Cyclopia intermedia E. Mey protects against ROS-induced liver injury in HepG2/C3A cells

Abstract

Drug-induced liver injury (DILI) represents a significant clinical problem for which no standard treatment currently exists. Most DILI mechanisms center on oxidative stress resulting from glutathione depletion, excessive ROS production, and subsequent cell death. Flavonoid-rich Cyclopia intermedia E. Mey (honeybush) is a commercially important African herbal tea and is traditionally promoted as a restorative treatment. Cytotoxicity evaluation of a fermented C. intermedia extract in C3A hepatocytes confirmed the absence of toxicity up to 200 µg/mL using quantitative fluorescence microscopy with Hoechst 33342-PI dual-labeling. DPPH, NO scavenging, and ORAC assays established a strong antioxidant potential. Antioxidant bioavailability, assessed using the CAPe assay, indicated significant uptake at 100 µg/mL (p < 0.005). C. intermedia extract decreased TBHP-induced oxidative stress in C3A cells and attenuated TBHP-induced changes in thiol group levels at 200 µg/mL (p < 0.005) as confirmed by CellROX® Orange and ThiolTracker™ Violet staining, respectively. Apoptotic cell death was significantly reduced by the extract from 50 µg/mL (p < 0.005) as determined by Annexin-V FITC staining. These results suggest that C. intermedia can function as a hepatoprotectant and has potential as a treatment against DILI.