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