Journal: Frontiers in Pharmacology

Article Title: Sodium nitrate regulates senescence accompanied by aortic atherosclerosis in ApoE −/− mice through the miR-34a/FGF-21 axis

doi: 10.3389/fphar.2025.1562321

Figure Lengend Snippet: Effect of sodium nitrate on H 2 O 2 -induced senescence in HAoECs. (A) The CCK-8 method was used to evaluate the effect of sodium nitrate on the proliferation of HAoECs. (B) Statistical analysis of cell proliferation at 96 h (A) . n = 3. (C) Representative images of SA-β-Gal-stained HAoECs. Bar = 100 μm. (D) Statistical analysis of the stained area in (C) . n = 5. (E) Representative images of γ-H2AX staining of HAoECs. Bar = 10 μm. (F) Statistical analysis of the DNA damage foci in (E) . n = 5. (G) QPCR analysis of the mRNA expression of the inflammatory factors IL-1β, IL-6, IL-8, and TNF-α in HAoECs. n = 3. (H) QPCR was used to detect the relative expression levels of the cell cycle-related genes p53, p21, p16 and Rb in HAoECs. n = 3. (I) Western blotting was used to detect the expression of P53, P21, and RB in HAoECs. n = 3 (J) Quantitative analysis of the results in (I) . *p < 0.05, ***p < 0.001, vs. the control group; # p < 0.05, ## p < 0.01, ### p < 0.001, vs. H 2 O 2 group.

Article Snippet: Human aortic endothelial cells (HAoECs) were purchased from ATCC (United States).

Techniques: CCK-8 Assay, Staining, Expressing, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Sodium nitrate regulates senescence accompanied by aortic atherosclerosis in ApoE −/− mice through the miR-34a/FGF-21 axis

doi: 10.3389/fphar.2025.1562321

Figure Lengend Snippet: Effects of miR-34a and FGF21 in ApoE −/− mice and HAoECs. (A) Venn plots for GeneCards database search for senescence and AS-related miRNA. (B) Top 10 miRNAs with the highest correlation scores associated with senescence in the common 276 miRNAs. (C) QPCR was used to detect the expression level of miR-34a in ApoE −/− mice at 12 weeks and HAoECs at 24 h. n = 3. (D) QPCR analysis of FGF21 mRNA expression in ApoE −/− mice at 12 weeks and HAoECs at 24 h. n = 3. (E) QPCR was used to detect the expression levels of miR-34a in HAoECs after treatment with the miR-34a mimic or inhibitor at 24 h. n = 3. (F) QPCR analysis of FGF21 mRNA expression in HAoECs after treatment with the miR-34a mimic or inhibitor at 24 h. n = 3. (G) The CCK-8 method was used to evaluate the effect of the miR-34a mimic on the proliferation of HAoECs at 96 h. n = 3. (H) CCK-8 was used to detect the effect of the miR-34a inhibitor on the proliferation of HAoECs at 96 h. n = 3. (I) A representative image of SA-β-Gal-stained HAoECs after treatment with the miR-34a mimic or inhibitor at 24 h. n = 5. Bar = 100 μm. (J) Statistical analysis of the stained area in (I) . n = 5. (K) A representative image of γ-H2AX staining of HAoECs after treatment with the miR-34a mimic or inhibitor at 24 h. Bar = 20 μm. (L) Statistical analysis of the number of DNA damage foci in (K) . n = 5. (M) QPCR analysis of the mRNA expression of the inflammatory factors IL-1β, IL-6, IL-8 and TNF-α in HAoECs after miR-34a mimic treatment at 24 h. n = 3. (N) QPCR analysis of the mRNA expression of the inflammatory factors IL-1β, IL-6, IL-8 and TNF-α in HAoECs after miR-34a inhibitor treatment at 24 h. n = 3. (O) QPCR was used to detect the relative expression levels of the cell cycle-related genes p53, p21, p16 and Rb in HAoECs after miR-34a mimic treatment at 24 h. n = 3. (P) QPCR was used to detect the relative expression levels of the cell cycle-related genes p53, p21, p16 and Rb in HAoECs in different treatment groups after miR-34a-5p inhibitor treatment at 24 h. n = 3. (Q) Western blotting was used to detect the protein expression of p53, p21, and Rb in HAoECs after miR-34a mimic or inhibitor treatment at 24 h. n = 3. (R) Quantitative analysis of the results in (Q) . *p < 0.05, **p < 0.01, ***p < 0.001, vs. the NFD group or the control group or mimic-NC; ## p < 0.01, ### p < 0.001, vs. HFD group or H 2 O 2 group or inhibitor-NC.

