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. 2021 Sep 8;12:754976. doi: 10.3389/fphar.2021.754976

FIGURE 4.

FIGURE 4

FA attenuates TGF-β-induced HSC activation and fibrotic responses in LX-2 cells. (A) A CCK-8 assay. After pre-treated with FA at different concentrations for 1 h, LX-2 cells were treated with TGF-β (5 ng/ml) for another 24 h (B and C) or 6 h (E). (B) Relative mRNA levels of Tgfb1, Acta2, Fn, Col1a1 and H19 were determined by qPCR and normalized using Hprt1 as an internal control. (C) Representative collagen gel images from three independent experiments were shown. (D) LX-2 cells were treated with 25 μM FA at different time points or different concentrations of FA for 24 h. (F) After treated with CC (2 μM) or FA (25 μM) or both, LX-2 cells were administrated with TGF-β for another 6 h (D-F) Representative immunoblots against p-AMPK, t-AMPK, p-LKB1, t-LKB1, p-SMAD2, p-SMAD3, t-SMAD3, p-ERK1/2, t-ERK1/2 and β-ACTIN were shown. (G) The pathway of AMPK, TGF-β and SMADs involved in HSC proliferation. Statistical significance: **p < 0.01, ***p < 0.001, compared with control group; ## p < 0.01, ### p < 0.001, compared with TGF-β group. One-way ANOVA with Tukey’s post-hoc tests (n = 3).