Journal: Molecular Medicine Reports

Article Title: Cigarette smoke combined with Klebsiella pneumoniae induce damage to the air-blood barrier in chronic obstructive pulmonary disease rats via the MAPK/NF-κB/IκBα pathway

doi: 10.3892/mmr.2026.13797

Figure Lengend Snippet: Injury of mucus hypersecretion and oxidative damage in all groups. (A) AB-PAS staining images of the airway in all groups (AB-PAS; ×200 magnification and ×400 magnification). Arrows indicate goblet cells. (B) The expression level of MUC5AC and MUC5B in the airway tested using immunohistochemistry (×200 magnification). (C) The expression of IL-6, IL-10, TNF-α, T-SOD, MDA, MMP-9 and TIMP-1 in lung tissue and serum of rats in all groups. The values are expressed as mean ± SE; (n=8). *P<0.05 and **P<0.01 vs. normal group. AB-PAS, Alcian blue-periodic acid-schiff; MUC5AC, mucoprotein-5AC; MUC5B, mucoprotein-5B; T-SOD, total superoxide dismutase; MDA, malondialdehyde; MMP-9, matrix metalloproteinase 9; TIMP-1, tissue inhibitor of metalloproteinases 2; IOD, integrated optical density.

Article Snippet: The secretion of IL-6 (Rat IL-6 ELISA Kit; cat. no. E-EL-R0015; Elabscience Bionovation Inc.), IL-10 (Rat IL-10 ELISA Kit; cat. no. E-EL-R0016; Elabscience Bionovation Inc.), malondialdehyde (MDA) (MDA assay kit; cat. no. A003-1-2; Nanjing Jiancheng Bioengineering Institute), total superoxide dismutase (T-SOD) (SOD assay kit; cat. no. A001-3-2; Nanjing Jiancheng Bioengineering Institute), MMP-9 (Rat MMP-9 ELISA Kit; cat. no. E-EL-R3021; Elabscience Bionovation Inc.) and tissue Inhibitors of metalloproteinase 1 (TIMP-1 ELISA Kit; cat. no. E-EL-R0540; Elabscience Bionovation Inc.) in the lung tissue homogenate was measured using ELISA kits, following the manufacturer's instructions.

Techniques: Staining, Expressing, Immunohistochemistry

Journal: Signal Transduction and Targeted Therapy

Article Title: Human adipose and umbilical cord mesenchymal stem cell-derived extracellular vesicles mitigate photoaging via TIMP1/Notch1

doi: 10.1038/s41392-024-01993-z

Figure Lengend Snippet: AMSC-EV and HUMSC-EV mitigated photoaging of HDFs in vitro. a Representative images of MSC-EV uptake by HDFs pretreated with or without UVB irradiation (scale bar, 20 μm). b Representative immunofluorescence staining images ROS (green) and DAPI (scale bar, 20 μm). c Fluorescence intensity of ROS levels. n = 3, *** P < 0.001, **** P < 0.0001. d Representative images of SA-β-gal staining in HDFs (scale bar, 100 μm). e Quantitation of SA-β-gal positive cells in HDFs. n = 3, **** P < 0.0001. f Representative immunofluorescence staining images of positive cells of γ-H2Ax (red) and DAPI (scale bar, 20 μm). g Quantitation of a mean number of γ-H2Ax foci/cell. h Quantitation of HDFs proliferation detected by CCK8 assay with the OD value on Day 1, Day 2, Day 3, Day 4, and Day 5. n = 3, *** P < 0.001, **** P < 0.0001. i Quantitative of COL31A in HDFs by qRT-PCR. n = 3, * P < 0.05, ** P < 0.01. j Quantitation of TIMP1, MMP1, and MMP9 released by HDFs by qRT-PCR. n = 3, * P < 0.05, *** P < 0.001, **** P < 0.0001. k Western blot analysis showing the change of COL3, TIMP1, MMP1 and MMP9 in HDFs. l Quantitative of COL3, TIMP1, MMP1, and MMP9 in HDFs by western blot. n = 3, * P < 0.05, ** P < 0.01, *** P < 0.001. m Quantitation of IL-1β, IL-6, and TNF-α released by HDFs by qRT-PCR. n = 3, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. n Quantitation of IL-1β, IL-6, and TNF-α released by HDFs was detected through ELISA. n = 3, ** P < 0.01, **** P < 0.0001

Article Snippet: After UVB, HDFs were incubated with ADAM10 inhibitor GI254023X (HY-19956, MedChemExpress) for 2 h, and then cells were incubated with fibroblast medium for an additional 24 h. After UVB, 2 mM Notch activator Valproic acid (Sodium Valproate) sodium (HY-10585A, MedChemExpress) were used with MSC-EV or TIMP1 for 24 h.

