Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025 Oct 23;15(1):37154.
doi: 10.1038/s41598-025-21041-2.

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

Mosad A Ghareeb  1 , Boulanouar Bakchiche  2   3 , Yacine Aouiffat  2   3 , Tarek Aboushousha  4 , Mohamed Marzouk  5 , Hend Okasha  6
Affiliations

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

Mosad A Ghareeb et al. Sci Rep. .

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.

Keywords: Haloxylon scoparium; In silico study; Good health and well-being; Hepatocellular carcinoma; LC/ESI-MS/MS; Polyphenols; TNF-α.

PubMed Disclaimer

Conflict of interest statement

Declarations. Compliance with ethical standards: All procedures in this study, including handling, anesthesia, and sacrifice, adhered to the ethical guidelines approved by the Ethical Committee of the Federal Legislation and the National Institutes of Health Guidelines in the United States. All animal experiments were carried out under Institutional Ethical Committee rules for the care and use of experimental animals, which were authorized by Theodor Bilharz Research Institute’s Animal Ethics Committee in Giza, Egypt (FWA 00010609). All methods are documented in line with the ARRIVE guidelines for reporting animal experiments. ( https://arriveguidelines.org ). Competing interests: The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
(a) Percent of surviving mice at the end of the experiment (12 weeks). Results showed that H. scoparium post-treatment protected mice from death, and its dose was safe, as found in the H. scoparium group. (b) Effect of the H. scoparium extract on the b. w. of mice with HCC induced by DEN.
Fig. 2
Fig. 2
Effect of the H. scoparium extract on the liver function in DEN-induced HCC in mice.
Fig. 3
Fig. 3
Relative quantification of inflammatory marker (TNF-α), tumor markers (AFP and BCl-2), and apoptotic markers (BAX, Caspase-3, and Cas-8) in HCC-induced mice.
Fig. 4
Fig. 4
Histopathology of the liver isolated from mice with DEN-induced HCC.
Fig. 5
Fig. 5
Immunohistochemistry of Caspase-3 protein detection (X400). (a) Section in the liver of Normal mice showing predominantly sinusoidal expression of Caspase-3 with moderate intensity. (b) Section in the liver of DEN-treated mice showing remarkable dense expression of Caspase-3 within hepatocytes (brownish discoloration). (c) Section in the liver of HS-treated mice showing focal mild expression of Caspase-3 within hepatocytes and sinusoids. (d): Section in the liver of HS + DEN-treated mice showing increased dense expression of Caspase-3 within hepatocytes.
Fig. 6
Fig. 6
TIC chromatogram from LC/ESI-MS/MS analysis of H. scoparium leaf extract.
Fig. 7
Fig. 7
3D binding interaction and Box plot representing the docking score (binding affinity) in kcal/mol of Quercetin triglycoside with the BCL-2 protein.
Fig. 8
Fig. 8
3D binding interaction and Box plot of Quercetin triglycoside with the BAX protein.
Fig. 9
Fig. 9
3D binding interaction and Box plot of Isorhamnetin triglycoside with Caspase-3.
Fig. 10
Fig. 10
3D binding interaction and Box plot of Quercetin triglycoside with Caspase-8.
Fig. 11
Fig. 11
3D binding interaction and Box plot of Isorhamnetin triglycoside with AFP.
Fig. 12
Fig. 12
3D binding interaction and Box plot of Isorhamnetin triglycoside with TNF-α.
See this image and copyright information in PMC

References

    1. Boucherit, H. & Benaradj, A. Therapeutic properties of the woody plant Haloxylon scoparium Pomel in the steppe region of Naâma (Algeria). Biodivers. Res. Conserv.72, 39–46 (2024).
    1. Amtaghri, S. & Eddouks, M. Comprehensive review on the genus Haloxylon: Pharmacological and phytochemical properties. Endocr. Metab. Immune Disord Drug Targets. 24, 1146–1160 (2024). - PubMed
    1. Purayil, F. T. et al. Transcriptome profiling of Haloxylon persicum (Bunge ex Boiss and buhse) an endangered plant species under peg-induced drought stress. Genes11, 640 (2020). - PMC - PubMed
    1. Bouaziz, A. et al. Antibacterial and antioxidant activities of Hammada scoparia extracts and its major purified alkaloids. S Afr. J. Bot.105, 89–96 (2016).
    1. Ebrahim, A. M., Idris, A. M., Alnajjar, A. O. & Michalke, B. Cr and Mn total, accessible species, and protein-fraction contents in plants used for traditional anti-diabetes treatment. J. Trace Elem. Med. Biol.62, 126645 (2020). - PubMed

MeSH terms