Ferulic Acid Alleviates Oxidative Stress-Induced Cardiomyocyte Injury by the Regulation of miR-499-5p/p21 Signal Cascade

Figure 3

Ferulic acid protected cardiomyocyte from H₂O₂-induced cell injury by regulating the miR-499-5p/p21 signaling pathway. H9c2 cells were pretreated with 50 μM ferulic acid for 4 hrs and then incubated with 200 μM H₂O₂ for 24 hrs. (a), (b), (c). and (d) Western blot and the average protein level data of p-p38, p38, Bcl-2, Bax, and c-caspase-3 in the Con, H₂O₂, H₂O₂ + FA, and FA groups. GAPDH was used as the protein loading control (cropped blots,n = 4) (cropped blots, n = 4). (e) The miR-499-5p levels were analyzed by real-time PCR in the Con, H₂O₂, H₂O₂ + FA, and FA groups (n = 4). (f) The miR-499-5p levels were analyzed by real-time PCR after 0–300 μM H₂O₂ treatment for 24 hrs (n = 4). (g) H9c2 cells were treated with 0–100 μM ferulic acid and with/without 200 μM H₂O₂ treatment for 24 (h) and miR-499-5p was determined by real-time PCR (n = 5). (h), (i) Western blot and the average data of the miR-499-5p target protein p21 in Con, H₂O₂, H₂O₂ + FA, and FA groups. GAPDH was used as the protein loading control (cropped blots, n = 4). (j) miR-499-5p levels in miRNA negative control (Nc), miR-499-5p mimics, miRNA inhibitor negative control (Nci), and miR-499-5p inhibitor-transfected H9c2 cells (n = 3). (k). (l) Western blot and the average data of the protein level of p21 in miRNA negative control (Nc), miR-499-5p mimics (miR-499-5p-m), miRNA inhibitor negative control (Nci), and miR-499-5p inhibitor (miR-499-5p-i)-transfected H9c2 cells (n = 3). ^∗, ^∗∗, and ^∗∗∗.

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