Unveiling the in vitro activity of extracted Euphorbia trigona via Supercritical Fluid Extraction against pathogenic yeasts, obesity, cancer, and its wound healing properties

Abstract

Natural products of plant origin are being explored as safe alternatives for illness management. Their extraction processes play a crucial role in determining their phytochemical and pharmacological properties. In this context, Euphorbia trigona was extracted using Supercritical Fluid Extraction with CO₂ (SFE-CO₂) at two operating temperatures: 20 °C and 40 °C. Phytochemical characterization was performed via HPLC, along with anti-yeast evaluation using the well diffusion method, anticancer assessment using the MTT assay, wound healing analysis via the scratch assay, and anti-obesity evaluation through the lipase assay of the E. trigona extract. The results indicated that SFE-CO₂ at 40 °C extracted a greater quantity (0.198 g) of E. trigona than SFE-CO₂ at 20 °C (0.156 g). Several compounds, such as rosmarinic acid, gallic acid, daidzein, ellagic acid, naringenin, and ferulic acid, were identified at high concentrations of 10,034.29, 1,800.33, 750.22, 748.11, 462.15, and 207.05 µg/mL, respectively, in the E. trigona extract obtained using SFE-CO₂ at 40 °C, compared to the extract obtained using SFE-CO₂ at 20 °C. High inhibition zones of 24 ± 1.5, 24 ± 0.5, and 23 ± 0.33 mm were recorded against C. albicans, C. tropicalis, and G. candidum, respectively, using the extract from SFE-CO₂ at 40 °C, compared to the inhibition zones of 24 ± 1.5, 24 ± 0.5, and 23 ± 0.33 mm obtained from the extract using SFE-CO₂ at 20 °C. Moreover, the extract from SFE-CO₂ at 40 °C exhibited lower MIC and MFC values against the tested yeasts compared to the efficacy of the extract from SFE-CO₂ at 20 °C. The ultrastructure of the examined yeasts was severely affected by the extract from SFE-CO₂ at 40 °C. A lower IC₅₀ (98.87 ± 1.26 µg/mL) was recorded for the extract from SFE-CO₂ at 40 °C compared to the IC₅₀ (333.87 ± 1.8 µg/mL) of the extract from SFE-CO₂ at 20 °C against cancer cells (A431). The wound closure level was 84.08% using the extract from SFE-CO₂ at 40 °C, while it was 71.27% using the extract from SFE-CO₂ at 20 °C. Lipase was inhibited by the extract obtained via SFE-CO₂ at 40 °C and 20 °C, with IC₅₀ values of 15.77 and 28.14 µg/mL, respectively. Molecular docking indicated that rosmarinic acid is a suitable inhibitor for the tested yeasts.

Keywords: Euphorbia trigona; Cancer; Molecular docking; Rosmarinic acid; Wound healing; Yeasts.

Fig. 1

HPLC Chromatograms of separated compounds from E. trigona extract via SFE-CO₂ at 20 °C (A) and E. trigona extract via SFE-CO₂ at 40 °C (B)

Fig. 2

Anti-yeast properties of E. trigona extract at different temperatures of extraction. SFE-CO₂ at 20 °C (A), SFE-CO₂ at 40 °C (B), standard antifungal (S), and Negative control (NC)

Fig. 3

Image analysis of treated yeasts using TEM. Cell wall (CW), cytoplasm (Cy), cell membrane (CM), rupture cell wall (RCW), mitochondria (M), Vacuole (V) and Nucleus (N)

Fig. 4

Anticancer properties of E. trigona extract at different temperatures of extraction against A431 cells. SFE-CO₂ at 20 °C (left figure), SFE-CO₂ at 40 °C (right figure)

Fig. 5

Effect of E. trigona extract on the wound healing via Scratch assay. HFB4 cells at 0 h (A) and 48 h (B) without treatment compared to treated cells by extract via SFE-CO₂ at 20 °C (C) and SFE-CO₂ at 40 °C (D) after 48 h

Fig. 6

Effect of E. trigona extract and orlistat on lipase. Extract via SFE-CO₂ at 20 °C (T2) and SFE-CO₂ at 40 °C (T1) after 48 h

Fig. 7

2D and 3D graphs show the interaction among rosmarinic acid and active sites of C. albicans (PDB ID: 1ZAP) protein (A), rosmarinic acid and active sites of C. tropicalis (PDB ID: 6ZD6) protein (B), and rosmarinic acid and active sites of G. candidum (PDB ID: 6ISV) protein (C)