Berberis vulgaris extract/β-cyclodextrin complex increases protection of hepatic cells via suppression of apoptosis and lipogenesis pathways

Berberis vulgaris (Bv) is well known worldwide for its healing properties. However, limited information is available concerning its mechanism of action and the increased hepatoprotective activity of formulated extracts. This study evaluated the protective effect of Bv bark extract against CCl(4)-induced cytotoxicity in Huh7 cells, as well whether β-cyclodextrin complexation of the extract resulted in increased hepatoprotective effects. Huh7 cells were incubated for 48 h with 5, 7.5 and 10 µg/ml of unformulated or formulated Bv extract alone and in co-treatment with CCl(4). The effects on Huh7 cell growth and apoptosis were evaluated by MTT assay, caspase-3/7 activity and caspase-3 expression, whereas fatty acid changes were investigated by Oil red O staining and the detection of peroxisome proliferator-activated receptor-γ (PPARγ) expression using immunofluorescence. Ultrastructural alterations were observed by electron microscopy. The MTT assay showed that co-exposure to CCl(4) and 7.5 µg/ml formulated extract led to a 1.25-fold increase in cell viability compared with the non-formulated extract. Caspase-3/7 activity decreased by 50% and 70% following co-treatment with unformulated or formulated extract, compared with that in cells treated with CCl(4) alone. Furthermore, hepatocyte ultrastructure was protected from CCl(4)-induced injury in the two co-treated groups, intracytoplasmic lipid accumulation decreased significantly and PPARγ expression was restored, in comparison with CCl(4)-treated cells alone. Formulated and unformulated extracts were efficient against the anti-proliferative and pro-apoptotic actions of CCl(4) through suppression of CCl(4)-induced caspase-3 activation and lipid accumulation. The protective effect of the formulated extract was more pronounced than that of the unformulated one, which may be due to its increased solubility.