Raman imaging investigation of hepatic LX-2 cell reversion under different lipidic treatments

Abstract

Liver fibrosis resulting from chronic liver injury is characterized by increased extracellular matrix deposition and inflammation, which leads to excessive scar tissue formation. Targeting activated hepatic stellate cells (HSCs), which are the primary drivers of fibrogenesis, stands out as one of the most compelling therapeutic approaches in this regard. In a healthy liver, HSCs remain quiescent and store vitamin A in cytoplasmic lipid droplets. As a consequence of HSC activation and transdifferentiation to a proliferative myofibroblast-like state upon fibrotic stimuli, the distinctive phenotypic feature of the lipid droplets gets lost. While the reversal of activated HSCs is feasible, understanding the quiescent-like state following injury resolution is crucial for effective fibrosis treatment. This study explores the induced quiescent-like state of naïve immortalized human hepatic stellate (LX-2) cells when treated with soybean phospholipid that contains 75% phosphatidylcholine (S80). The lipid profile of the newly formed lipid droplets was analyzed using Raman imaging, which is a label-free technique well-suited for lipidomics. Results indicate the presence of distinct lipid profiles despite maintaining a quiescent-like state, suggesting that diverse mechanisms govern the active-to-inactive state transition. Additionally, our findings support the fact that each hepatic cell state is composed of heterogeneous subpopulations. This emphasizes the complexity of liver fibrosis and highlights the need for a comprehensive understanding of cellular states to develop targeted therapies.