Abstract
In this study, phytochemical characterization of Anethum graveolens extract, in-vitro, in-vivo and in-silico biological activities and synergistic and antagonistic effects of main components were investigated. Advanced analytical techniques such as LC-MS/MS and GC-MS were integrated with in-vitro, in-vivo and in-silico methodologies to provide valuable information on the therapeutic potential of the plant and its synergistic and antagonistic interactions, which are crucial for developing effective plant-based therapeutic agents. LC-MS/MS analysis revealed the presence of protecatechuic aldehyde, sesamol, caffeic acid, vannilin, p-coumaric acid, rutin and quercetin among the standards tested. Among the essential oils, γ-terpinene, limonene, γ-cadinene, dillapiole were detected in major proportions. The radical scavenging activity of A. graveolens extract was investigated by DPPH and superoxide anion scavenging tests. DPPH scavenging activity was observed in the range of 15.9-80.8% and superoxide anion scavenging activity in the range of 11.5-69.9%. The extract exhibited a broad spectrum activity against all tested microorganism species and the antimicrobial activity ranking among microorganisms was gram-positive > gram-negative > fungus. A. graveolens extract exhibited an inhibition zone of 15.2 ± 0.4 against E. coli and the main components tested individually exhibited a lower activity. And also MIC and MBC values obtained with four main components were higher compared to the extract.This indicates that the extract exhibits a stronger antimicrobial activity due to the cumulative and synergistic effects of the active ingredients in the content. The most prominent synergistic effect was found in anti-genotoxic activity. While the most common fragment rate among the chromosomal aberrations induced by NaN(3) decreased by 73.3% with the 20 mg/L extract application, the fragment ratio decreased by 37.3%, 35.3%, 31.5% and 28.07% with the caffeic acid, rutin, dillapiole and terpinen applications, respectively. These results suggest that the ingredients have a strong synergistic effect. In molecular docking studies the major components of A. graveolens were found to interact with aquaporin and DnaK ligands by hydrogen bonds, pi-aclyl, amide-Pi stacked, and pi-sigma bonds. The interaction of these proteins with the main components may cause disruptions in microbial metabolism and an indirect antimicrobial activity.