Authors
Nashaat N Mahmoud, Alsayed E Mekky, Ebrahim Saied, Faisal Alsenani, Hamed Alsufyani, Adnan Alharbi, Mohammad Y Alfaifi, Ali A Shatii, Islam Abdelhakim Seyiam, Mohammed Aufy
Published in
Scientific reports. Volume 16. Issue 1. Aug 10, 2026. Epub Aug 10, 2026.
Abstract
Echium angustifolium is traditionally valued for its therapeutic properties; yet comprehensive studies on its biological activities are limited. Aerial flowering parts were sequentially extracted with solvents of varying polarity. Extracts were analyzed for yield, phytochemical composition, and total phenolic and flavonoid contents and HPLC. LC-MS/MS analysis in negative ion mode was performed for detailed compound identification. Antioxidant activity was measured using the DPPH assay. Antibacterial activity against Gram-positive and Gram-negative bacteria was evaluated via agar diffusion and MIC/MBC methods. Anti-inflammatory potential was determined in vitro through RBC membrane stabilization (hemolysis) assay. Cytotoxicity and antiviral activity were examined using MTT and cell-based assays on HepG2 and Vero cells. The molecular mechanism of anticancer activity was investigated through qPCR analysis of apoptosis-related genes (Bax, Caspase-3, Caspase-9, and Bcl-2) in HepG2 cells. Antiviral efficacy against Hepatitis A virus (HAV) and Coxsackievirus B4 (CoxB4) was further validated by RT-qPCR quantification of viral RNA copy numbers. The ethanolic extract showed the highest yield (14.23%) among the tested solvents and richest phytochemical profile, with rutin as the predominant compound. LC-MS/MS identified 24 compounds including threonic acid, hypoxanthine, indole-3-carboxylic acid, N-methyllysine, 4-methyl-2-oxovaleric acid, 4-isopropylbenzoic acid, methionine, N-2-fluorenylacetamide, arachidic acid, N-methylanthranilic acid, carbocysteine, fructose-1,6-bisphosphate, lumazine, 5-aminoimidazole-4-carboxamide, and 3-isochromanone. It exhibited significant antioxidant activity (IC50 = 21.08 µg/mL) and antibacterial effects against both Gram-positive and Gram-negative bacteria. It exhibited in vitro anti-inflammatory effects via RBC membrane stabilization (95.5% inhibition at 1000 µg/mL) and dose-dependent cytotoxicity against HepG2 cells (IC50 = 94 µg/mL), while showing low toxicity toward Vero cells (IC50 = 632.24 µg/mL). Mechanistically, qPCR analysis revealed that the extract at IC50 concentration significantly upregulated pro-apoptotic Bax (4.21-fold), Caspase-3 (5.02-fold), and Caspase-9 (4.08-fold), while downregulating anti-apoptotic Bcl-2 (0.33-fold) compared to untreated controls, confirming apoptosis induction through the intrinsic mitochondrial pathway. Antiviral assays revealed significant inhibition of HAV and Coxsackievirus B4 replication at non-toxic concentrations. RT-qPCR quantification demonstrated that the extract at MNTC (250 µg/mL) reduced HAV and CoxB4 viral RNA copies by 50.6% (from 1.22 × 106 to 6.03 × 105 copies/mL for HAV; from 2.47 × 105 to 1.22 × 105 copies/mL for CoxB4), providing direct evidence of viral inhibition. The ethanolic extract of E. angustifolium exhibited multifunctional in vitro bioactivities, including antioxidant, antimicrobial, anti-inflammatory, anticancer, and antiviral effects. LC-MS/MS identified a diverse array of bioactive compounds, while mechanistic studies revealed apoptosis induction via the mitochondrial pathway (upregulation of Bax, Caspase-3, and Caspase-9; downregulation of Bcl-2) and direct antiviral activity through inhibition of viral replication (confirmed by RT-qPCR). These findings validate the traditional medicinal uses of E. angustifolium and highlight its potential as a promising source of bioactive compounds that warrant further in vivo and mechanistic investigations.
PMID:
42575926
Bibliographic data and abstract were imported from PubMed on 11 Aug 2026.
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