Techniques: In Vitro, Irradiation, Immunofluorescence, Staining, Fluorescence, Quantitation Assay, CCK-8 Assay, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: Human adipose and umbilical cord mesenchymal stem cell-derived extracellular vesicles mitigate photoaging via TIMP1/Notch1

doi: 10.1038/s41392-024-01993-z

Figure Lengend Snippet: Histological analysis and inflammation antibody array of the dorsal skin in nude mice after UVB irradiation and EV treatment. a Representative images of Masson’s staining (scale bar, 50 μm). b Representative images of picrosirius red staining (scale bar, 50 μm). c Quantitation of collage area from Masson’s staining statistics. n = 6, ** P < 0.01, *** P < 0.001. d Quantitation of collage area from picrosirius red staining statistics. n = 6, *** P < 0.001. e Western blot analysis for COL1, COL3, TIMP1, MMP1 and MMP3. f The top inflammatory factors examined by Inflammation Antibody Array. g Heatmap highlighting the representative inflammatory factors examined by the Inflammation Antibody Array. h Western blot analysis for inflammatory factors IL-1β, IL-6, TNF-α, and GM-CSF

Article Snippet: After UVB, HDFs were incubated with ADAM10 inhibitor GI254023X (HY-19956, MedChemExpress) for 2 h, and then cells were incubated with fibroblast medium for an additional 24 h. After UVB, 2 mM Notch activator Valproic acid (Sodium Valproate) sodium (HY-10585A, MedChemExpress) were used with MSC-EV or TIMP1 for 24 h.

Techniques: Ab Array, Irradiation, Staining, Quantitation Assay, Western Blot

Journal: Signal Transduction and Targeted Therapy

Article Title: Human adipose and umbilical cord mesenchymal stem cell-derived extracellular vesicles mitigate photoaging via TIMP1/Notch1

doi: 10.1038/s41392-024-01993-z

Figure Lengend Snippet: MSC-EV rescues HDFs and HaCaTs photoaging by upregulating TIMP1. a Venn diagram of the protein contents of AMSC-EV and HUMSC-EV. b Heatmap of the protein contents of AMSC-EV and HUMSC-EV. c Heatmap of extracellular matrix organization. d Western blot analysis showing COL1, COL3, COL6, and TIMP1 expression in AMSC-EV and HUMSC-EV. e Quantitation of TIMP1 in AMSC-EV and HUMSC-EV was detected by Western blot. n = 3. f Representative immunofluorescence staining images of positive cells of ROS (green) and DAPI (scale bar, 35 μm). g Quantitation of ROS-positive cells. n = 3, **** P < 0.0001. h Representative immunofluorescence staining images of positive cells of γ-H2Ax (red) and DAPI (scale bar, 20 μm). i Quantitation of a mean number of γ-H2Ax foci/cell. n = 3, *** P < 0.001, **** P < 0.0001. j Representative images of SA-β-gal staining in HDFs and HaCaTs (scale bar, 100 μm). k Quantitation of SA-β-gal positive cells in HDFs and HaCaTs. n = 3, *** P < 0.001, **** P < 0.0001. l Representative images of transwell assays of HaCaTs (scale bar, 100 μm). m epresentative images of migration assay of HaCaTs (scale bar, 100 μm). n Western blot analysis showing the change of COL3, TIMP1, MMP1 and MMP9 in HDFs

Article Snippet: After UVB, HDFs were incubated with ADAM10 inhibitor GI254023X (HY-19956, MedChemExpress) for 2 h, and then cells were incubated with fibroblast medium for an additional 24 h. After UVB, 2 mM Notch activator Valproic acid (Sodium Valproate) sodium (HY-10585A, MedChemExpress) were used with MSC-EV or TIMP1 for 24 h.

