Liver fibrosis results from chronic liver injury and is characterized by the accumulation of extracellular matrix in excess driven by hepatic stellate cells (HSCs) activation. Chrysin (CHR) is a natural flavonoid that is limited by its low solubility to exert its anti-inflammatory, antioxidant and anti-fibrotic properties. The aim of this study was to investigate the biocompatibility of CHR complexes with two cyclodextrins (CDs)-(2-hydroxypropyl)-β-cyclodextrin (HPBCD) and random methyl-β-cyclodextrin (RAMEB), and their potential to induce anti-inflammatory, antioxidant and anti-fibrotic effects. Biocompatibility of the complexes was evaluated on Huh7 and LX2 cell lines: MTT and Live/Dead tests indicated the cell viability and an LDH test showed the cytotoxicity. Immunohistochemical staining of Nuclear Factor Kappa B (NF-κB) nuclear translocation was performed to evaluate the anti-inflammatory effect of the complexes. Oxygen Radical Absorbance assay, Superoxide Dismutase activity and Glutathione Peroxidase (GPx) assays indicated the antioxidant properties of the chrysin complexes. Finally, the complexes' anti-fibrotic potential was evaluated at the protein and gene level of α-sma. In HSCs, CDs induced higher cytotoxicity correlated with lower cell viability than CHR-CD. The 1:1 CHR-RAMEB pretreatment avoided p65 translocation. The 1:2 CHR-RAMEB complex increased ORAC values, improved SOD activity and produced the highest stimulation of GPx activity. CHR-RAMEB reduced α-sma expression at lower concentration than CHR-HPBCD, proving to be more efficient. In conclusion, both CHR-CD complexes proved to be biocompatible, but CHR-RAMEB showed improved anti-inflammatory, antioxidant and anti-fibrotic effects that could recommend its further use in liver fibrosis treatment.
Keywords: anti-fibrotic; anti-inflammatory; antioxidant; biocompatibility; chrysin; cyclodextrins; hepatic stellate cells; liver fibrosis.