Journal: The FASEB Journal

Article Title: 4‐Octyl Itaconate Attenuates Age‐Induced Parotid Gland Dysfunction Through the Suppression of Fibrosis and Lipid Deposition by AMPKα/PPARα and the Inhibition of Oxidative Stress via the AMPKα/Nrf2 Signaling Pathway

doi: 10.1096/fj.202602956RR

Figure Lengend Snippet: 4‐OI alleviates function and structure impairments in the parotid gland of aged mice. (A) Salivary flow rates in different groups of mice ( n = 6). (B) Representative images and quantitative analysis of hematoxylin and eosin (H&E) staining in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (C) Representative images and quantitative analysis of Periodic acid–Schiff (PAS) staining in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (D) Representative images and quantitative analysis of senescence‐associated β‐galactosidase (SA‐β‐gal) staining in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm. (E, F) Representative Western blot and quantitative analysis the protein levels of P16 INK4A , P21, and P53 in the parotid gland of different groups of mice ( n = 3). (G) Representative images of immunofluorescent staining for F4/80 (red) and DAPI (blue) and quantitative analysis of F4/80 positive cells in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm. (H) Representative images of immunofluorescent staining for CD45 (red) and DAPI (blue) and quantitative analysis of CD45 positive cells in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm. (I, J) Representative images of immunofluorescent staining for KRT7 (red, a marker for luminal ductal cells), KRT5 (green, a marker for basal ductal cells), and DAPI (blue) and quantitative analysis of KRT7 positive cells and KRT5 positive cells in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (K, L) Representative images of immunofluorescent staining for KRT14 (red, a marker for myoepithelial cells), BHLHA15 (green, a marker for mature acinar cells), and DAPI (blue) and quantitative analysis of KRT14 positive cells and BHLHA15 positive cells in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (M, N) Representative Western blot and quantitative analysis the protein levels of IRG1 in the parotid gland of different groups of mice ( n = 3). (O) Representative images of immunofluorescent staining for IRG1 (red), BHLHA15 (green), and DAPI (blue) in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). The data are presented as the means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. ns, not significant.

Article Snippet: The mice in the 13 M + 4‐OI group were administered 4‐OI (25 mg/kg/day, HY‐112675, MedChemExpress, NJ, USA) via intraperitoneal (i.p.) injection for 4 weeks, whereas the mice in the 13 M group and 2 M groups were given vehicle by i.p. injection.

Techniques: Analysis, Staining, Western Blot, Marker

Journal: The FASEB Journal

Article Title: 4‐Octyl Itaconate Attenuates Age‐Induced Parotid Gland Dysfunction Through the Suppression of Fibrosis and Lipid Deposition by AMPKα/PPARα and the Inhibition of Oxidative Stress via the AMPKα/Nrf2 Signaling Pathway

doi: 10.1096/fj.202602956RR

Figure Lengend Snippet: 4‐OI decreased age‐induced apoptosis, inflammation, and NLRP3 activation in the parotid gland of aged mice. (A, B) Representative Western blot and quantitative analysis the protein levels of IL‐1β and IL‐6 in the parotid gland of different groups of mice ( n = 3). (C) Representative images of immunofluorescent staining for TUNEL (red) and DAPI (blue) and quantitative analysis of TUNEL positive cells in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm. (D) Flow cytometry to detect cell apoptosis. (E, F) Representative Western blot and quantitative analysis the protein levels of Bcl‐2, Bax, Cleaved‐Caspase3 in the parotid gland of different groups of mice ( n = 3). (G, H) Representative Western blot and quantitative analysis the protein levels of NLRP3, ASC, and Caspase 1/p20 in the parotid gland of different groups of mice ( n = 3). (I) Representative images of immunofluorescent staining for NLRP3 (red), BHLHA15 (green), and DAPI (blue) in the parotid gland of different groups of mice. Scale bars: 50 μm (top) and 20 μm (bottom). The white line with arrows indicates the region selected for colocalization analysis. (J) Fluorescence intensity profiles of NLRP3 (red) and BHLHA15 (green) along the white line shown in (I). The x‐axis represents the distance (μm) along the white line with arrows, and the y‐axis represents fluorescence intensity (a.u.). A shorter distance between the peak intensities of the two channels indicates a higher degree of colocalization. The data are presented as the means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. ns, not significant.

