Panoply™ Human ACVR2A Over-expressing Stable Cell Line

Nonalcoholic steatohepatitis (NASH) is an increasingly common chronic liver disease, with liver fibrosis as its primary pathological change. The transforming growth factor β (TGF-β)/mall receptor antagonist Smad signaling pathway is a major factor in liver fibrosis. Although previous studies have suggested that echinacoside (Ech) has anti-fibrotic effects on liver fibrosis, the underlying cellular and molecular mechanisms remain unclear. Here, researchers investigated the anti-fibrotic properties of Ech in vivo and in vitro. Cell viability and scratch/wound assays demonstrated that Ech significantly inhibited the proliferation, migration, and activation of human hepatic stellate cells (LX-2). In mice with high-fat diet-induced liver fibrosis, Ech treatment attenuated liver injury, inflammation, and the progression of fibrosis. Furthermore, transcriptome analysis and subsequent functional validation demonstrated that Ech exerted its anti-fibrotic effects through the TGF-β1/Smad signaling pathway mediated by activin receptor type 2A (ACVR2A). Finally, by inhibiting and inducing ACVR2A expression in LX-2 cells, the researchers demonstrated that ACVR2A is an important target for liver fibrosis.

To determine whether the anti-fibrotic effects of Ech are dependent on ACVR2A inhibition, researchers generated ACVR2A-shRNA and ACVR2A-overexpressing LX-2 cells. Histological examination revealed that ACVR2A knockdown alleviated fibrosis similar to Ech treatment, while ACVR2A overexpression abolished the anti-fibrotic effects of Ech (Figures 1D-G). Co-immunofluorescence analysis of ACVR2A-shRNA or ACVR2A overexpression with β-SMA further demonstrated that ACVR2A is primarily expressed in activated HSCs. ACVR2A knockdown significantly suppressed β-SMA expression, while ACVR2A overexpression increased β-SMA expression (Figure 1J). Furthermore, ACVR2A knockdown was associated with a decrease in HSC activation markers (ACVR2A, COL1A1, β-SMA, and the p-SMAD2/3:SMAD2/3 ratio). However, in ACVR2A-overexpressing cells, HSC activation markers were significantly increased, even exceeding the inhibitory effect of Ech, particularly Smad phosphorylation (Figure 1H, I). These results suggest that blocking the ACVR2A-Smad signaling pathway can alleviate TGF-β1-induced hepatocyte fibrosis. Notably, Ech-targeted blockade of ACVR2A can trigger HSC activation, representing an attractive therapeutic strategy for liver fibrosis.

Figure 1. Ech is dependent on ACVR2A to relieve TGF-β1-induced liver cell fibrosis. (Liang J, et al., 2024)