LC/ESI-MS/MS phytochemical profiling and apoptotic effect of Haloxylon scoparium leaf extract on hepatocellular carcinoma

Abstract

Abstract Haloxylon scoparium , a plant native to Moroccan Sahara, was investigated for its potential anticancer activity against hepatocellular carcinoma (HCC). The study aimed to evaluate the effects of its methanolic extract on HCC and to conduct detailed chemical analysis using LC-ESI-MS/MS. In vitro cytotoxicity was assessed using HepG2 liver cancer cell line. In vivo experiments involved inducing HCC in mice with diethylnitrosamine (DEN). The study monitored inflammatory (TNF-α), apoptotic (BAX, Caspase-3, Caspase-8), and oncogenic markers (AFP, Bcl-2) through blood and liver tissue analysis. Liver histopathology was also performed to evaluate tissue-level changes. Mice survival rates were 83.33% in the DEN group and 91.67% in the DEN/ H. scoparium group. Liver function markers (TBILR, ALP, AST) significantly decreased in the treatment group. TNF-α levels, elevated in DEN-only mice, were notably reduced after treatment. Oncogenic markers showed significant elevation in the DEN group but were decreased in the treatment group, whereas apoptotic markers were significantly elevated after treatment. Histopathology revealed more preserved liver architecture and scattered apoptotic foci in treated mice. Phytochemical profiling identified 27 compounds, including organic acids, phenolic acid derivatives, and flavonoids by LC/ESI-MS/MS. Molecular docking using AutoDock MGLTools 1.5.7 showed strong binding affinities of quercetin and isorhamnetin glycosides with cancer-related proteins (BCL-2, BAX, Caspases, AFP, TNF-α), supporting experimental results. 3D interaction models and box plots confirmed the stability and specificity of ligand–protein interactions. The study concludes that H. scoparium extract demonstrates promising multi-target anticancer potential and may serve as a valuable candidate for pharmaceutical development.