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. 2021 Aug 30;11:17343. doi: 10.1038/s41598-021-96761-2

Figure 1.

Figure 1

Protocatechuic acid reduces isoproterenol-induced cardiomyocyte hypertrophy in vitro. (A) H9c2 cells were treated with protocatechuic acid (0, 1, 10, 100 μM) for 24 h as indicated and cell viability was determined using the MTT assay (n = 8). NS indicates not significant. (B,C) H9c2 cells were serum-starved overnight and then treated with vehicle or protocatechuic acid in the presence or absence of isoproterenol (10 μM) for 24 h. Cell size was quantified by measuring the cell surface area of Alexa Fluor 488 phalloidin-stained cells (n = 40 cells). Scale bar = 50 μm. ***P < 0.001; ###P < 0.001. (D‒H) H9c2 cells were treated with isoproterenol (10 μM) in the presence or absence of protocatechuic acid (10 μM) for 6 h. The mRNA expression levels of Nppa (D), Nppb (E), and Myh7 (F) were determined by RT-PCR and normalized to Gapdh (n = 10). **P < 0.01 and ***P < 0.001; #P < 0.05 and ###P < 0.001. (G) Representative western blot images are shown; β-actin was used as a loading control. (H) Quantification of BNP protein levels (n = 6). ***P < 0.001; ###P < 0.001. ISO, isoproterenol; PCA, protocatechuic acid. Data are presented as the mean ± S.E. Statistical analysis: one-way ANOVA followed by Bonferroni post hoc tests. Graphs were prepared using GraphPad 5.0.