Article Snippet: The mice in the 13 M + 4‐OI group were administered 4‐OI (25 mg/kg/day, HY‐112675, MedChemExpress, NJ, USA) via intraperitoneal (i.p.) injection for 4 weeks, whereas the mice in the 13 M group and 2 M groups were given vehicle by i.p. injection.

Techniques: Activation Assay, Western Blot, Analysis, Staining, TUNEL Assay, Flow Cytometry, Fluorescence

Journal: The FASEB Journal

Article Title: 4‐Octyl Itaconate Attenuates Age‐Induced Parotid Gland Dysfunction Through the Suppression of Fibrosis and Lipid Deposition by AMPKα/PPARα and the Inhibition of Oxidative Stress via the AMPKα/Nrf2 Signaling Pathway

doi: 10.1096/fj.202602956RR

Figure Lengend Snippet: 4‐OI inhibits fibrosis, excessive production of ROS, and reversed aged‐induced mitochondrial membrane potential decreased in the parotid gland of aged mice. (A) Representative images and quantitative analysis of Masson staining in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (B) Representative images and quantitative analysis of Sirius Red staining in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (C, D) Representative Western blot and quantitative analysis the protein levels of FN1 and COL1 in the parotid gland of different groups of mice ( n = 3). (E) Representative images of immunofluorescent staining for COL1 (red) and DAPI (blue) and quantitative analysis of relative fluorescence intensity of COL1 in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm. (F) Representative Western blot and quantitative analysis the protein levels of αSMA in the parotid gland of different groups of mice ( n = 3). (G) Representative ROS‐induced fluorescence staining images labeled with DHE dye (red) and DAPI (blue), accompanied by quantitative analysis of relative DHE fluorescence intensity in the parotid glands of mice from different groups ( n = 6). (H) Representative images and quantitative analysis of JC‐1 staining in the parotid glands of mice from different groups ( n = 6). Scale bar: 50 μm (top) and 20 μm (bottom). (I, J) Representative Western blot and quantitative analysis the protein levels of PGC‐1α, TFAM, and cytochrome c in the parotid gland of different groups of mice ( n = 3). The data are presented as the means ± SD. ** p < 0.01, *** p < 0.001. DHE, dihydroethidium; ns, not significant; ROS, reactive oxygen species.

Article Snippet: The mice in the 13 M + 4‐OI group were administered 4‐OI (25 mg/kg/day, HY‐112675, MedChemExpress, NJ, USA) via intraperitoneal (i.p.) injection for 4 weeks, whereas the mice in the 13 M group and 2 M groups were given vehicle by i.p. injection.

Techniques: Membrane, Analysis, Staining, Western Blot, Fluorescence, Labeling

Journal: The FASEB Journal

Article Title: 4‐Octyl Itaconate Attenuates Age‐Induced Parotid Gland Dysfunction Through the Suppression of Fibrosis and Lipid Deposition by AMPKα/PPARα and the Inhibition of Oxidative Stress via the AMPKα/Nrf2 Signaling Pathway

