Comparison of Carotenoids for Their Antifibrogenic Effects in Hepatic Stellate Cells

Abstract

Hepatic stellate cells (HSC) have an important role in the development of liver fibrosis by producing extracellular matrix proteins when they are activated upon liver injury. We previously demonstrated that astaxanthin (ASTX), a xanthophyll carotenoid, attenuates HSC activation. The objective of this study was to compare the anti‐fibrogenic effects of ASTX with those of other common carotenoids. LX‐2 cells, a human HSC cell line, were treated with ASTX, lycopene, lutein (LT), zeaxanthin, or canthaxanthin, to measure messenger RNA (mRNA) and protein expression of pro‐fibrogenic genes. Pro‐fibrogenic gene expressions were also measured in ASTX‐ or LT‐treated primary mouse HSC. To determine the underlying mechanisms of the anti‐fibrogenic effect of ASTX and LT, SMA‐related and MAD‐related protein 3 (SMAD3) pathways and the accumulation of reactive oxygen species (ROS) were measured in LX‐2 cells. Among five carotenoids tested, ASTX and LT attenuated HSC activation in LX‐2 cells by reducing the mRNA and protein levels of pro‐fibrogenic genes, such as smooth muscle α actin and procollagen type I α1 (COL1A1). In addition, both ASTX and LT significantly decreased the expression of pro‐fibrogenic genes, including COL1A1, COL3A1, and COL6A1, in activated primary mouse HSC, with ASTX being more potent than LT. The anti‐fibrogenic effect of ASTX was mediated by inhibiting the phosphorylation of SMAD3 and cellular ROS accumulation, while LT only prevented the accumulation of ROS in LX‐2 cells. In conclusion, ASTX showed the most potent anti‐fibrogenic effect among the five carotenoids via inhibition of SMAD3 phosphorylation and cellular ROS accumulation while LT only prevented ROS levels in HSC.