Alleviation of hepatic ischemia-reperfusion injury by ginsenoside Rg5 and deoxyandrographolide through inhibition of SLC7A11/GPX4-mediated ferroptosis.

Shenfu Injection is a traditional Chinese medicine formulation widely used for cardiovascular and ischemic disorders. This study investigated the protective effects and underlying mechanisms of its bioactive constituents, ginsenoside Rg5 (Rg5) and deoxyandrographolide (DA), against hepatic ischemia-reperfusion injury (HIRI). Mouse HIRI models and oxygen-glucose deprivation/reoxygenation (OGD/R)-treated AML12 cells were used for in vivo and in vitro studies, respectively. Combined Rg5 and DA treatment alleviated hepatic histopathological injury, reduced serum alanine aminotransferase and aspartate aminotransferase levels, restored antioxidant capacity, and decreased inflammatory responses and hepatocyte apoptosis. Transcriptomic and metabolomic analyses indicated that ferroptosis, glutathione metabolism, and lipid peroxidation-related pathways were involved in the protective effects of Rg5 and DA. Molecular docking and molecular dynamics simulations predicted interactions between Rg5 or DA and the ferroptosis-related proteins SLC7A11 and GPX4. Moreover, combined treatment increased SLC7A11 and GPX4 expression at both the mRNA and protein levels and decreased PTGS2 mRNA expression in vivo and in vitro. These findings suggest that Rg5 and DA protect against HIRI, by regulating SLC7A11/GPX4-associated ferroptosis, while also attenuating inflammation and hepatocyte apoptosis.