doi: 10.1096/fj.202602956RR

Figure Lengend Snippet: Effect of 4‐OI on the metabolomics and transcriptome profile of aged mice parotid glands and enhancement of AMPKα and Nrf2 pathway in the parotid gland of aged mice by 4‐OI. OPLS‐DA score plot of metabolites detected in (A) 13 M versus 2 M and (B) 13 M + 4‐OI versus 13 M. (C) The Venn diagram illustrates the differentially expressed metabolites that are particular or common within different comparison groups. (D) KEGG pathway classifications on differential metabolites. The vertical axis represents KEGG compound categories, while the horizontal axis shows the number of compounds annotated to each category. Bar colors indicate the primary classification level of the compounds. Volcano plots plot of differential metabolites detected in (E) 13 M versus 2 M and (F) 13 M + 4‐OI versus 13 M. (G) Histogram displaying the numbers of differential metabolites in 13 M versus 2 M and 13 M + 4‐OI versus 13 M. Heatmap showing the variable importance (VIP) scores of the top 30 differential metabolites in (H) 13 M versus 2 M and (I) 13 M + 4‐OI versus 13 M. Bubble plots illustrate the primary metabolic pathways responsible for the enrichment of differential metabolites in (J) 13 M versus 2 M and (K) 13 M + 4‐OI versus 13 M. Volcano plots plot of DEGs detected in (L) 13 M versus 2 M and (M) 13 M + 4‐OI versus 13 M. (N) Histogram of the numbers of up‐regulated and downregulated DEGs in 13 M versus 2 M and 13 M + 4‐OI versus 13 M. Bubble plots illustrate the primary KEGG pathways responsible for the enrichment of DEGs in (O) 13 M versus 2 M and (P) 13 M + 4‐OI versus 13 M. (Q, R) Representative Western blot and quantitative analysis the protein levels of AMPKα, p‐AMPKα, and PPARα in the parotid gland of different groups of mice ( n = 3). (S, T) Representative Western blot and quantitative analysis the protein levels of Nrf2, Keap1, NQO‐1, and HO‐1 in the parotid gland of different groups of mice ( n = 3). The data are presented as the means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. ns, not significant.

Article Snippet: The mice in the 13 M + 4‐OI group were administered 4‐OI (25 mg/kg/day, HY‐112675, MedChemExpress, NJ, USA) via intraperitoneal (i.p.) injection for 4 weeks, whereas the mice in the 13 M group and 2 M groups were given vehicle by i.p. injection.

Techniques: Comparison, Western Blot, Analysis

Journal: The FASEB Journal

Article Title: 4‐Octyl Itaconate Attenuates Age‐Induced Parotid Gland Dysfunction Through the Suppression of Fibrosis and Lipid Deposition by AMPKα/PPARα and the Inhibition of Oxidative Stress via the AMPKα/Nrf2 Signaling Pathway

doi: 10.1096/fj.202602956RR

Figure Lengend Snippet: The binding mode of 4‐OI with AMPKα through molecular docking and molecular dynamics simulations. (A) A three‐dimensional model of the complex. (B) the specific binding mode between 4‐OI and AMPKα. (C) The two‐dimensional structure of the binding mode during the kinetic simulation of 4‐OI and AMPKα. (D) The root mean square deviation (RMSD) of 4‐OI (blue) with AMPKα (black). (E) The root mean square fluctuation (RMSF) of 4‐OI with AMPKα. (F) Binding free energy components are detailed, including ΔVDWAALS, ΔEEL, ΔEPB, ΔENPOLAR, ΔGGAS, ΔGSOLV, ΔTOTAL. (G) The per‐residue decomposition energy plot of critical amino acid residues. (H–J) The radius of gyration (Rg), solvent accessible surface area (SASA), and hydrogen bond number of 4‐OI with AMPKα. (K, L) Two‐ and three‐dimensional free energy landscape (FEL) plots quantitatively depicting conformational energy states. Energy troughs (dark blue regions) correspond to high‐probability conformational clusters and represent the system's stable states. Energy plateaus or saddle points (yellow/red regions) indicate transition states or energy barriers that separate different conformational clusters, reflecting the kinetic bottlenecks of conformational transitions.

Article Snippet: The mice in the 13 M + 4‐OI group were administered 4‐OI (25 mg/kg/day, HY‐112675, MedChemExpress, NJ, USA) via intraperitoneal (i.p.) injection for 4 weeks, whereas the mice in the 13 M group and 2 M groups were given vehicle by i.p. injection.

