Medicinal effects of Ephedra foeminea aqueous extracts: Metabolomic characterization, biological evaluation, and molecular docking

Abstract

Ephedra foeminea Forssk. is a medicinal plant traditionally used across various cultures and recognized for its historical significance in herbal medicine. It has been used as an herbal infusion to treat multiple respiratory diseases, headaches, and nasal congestion, as well as to prevent and manage breast cancer. In order to evaluate the validity of this ancestral knowledge, we aimed to investigate hot aqueous extracts of E. foeminea branches and fruits firstly by using untargeted metabolomic analysis, characterizing the chemical profiles of hot aqueous extracts from E. foeminea branches and fruits through the use of nuclear magnetic resonance, gas chromatography-mass spectrometry, and liquid chromatography-mass spectrometry. Subsequently, two in vitro studies were conducted to assess the anticancer and antioxidant potentials of the extracts. Antioxidant activity was evaluated using a 2,2-diphenyl-1-picrylhydrazyl assay and a total phenolic content assay. Anticancer activity was evaluated by assessing cytotoxicity using the MTT assay on MCF-7 (human breast cancer) and HeLa (cervical cancer) cell lines. Additionally, molecular docking was performed to explore the interactions between compounds identified in E. foeminea and selected cancer-related proteins, as well as the main protease of SARS-CoV-2. The results revealed that the branch extract exhibited superior antioxidant activity compared to the fruit extract, which was associated with a higher phenolic content in the branch extract (49.5 ± 0.7 mg GAE/g). The fruit extract exhibited greater cytotoxicity against MCF-7 cells, suggesting potential anticancer activity. Molecular docking analysis identified henryoside, guajavarin, and neohancoside as the most active compounds with anticancer and antiviral properties. These findings support the traditional use of E. foeminea Forssk. and highlight its potential as a source of bioactive compounds for further research into therapeutic applications.

Fig 1. Images of E. foeminea (Arabic common name: Ala’andah) grown in its native habitat in the West Bank of Palestine.

Fig 2. Representative 1D ¹H NMR spectra of E. foeminea branches (red) and fruits (blue).

The upper left inset highlights the aromatic region (6.7–9.5 ppm), and the upper right inset highlights the methylene region (1.5–3 ppm).

Fig 3. Graphical representation of the mean ratios of integrated regions from the NMR spectra of E. foeminea branch and fruit extracts.

Fig 4. 1D ¹³C solid-state NMR spectra of E. foeminea branches (red) and fruits (blue).

Fig 5. Percentage ratios of integrated regions from ¹³C solid-state NMR spectra of E. foeminea branch and fruit powders.

Data are based on a single measurement performed for chemical fingerprinting purposes; therefore, statistical analysis was not applicable.

Fig 6. Percentage of DPPH scavenging activity of E. foeminea branch and fruit extracts.

Fig 7. Total phenolic content of E. foeminea branch and fruit extracts.

Fig 8. Viability of HeLa, MCF-7, and Hek 295 cell lines treated with varying concentrations of E. foeminea branch aqueous extract.

Fig 9. Viability of HeLa, MCF-7, and Hek 295 cell lines treated with varying concentrations of E. foeminea fruit aqueous extract.

Fig 10. 2D and 3D interaction profiles of the henryoside ligand with breast cancer protein (PDB ID: 1HK7), HeLa cancer protein (PDB ID: 6I2I), and SARS-CoV-2 main protease (PDB ID: 6LU7).