Figure 3.

6-shogaol treatment significantly suppressed TGF-β-induced fibrotic responses in cultured cardiac fibroblasts. Primary cultured cardiac fibroblasts derived from neonatal SD rats were pretreated with 6-shogaol (0.3 or 1 μM), then stimulated with TGF-β (10 ng/mL) for 24 h or 48 h. (A) [3H]-L-proline incorporation that was induced by TGF-β was investigated by liquid scintillation counter. (B–D) At 24 h after stimulation, quantitative RT-PCR was used to examine the mRNA levels of fibrosis-related gene transcription of α-SMA, periostin, and Col1a1. WCL was extracted from cardiac fibroblasts after 24 h of TGF-β stimulation. (E) Representative image of Western blotting and (F) quantified α-SMA levels. All panels show the mean ± SEM of four individual experiments.