Techniques: Binding Assay, Docking Assay, Residue, Solvent

Journal: The FASEB Journal

Article Title: 4‐Octyl Itaconate Attenuates Age‐Induced Parotid Gland Dysfunction Through the Suppression of Fibrosis and Lipid Deposition by AMPKα/PPARα and the Inhibition of Oxidative Stress via the AMPKα/Nrf2 Signaling Pathway

doi: 10.1096/fj.202602956RR

Figure Lengend Snippet: PPARα antagonist verified that 4‐OI alleviates parotid gland lipids accumulation via the PPARα‐mediated FAO signaling pathway in aged mice. (A) Representative images and quantitative analysis of hematoxylin and eosin (H&E) staining in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (B) Representative images and quantitative analysis of Oil Red O staining and BODIPY staining in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm. (C) Representative IHC staining images and quantitative analysis of the ACOX1 and CPT1A in the parotid gland of different groups of mice. Scale bar: 50 μm. (D, E) Representative Western blot and quantitative analysis the protein levels of ACOX1 and CPT1A in the parotid gland of different groups of mice ( n = 3). (F) Representative images of immunofluorescent staining for CPT1A (red), ACOX1 (green), PPARα (pink), and DAPI (blue) in the parotid gland of different groups of mice. Scale bars: 50 μm (top) and 20 μm (bottom). The white line with arrows indicates the region selected for colocalization analysis. (G) Fluorescence intensity profiles of CPT1A (red), ACOX1 (green), and PPARα (pink) along the white line shown in (F). The x‐axis represents the distance (μm) along the white line with arrows, and the y‐axis represents fluorescence intensity (a.u.). A shorter distance between the peak intensities of the three channels indicates a higher degree of colocalization. (H) Representative images of immunofluorescent staining for COL1 (red) and DAPI (blue) and quantitative analysis of relative fluorescence intensity of COL1 in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm. (I, J) Representative Western blot and quantitative analysis the protein levels of ACC, p‐ACC, and PPARα in the parotid gland of different groups of mice ( n = 3). The data are presented as the means ± SD.* p < 0.05 ** p < 0.01, *** p < 0.001. ns, not significant.

Article Snippet: The mice in the 13 M + 4‐OI group were administered 4‐OI (25 mg/kg/day, HY‐112675, MedChemExpress, NJ, USA) via intraperitoneal (i.p.) injection for 4 weeks, whereas the mice in the 13 M group and 2 M groups were given vehicle by i.p. injection.

Techniques: Analysis, Staining, Immunohistochemistry, Western Blot, Fluorescence

Journal: The FASEB Journal

Article Title: 4‐Octyl Itaconate Attenuates Age‐Induced Parotid Gland Dysfunction Through the Suppression of Fibrosis and Lipid Deposition by AMPKα/PPARα and the Inhibition of Oxidative Stress via the AMPKα/Nrf2 Signaling Pathway

doi: 10.1096/fj.202602956RR

Figure Lengend Snippet: Nrf2 inhibitor reverses 4‐OI's restoration of function and structure impairments in parotid gland in aged mice. (A) Representative images and quantitative analysis of hematoxylin and eosin (H&E) staining in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (B) Salivary flow rates in different groups of mice ( n = 6). (C, D) Representative Western blot and quantitative analysis the protein levels of Nrf2, Keap1, NQO‐1, and HO‐1 in the parotid gland of different groups of mice ( n = 3). (E) Representative images of immunofluorescent staining for Nrf2 (red) and DAPI (blue) and quantitative analysis of relative fluorescence intensity of Nrf2 in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm. The data are presented as the means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. ns, not significant.

Article Snippet: The mice in the 13 M + 4‐OI group were administered 4‐OI (25 mg/kg/day, HY‐112675, MedChemExpress, NJ, USA) via intraperitoneal (i.p.) injection for 4 weeks, whereas the mice in the 13 M group and 2 M groups were given vehicle by i.p. injection.