Article Snippet: Human aortic endothelial cells (HAoECs) were purchased from ATCC (United States).

Techniques: Expressing, CCK-8 Assay, Staining, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Sodium nitrate regulates senescence accompanied by aortic atherosclerosis in ApoE −/− mice through the miR-34a/FGF-21 axis

doi: 10.3389/fphar.2025.1562321

Figure Lengend Snippet: Effect of FGF21 on the senescence of HAoECs. (A) A CCK-8 kit was used to detect the effect of rhFGF21 on the proliferation of HAoECs at 48 h. (B) Statistical analysis of the HAoEC proliferation ability at 48 h (A) . n = 3. (C) Representative images of SA-β-Gal-stained HAoECs after treatment with the miRNA-34a mimic or rhFGF21 at 24 h. Bar = 100 μm. (D) Statistical analysis of the stained area in (C) . n = 5. (E) Representative images of γ-H2AX staining of HAoECs after miRNA-34a mimic and rhFGF21 treatment at 24 h. Bar = 20 μm. (F) Statistical analysis of the number of DNA damage foci in (E) . n = 5. (G) QPCR analysis of the mRNA expression of the inflammatory factors IL-1β, IL-6, IL-8 and TNF-α in HAoECs from different treatment groups at 24 h. n = 3. (H) QPCR was used to detect the relative mRNA expression of p53, p21, p16 and Rb in HAoECs after treatment with the miRNA-34a mimic or rhFGF21 at 24 h n = 3. (I) Western blotting was used to detect the protein expression of P53, P21, and RB in HAoECs after treatment with the miRNA-34a mimic or rhFGF21 at 24 h n = 3. (J) Quantitative analysis of the results in (I) . *p < 0.05, **p < 0.01, ***p < 0.001, vs. the mimic-NC group; # p < 0.05, ### p < 0.001, vs. the mimic group.

Article Snippet: Human aortic endothelial cells (HAoECs) were purchased from ATCC (United States).

Techniques: CCK-8 Assay, Staining, Expressing, Western Blot

Journal: PLOS ONE

Article Title: Hydrogen sulfide donor activates AKT-eNOS signaling and promotes lymphatic vessel formation

doi: 10.1371/journal.pone.0292663

Figure Lengend Snippet: (A) Human dermal lymphatic endothelial cells (LECs) were treated with PBS (vehicle) or various concentrations of NaHS (10–50 μM) for 48 h and cell proliferation determined using WST-1 assay. The data are representative of four independent experiments conducted at least in duplicate. (B) LECs plated on coverslips were treated with or without NaHS (30 μM) for 24 hours. Cells were immunostained for Ki67 (red) and nuclei counterstained with DAPI (blue). Microscopic images of five random fields were captured. Representative images are shown. Scale bar 50 μm. The bar graph shows the percentage of Ki67-positive nuclei ( n = 4). (C) Representative images of LEC migration after treatment with vehicle or NaHS (30 μM, 24h). The bar diagram shows the number of migrated cells ( n = 4). (D) LECs were seeded in wells of a Matrigel-coated-96-well plate in basal media (0.5% FBS) containing vehicle or NaHS (30 μM), and tube formation was investigated after 6 h. Representative images of tube formation are shown. Scale bar 1000 μm. Images of 3 random fields were captured, and the number of loops and branching points was quantified ( n = 4). Statistical analyses were performed using one-way ANOVA with Dunnett multiple comparisons test (A) , and two-tailed unpaired student t test (B-D) . Data represent mean ± SEM. * p < 0.05, ** p < 0.01, and **** p < 0.0001.

Article Snippet: In order to validate the identity of these cells, we performed quantitative real-time PCR (qRT-PCR) experiments to determine the mRNA expression of key LEC markers, FLT4 (VEGFR3) , LYVE-1 and podoplanin ( PDPN ) in these LECs and compared them with human aortic endothelial cells (HAoEC, Lonza) ( ).

Techniques: WST-1 Assay, Migration, Two Tailed Test