Table 2.
Triamterene inhibited the effects of lithocholic acid (LCA) on the mRNA levels of hypertrophic biomarkers and intracellular calcium ions, in addition to the quantified data from Fig. 1C,D
| Parameters | Control | Vehicle in High Glucose | LCA(5 × 10−6 M) in High Glucose | Triamterene(5 × 10−7M) + LCA(5 × 10−6 M) in High Glucose | Triamterene(1 × 10−6M) + LCA(5 × 10−6 M) in High Glucose |
|---|---|---|---|---|---|
| Cell size level (fold change) (n = 4) | 1.00 ± 0.10 | 3.51 ± 0.22** | 1.09 ± 0.14## | 1.94 ± 0.13*# | 3.81 ± 0.25** |
| Relative level of ANP/β-actin (n = 6) | 1.00 ± 0.00 | 1.87 ± 0.07** | 1.18 ± 0.09## | 1.48 ± 0.04*# | 1.94 ± 0.03** |
| Relative level of BNP/β-actin (n = 6) | 1.00 ± 0.00 | 1.85 ± 0.05** | 1.14 ± 0.05## | 1.52 ± 0.04*# | 1.75 ± 0.07** |
| Relative level of β-MHC/β-actin (n = 6) | 1.00 ± 0.00 | 1.85 ± 0.03** | 1.17 ± 0.05## | 1.49 ± 0.07*# | 1.93 ± 0.03** |
| [Ca2+](nM) (n = 6) | 168.19 ± 4.65 | 247.83 ± 6.47** | 175.03 ± 4.57## | 199.33 ± 7.20*# | 239.05 ± 8.16** |
| Ratio of TGR5/β-actin protein (n = 4) | 0.28 ± 0.03 | 0.58 ± 0.05* | 0.83 ± 0.07**# | 0.70 ± 0.09**# | 0.54 ± 0.02* |
| Ratio of Calcineurin/β-actin protein (n = 4) | 0.33 ± 0.06 | 0.86 ± 0.03** | 0.50 ± 0.04## | 0.67 ± 0.03*# | 0.84 ± 0.08** |
| Ratio of NFAT3/Histone H3 protein (n = 4) | 0.41 ± 0.06 | 0.78 ± 0.08** | 0.43 ± 0.08## | 0.61 ± 0.05*# | 0.76 ± 0.06** |
The changes in cell size shown in Fig. 1C were quantified for comparison in the first row. The cell size enhanced by high glucose (30 mM) in the H9c2 cells was reduced by lithocholic acid (LCA) at the indicated dose, and this effect was reversed by triamterene in a dose-related manner. Similarly, the increased mRNA levels of hypertrophic markers, including ANP, BNP, and β-MHC, by high glucose that was attenuated by LCA were also reversed by triamterene. The merits of LCA for the amelioration of cellular hypertrophy, which was supported by changes in hypertrophic signals of calcineurin and NFAT3, were reversed by triamterene as shown in the last two rows. Moreover, similar changes in the calcium levels are indicated in the middle of the table. Each value is shown as the mean ± SEM at the indicated sample size (n) per group. *P < 0.05 and **P < 0.01 vs the control. #P < 0.05 and ##P < 0.01 vs the vehicle-treated samples in high glucose (Vehicle in High Glucose).