Techniques: Analysis, Staining, Western Blot, Fluorescence

Journal: The FASEB Journal

Article Title: 4‐Octyl Itaconate Attenuates Age‐Induced Parotid Gland Dysfunction Through the Suppression of Fibrosis and Lipid Deposition by AMPKα/PPARα and the Inhibition of Oxidative Stress via the AMPKα/Nrf2 Signaling Pathway

doi: 10.1096/fj.202602956RR

Figure Lengend Snippet: AMPKα inhibitor reverses 4‐OI's reduction of fibrosis in parotid gland in aged mice. (A) Representative images and quantitative analysis of hematoxylin and eosin (H&E) staining in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (B) Representative images of immunofluorescent staining for KRT7 (red, a marker for luminal ductal cells), KRT5 (green, a marker for basal ductal cells), and DAPI (blue) and (D) quantitative analysis of KRT7 positive cells and KRT5 positive cells in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (C) Representative images of immunofluorescent staining for KRT14 (red, a marker for myoepithelial cells), BHLHA15 (green, a marker for mature acinar cells), and DAPI (blue) and (E) quantitative analysis of KRT14 positive cells and BHLHA15 positive cells in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (F) Representative images and (I) quantitative analysis of Masson staining in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (G) Representative images and (J) quantitative analysis of Sirius Red staining in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm (top) and 20 μm (bottom). (H) Representative images of immunofluorescent staining for COL1 (red) and DAPI (blue) and (K) quantitative analysis of relative fluorescence intensity of COL1 in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm. (L–M) Representative Western blot and quantitative analysis the protein levels of FN1, COL1, and TGF‐β in the parotid gland of different groups of mice ( n = 3). (N) Representative Western blot and quantitative analysis the protein levels of αSMA in the parotid gland of different groups of mice ( n = 3). (O) Representative images of immunofluorescent staining for αSMA (red), VIMENTIN (green), and DAPI (blue) in the parotid gland of different groups of mice. Scale bars: 50 μm (top) and 20 μm (bottom). The white line with arrows indicates the region selected for colocalization analysis. (P) Fluorescence intensity profiles of αSMA (red) and VIMENTIN (green) along the white line shown in (O). The x ‐axis represents the distance (μm) along the white line with arrows, and the y ‐axis represents fluorescence intensity (a.u.). A shorter distance between the peak intensities of the two channels indicates a higher degree of colocalization. The data are presented as the means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. ns, not significant.

Article Snippet: The mice in the 13 M + 4‐OI group were administered 4‐OI (25 mg/kg/day, HY‐112675, MedChemExpress, NJ, USA) via intraperitoneal (i.p.) injection for 4 weeks, whereas the mice in the 13 M group and 2 M groups were given vehicle by i.p. injection.

Techniques: Analysis, Staining, Marker, Fluorescence, Western Blot

Journal: The FASEB Journal

Article Title: 4‐Octyl Itaconate Attenuates Age‐Induced Parotid Gland Dysfunction Through the Suppression of Fibrosis and Lipid Deposition by AMPKα/PPARα and the Inhibition of Oxidative Stress via the AMPKα/Nrf2 Signaling Pathway