Techniques: Western Blot, Expressing, Quantitation Assay, Immunofluorescence, Staining, Migration

Journal: Signal Transduction and Targeted Therapy

Article Title: Human adipose and umbilical cord mesenchymal stem cell-derived extracellular vesicles mitigate photoaging via TIMP1/Notch1

doi: 10.1038/s41392-024-01993-z

Figure Lengend Snippet: MSC-EV rescues HDFs photoaging by downregulating NOTCH signaling pathway. a Venn diagram showing the overlapped genes between photoaging (UVB/Control) and Treatment (EV/UVB) based on data from RNA-seq. Rev-photoaging-DEGs were defined as subsets of overlapped DEGs that were changed in the opposite direction in photoaging and treatment. b Representative GO terms of Rev-photoaging DEGs for Biological Process (green), Cellular Component (orange), and Molecular Function (blue) were shown. c Heatmaps showing the expression profile of genes in response to UVB and EV treatment in Notch signaling. d Quantitation of different mRNA levels of Hes1, Tle1, Lfng, Dll1, and HeyL. e Western blot analysis showing expression of NOTCH1, NICD1, HES1, SIRT1, P16, P21, P53 and GAPDH after EV treatment. f Western blot analysis showing expression of NOTCH1, NICD1, HES1, SIRT1, P16, P21, P53 and GAPDH after TIMP1 treatment. g Representative immunofluorescence staining images of positive cells of HES1 in nude mice dorsal skin injected PBS, AMSC-EV, or HUMSC-EV (top, scale bar, 30 μm; bottom, scale bar, 10 μm). h Representative immunofluorescence staining images of SA-β-gal (scale bar, 100 μm), ROS (green), and γ-H2Ax (red) (scale bar, 30 μm). i Luciferase-reporter assays of HES1 transcriptional activity in HDFs after UVB, FM (culture medium), EV, TIMP1, GI254023X, and VPA treatment. n = 3, ** P < 0.01, *** P < 0.001, **** P < 0.0001. j Schematic of the role of EV in mediating UVB-induced photoaging of Notch signaling and cellular senescence. (Created with BioRender.com)

Article Snippet: After UVB, HDFs were incubated with ADAM10 inhibitor GI254023X (HY-19956, MedChemExpress) for 2 h, and then cells were incubated with fibroblast medium for an additional 24 h. After UVB, 2 mM Notch activator Valproic acid (Sodium Valproate) sodium (HY-10585A, MedChemExpress) were used with MSC-EV or TIMP1 for 24 h.

Techniques: Control, RNA Sequencing, Expressing, Quantitation Assay, Western Blot, Immunofluorescence, Staining, Injection, Luciferase, Activity Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: Human adipose and umbilical cord mesenchymal stem cell-derived extracellular vesicles mitigate photoaging via TIMP1/Notch1

doi: 10.1038/s41392-024-01993-z

Figure Lengend Snippet: Schematic illustrations of AMSC-EV and HUMSC-EV repairments on photoaging in vitro and in vivo via regulation of the TIMP1/Notch pathway (Created with BioRender.com). AMSC-EV and HUMSC-EV improve photoaging in UVB-irradiated nude mice, HaCaTs, HKCs, HDFs, and T-Skin models through anti-inflammatory effects, ECM remodeling, and anti-senescence properties. Mechanistically, proteomic analysis revealed that TIMP1 is highly expressed in both AMSC-EV and HUMSC-EV and exerts similar effects to MSC-EV. TIMP1 inhibits MMPs, thereby altering extracellular matrix remodeling. Additionally, TIMP1 downregulates the activated Notch pathway and inhibits its downstream targets Hes1, P16, P21, and P53

Article Snippet: After UVB, HDFs were incubated with ADAM10 inhibitor GI254023X (HY-19956, MedChemExpress) for 2 h, and then cells were incubated with fibroblast medium for an additional 24 h. After UVB, 2 mM Notch activator Valproic acid (Sodium Valproate) sodium (HY-10585A, MedChemExpress) were used with MSC-EV or TIMP1 for 24 h.

Techniques: In Vitro, In Vivo, Irradiation