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Q: Non-alcoholic steatohepatitis (NASH) is a subset of non-alcoholic fatty liver disease, the most common chronic liver disease in the U.S. Fibrosis, a common feature of NASH, results from the dysregulation of fibrogenesis in hepatic stellate cells (HSCs). In this study, we investigated whether astaxanthin (ASTX), a xanthophyll carotenoid, can inhibit fibrogenic effects of transforming growth factor 1 (TGF1), a key fibrogenic cytokine, in HSCs.', 'Reactive oxygen species (ROS) accumulation was measured in LX-2, an immortalized human HSC cell line. Quantitative realtime PCR, Western blot, immunocytochemical analysis, and in-cell Western blot were performed to determine mRNA and protein of fibrogenic genes, and the activation of Smad3 in TGF1-activated LX-2 cells and primary mouse HSCs.', 'In LX-2 cells, ROS accumulation induced by tert-butyl hydrogen peroxide and TGF1 was abolished by ASTX. ASTX significantly decreased TGF1-induced -smooth muscle actin (-SMA) and procollagen type 1, alpha 1 (Col1A1) mRNA as well as -SMA protein levels. Knockdown of Smad3 showed the significant role of Smad3 in the expression of -SMA and Col1A1, but not TGF1, in LX-2 cells. ASTX attenuated TGF1-induced Smad3 phosphorylation and nuclear translocation with a concomitant inhibition of Smad3, Smad7, TGF receptor I (TRI), and TRII expression. The inhibitory effect of ASTX on HSC activation was confirmed in primary mouse HSCs as evidenced by decreased mRNA and protein levels of -SMA during activation.
A:
Does astaxanthin prevent TGF1-induced pro-fibrogenic gene expression by inhibiting Smad3 activation in hepatic stellate cells?