doi: 10.1096/fj.202602956RR

Figure Lengend Snippet: AMPKα inhibitor reverses 4‐OI's alleviation of apoptosis, inflammation, and mitochondrial dysfunction in parotid gland in aged mice. (A) Representative ROS‐induced fluorescence staining images labeled with DHE dye (red) and DAPI (blue), accompanied by (C) quantitative analysis of relative DHE fluorescence intensity in the parotid glands of mice from different groups ( n = 6). (B) Representative images and (D) quantitative analysis of JC‐1 staining in the parotid glands of mice from different groups ( n = 6). Scale bar: 50 μm (top) and 20 μm (bottom). (E) Representative transmission electron microscopy image of mitochondria in the parotid glands of 2 M, 13 M, 13 M + 4‐OI, and 13 M + 4‐OI + CC groups. Scale bars: 1 μm (top) and 500 nm (bottom). (F, G) Representative Western blot and quantitative analysis the protein levels of PGC‐1α, TFAM, and cytochrome c in the parotid gland of different groups of mice ( n = 3). (H, I) Representative Western blot and quantitative analysis the protein levels of AMPKα, p‐AMPKα, SIRT1, GSK3β, p‐GSK3β, Nrf2, and Keap1 in the parotid gland of different groups of mice ( n = 3). (J) Representative images of immunofluorescent staining for Nrf2 (red) and DAPI (blue) and HO‐1 (red) and DAPI (blue) in the parotid gland of different groups of mice. Scale bars: 50 μm. (K, L) Representative Western blot and quantitative analysis the protein levels of TLR4, NF‐κB p65, p‐NF‐κB p65, IκB‐α, and p‐IκB‐α in the parotid gland of different groups of mice ( n = 3). (M, N) Representative Western blot and quantitative analysis the protein levels of NLRP3, TNF‐α, P16 INK4A , P21, and P53 in the parotid gland of different groups of mice ( n = 3). (O) Representative images and quantitative analysis of senescence‐associated β‐galactosidase (SA‐β‐gal) staining in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm. (P) Representative images of immunofluorescent staining for 8‐OHdG (red) and DAPI (blue) in the parotid gland of different groups of mice. Scale bars: 50 μm. (Q) Representative images of immunofluorescent staining for TUNEL (red) and DAPI (blue) and quantitative analysis of TUNEL positive cells in the parotid gland of different groups of mice ( n = 6). Scale bars: 50 μm. The data are presented as the means ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. DHE, dihydroethidium; ns, not significant; ROS, reactive oxygen species.

Article Snippet: The mice in the 13 M + 4‐OI group were administered 4‐OI (25 mg/kg/day, HY‐112675, MedChemExpress, NJ, USA) via intraperitoneal (i.p.) injection for 4 weeks, whereas the mice in the 13 M group and 2 M groups were given vehicle by i.p. injection.

Techniques: Fluorescence, Staining, Labeling, Analysis, Transmission Assay, Electron Microscopy, Western Blot, TUNEL Assay

Journal: International Journal of Nanomedicine

Article Title: Synergistic Neuroprotection of Fe 3 O 4 Nanoparticles Combined with Intermittent Theta Burst Stimulation in Ischemic Stroke via Nrf2-Mediated Ferroptosis Regulation

doi: 10.2147/IJN.S574711

Figure Lengend Snippet: Therapeutic mechanism of Fe 3 O 4 for precise ischemic stroke management.

Article Snippet: In vivo,The 4-8-week-old healthy male Sprague-Dawley (SD) rats were randomly divided into eight groups: 1) Sham group, 2) Saline group, 3) MCAO/R group, 4) iTBS group, 5) rTMS group, 6) Fe 3 O 4 +iTBS group, 7) Fe 3 O 4 +rTMS group, 8) Fe 3 O 4 +magnet group.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Synergistic Neuroprotection of Fe 3 O 4 Nanoparticles Combined with Intermittent Theta Burst Stimulation in Ischemic Stroke via Nrf2-Mediated Ferroptosis Regulation

doi: 10.2147/IJN.S574711

Figure Lengend Snippet: Fabrication and characterization of Fe 3 O 4 . ( A ) Representative TEM images of prepared NPs. ( B ) zeta potential measurements (n = 3). ( C ) Size distributions of Fe 3 O 4 by dynamic light scattering (n = 3). ( D ) Confirmation of magnetic targeting of Fe 3 O 4 by static magnetic field. Red circles indicate nanoparticle aggregation toward the magnetic field. ( E ) Fe 3 O 4 labeled with iR780 by fluorescence images. ( F ) Characterization of Fe 3 O 4 NPs using EDS.

Article Snippet: In vivo,The 4-8-week-old healthy male Sprague-Dawley (SD) rats were randomly divided into eight groups: 1) Sham group, 2) Saline group, 3) MCAO/R group, 4) iTBS group, 5) rTMS group, 6) Fe 3 O 4 +iTBS group, 7) Fe 3 O 4 +rTMS group, 8) Fe 3 O 4 +magnet group.

