Fig. 4
The antifibrotic effect of 2ccPA on SSc skin fibroblasts. a SSc skin fibroblasts (n = 5) were incubated with 0–10 μM 2ccPA for 48 h. The expression of COL1A1, COL1A2, CTGF, and ACTA2 mRNA was calculated by qPCR. b Time course expression of profibrotic markers in SSc skin fibroblasts (n = 5) treated with 2ccPA. SSc skin fibroblasts were incubated with 10 μM 2ccPA for 6–48 h. The expression of COL1A1, COL1A2, ACTA2, fibronectin (FN), and TGF-β1 mRNA was calculated by qPCR, and the amount of expression was compared with that of no treatment group. c The expression of type I collagen and CCN2 in cell lysates of SSc skin fibroblasts (n = 3) treated with 2ccPA. SSc skin fibroblasts were incubated with 0–10 μM 2ccPA for 48 h. The protein expression of cell lysates was detected by Western blotting. To calculate the levels of procollagen type I in the medium, SSc skin fibroblasts (n = 8) were incubated with 0–10 μM 2ccPA for 72 h and cultured medium were assessed using a commercially available enzyme immunoassay kit. d SSc skin fibroblasts (n = 8) were incubated with 10 μM 2ccPA for 24 h. PGE2 and HGF levels in the cultured medium were calculated by commercially available enzyme-linked immunosorbent assay kits. The bars represent as median with IQR in a–c. Data are the representative of three independent experiments in a–c. Bars represent the mean ± standard deviation (SD) in d. *p < 0.05