Evaluation of the hepatoprotective properties of Vitex Altissima bark extracts and isolated bioactive: In-vitro and In-vivo approach

Vitex altissima ( V. altissima ), an ethnopharmacologically valuable plant, is explicitly used as traditional medicine by local folks to cure liver-related disorders in Coorg. Hence, the present study aimed to evaluate the bark extract's hepatoprotective and antioxidant mechanisms and its isolated potent compound against CCl 4 -induced toxicity. Ethanolic bark extracts underwent phytochemical screening, LC-MS profiling and bioactivity-guided antioxidant assays, including DPPH and hydroxyl radical scavenging methods. Preparative HPLC enabled the isolation of three compounds, among which Isolate-1 demonstrated the most potent bioactivity and was structurally characterized as Albafuran C via FTIR, NMR, and LC-MS/MS analyses. In vitro hepatoprotection studies using HepG2 cells revealed that pretreatment with Isolate-1 significantly enhanced cell viability, suppressed CCl 4 -induced oxidative stress, and restored antioxidant enzyme levels (SOD, CAT). Flow cytometry confirmed reduced intracellular ROS and iNOS expression, supporting its antioxidative and anti-inflammatory potential. In vivo experiments in CCl 4 -induced Wistar rats demonstrated significant amelioration of hepatic markers (ALT, AST, ALP, bilirubin, and albumin) and restoration of liver histoarchitecture upon treatment with Isolate-1. Molecular analysis by RT-PCR and Western blotting revealed downregulation of NF-κB signaling and inflammatory mediators. These findings confirm the hepatoprotective mechanism of V. altissima , substantiating its traditional use and identifying Albafuran C as a promising phytotherapeutic agent. This study establishes a scientific rationale for further development of Albafuran C as a natural hepatoprotective compound targeting oxidative and inflammatory liver injury. • Vitex altisimma bark extracts were screened for hepatoprotective activity. • Albafuran C was isolated from the bark extract of Vitex altisimma. • Albafuran C exhibited hepatoprotective and antioxidative effect. • Albafuran also shown good anti-inflammatory activity on liver.