Techniques: Zeta Potential Analyzer, Labeling, Fluorescence

Journal: International Journal of Nanomedicine

Article Title: Synergistic Neuroprotection of Fe 3 O 4 Nanoparticles Combined with Intermittent Theta Burst Stimulation in Ischemic Stroke via Nrf2-Mediated Ferroptosis Regulation

doi: 10.2147/IJN.S574711

Figure Lengend Snippet: Study on BBB penetrability, biodistribution and biosafety of Fe 3 O 4 NPs. ( A ) Hemolysis test of Fe 3 O 4 NPs in different concentrations. ( B ) Fluorescence images of the heads of MCAO/R rats after intravenous (iv.) injection with iR780-labeled NPs at 6 h. ( C ) Fluorescence images of brain sections at 0, 24, 48, 72, 96 and 120 h post-injection. Fluorescence signals were quantified using Living Image software with background subtraction. Data represent transient tight junction remodeling and enhanced permeability, and no statistical analysis was performed. ( D ) Radiant efficiency was quantified at 0, 24, 48, 72, 96, and 120 h post-injection. Data represent a single subject. ( E ) Fluorescence images of the dissected main organs and brains at 48 h post-injection. ( F ) Representative H&E staining of ischemic brain sections after different treatments. Scale bar = 20 μM. ( G ) Prussian blue staining of tumors after iv. injection 3 days of Fe 3 O 4 NPs. Scale bar: 20 μM and ( H ) their quantitative analyses (n = 5). Data are presented as means ± SD (n = 5). Statistical significance was determined by one-way ANOVA with a Tukey post hoc test. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: In vivo,The 4-8-week-old healthy male Sprague-Dawley (SD) rats were randomly divided into eight groups: 1) Sham group, 2) Saline group, 3) MCAO/R group, 4) iTBS group, 5) rTMS group, 6) Fe 3 O 4 +iTBS group, 7) Fe 3 O 4 +rTMS group, 8) Fe 3 O 4 +magnet group.

Techniques: Fluorescence, IV Injection, Labeling, Injection, Software, Permeability, Staining

Journal: International Journal of Nanomedicine

Article Title: Synergistic Neuroprotection of Fe 3 O 4 Nanoparticles Combined with Intermittent Theta Burst Stimulation in Ischemic Stroke via Nrf2-Mediated Ferroptosis Regulation

doi: 10.2147/IJN.S574711

Figure Lengend Snippet: Neuroprotective effects and modulation of microglia phenotypes in vitro. ( A ) and ( B ) The cell viability of SH-SY5Y cells and BV2 cells after OGD/R or incubation with Fe 3 O 4 of different concentration for 12 h (n=2). ( C ) Fluorescence images of ROS in different groups after OGD/R (n = 3). Scale bar: 40 μM. ( D ) Immunofluorescence co-staining images of BV2 cells showing M1 phenotype marker CD86 (green) (n = 3). Scale bar: 20 μM, corresponding quantitative fluorescence of ROS ( E ) and CD86 (F). ( G ) The changes of Δ ψ m in SH-SY5Y cells undergoing OGD/R after different treatments for 48 h and ( H ) their quantitative analyses (n = 3). Relative expression of pro-inflammatory IL-6 ( I ) and TNF-α (J), and MDA ( K ) in DMEN from BV2 cell culture. Data are presented as means ± SD (n = 3). ***P < 0.001, **P < 0.01, *P < 0.05.

Article Snippet: In vivo,The 4-8-week-old healthy male Sprague-Dawley (SD) rats were randomly divided into eight groups: 1) Sham group, 2) Saline group, 3) MCAO/R group, 4) iTBS group, 5) rTMS group, 6) Fe 3 O 4 +iTBS group, 7) Fe 3 O 4 +rTMS group, 8) Fe 3 O 4 +magnet group.

Techniques: In Vitro, Incubation, Concentration Assay, Fluorescence, Immunofluorescence, Staining, Marker, Expressing, Cell Culture