mean ± SE, p = 0.0023, 0.0010, 0.0045,0.4774 from left to right, two-way ANOVA plus Sidak post hoc (n = 6–8).

Article Snippet: Nrf2 flox mouse , The Jackson Laboratory , Cat. #25433.

Techniques: Injection, Saline, Flow Cytometry

Journal: BMC biotechnology

Article Title: Dexamethasone-boosted mesenchymal stem cell secretome: insight into hepatic protection.

doi: 10.1186/s12896-025-00980-8

Figure Lengend Snippet: Fig. 7 DEXA-treated BM-MSCs conditioned media decreased glucose uptake in hepatoma cells and reduced levels of hepatic inflammatory, angiogenic, and oxidative stress markers in mice. HepG2 cells treated with BM-MSC/DEXA-S significantly consumed less glucose compared with untreated control cells (P < 0.001) (A). Transfusion of BM-MSC-S in ALF mice variably reduced the inflammatory (TNF-α), and the angiogenic (VEGF) markers, and ameliorated the oxidative stress markers (Nrf2 and SOD) (C&D, respectively). Data are presented as mean (± SD)

Article Snippet: TNF-α and Nrf2 were determined by mouse TNF-α and Nrf2 ELISA kits (CUSABIO Cat. No. CSB-E04741m, and CSB-E16188m, respectively).

Techniques: Control