Table 1.
Effect of SphK, SMase, and S1P Lyase Inhibition and S1P Addition on MMP-9, MMP-2, and TIMP-1 Expression in H9c2 Cellsa
| Control | iSK | GW4869 | THI | S1P | |
|---|---|---|---|---|---|
| MMP-9 | |||||
| Ct | 29.66 | 24.1 | 24.6 | 24.47 | 25.02 |
| ΔCt | 7.64 ± 0.8 | 6.76 ± 0.6 | 6.72 ± 0.7 | 5.57 ± 0.7 | 7.25 ± 0.7 |
| MMP-2 | |||||
| Ct | 33.09 | 27.7 | 28.47 | 28.05 | 28.09 |
| ΔCt | 11.07 ± 1.0 | 10.38 ± 1.2 | 10.59 ± 1.0 | 9.15 ± 1.2 | 10.32 ± 1.1 |
| TIMP-1 | |||||
| Ct | 31.75 | 25.74 | 26.61 | 26.4 | 25.87 |
| ΔCt | 10.5 ± 1.1 | 9.35 ± 1.0 | 9.18 ± 1.1 | 9.33 ± 1.1 | 9.21 ± 1.0 |
a
MMP-9, MMP-2, and TIMP-1 expression was evaluated from RNA extracted from untreated H9c2 (control) cells and cells treated for 30 minutes with compound II (iSK, 5μM), GW9846 (10μM), THI (6.5μM), or S1P (1μM). Total RNA was reverse transcribed and real-time PCR amplification performed as described in Materials and Methods. ΔCt values are reported as mean ± SEM for independent experiments performed in duplicate. Significance of differences was tested by one-way ANOVA, and no significant change was observed in the presence of inhibitors vs control.