AUTHOR=Babotă Mihai , Frumuzachi Oleg , Nicolescu Alexandru , Stojković Dejan , Soković Marina , Rocchetti Gabriele , Zhang Leilei , Lucini Luigi , Crișan Gianina , Mocan Andrei , Voștinaru Oliviu 

TITLE=Phenolic profile, in vitro antimicrobial and in vivo diuretic effects of endemic wild thyme Thymus comosus Heuff ex. Griseb. (Lamiaceae) from Romania

JOURNAL=Frontiers in Pharmacology

VOLUME=Volume 14 - 2023

YEAR=2023

URL=https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2023.1115117

DOI=10.3389/fphar.2023.1115117

ISSN=1663-9812

ABSTRACT=Thymus comosus Heuff ex. Griseb. (Lamiaceae) is a wild thyme species endemic for Romanian Carpathian areas, frequently collected as substitute for collective herbal product Serpylli herba, cited as antibacterial and diuretic remedy in traditional medicine. The present study aimed to evaluate the in vivo diuretic effect and in vitro antibacterial properties of three herbal preparations (infusion – TCI, tincture – TCT and an hydroethanolic extract prepared through an optimized ultrasound-assisted method – OpTC) obtained from the aerial parts of T. comosus Heuff ex. Griseb, also evaluating their comprehensive phenolic profile. 
In vivo diuretic effect was tested using Wistar rats treated orally with each herbal preparation (125 and 250 mg/kg dispersed in 25 mL/kg isotonic saline solution) and quantified based on cumulative urine output (mL), diuretic action (defined as the ratio of urine output in the test groups to urine output in the control group) and diuretic activity (defined as the ratio of urine output in the test groups to urine output of the reference group). Additionally, sodium and potassium excretion were monitored using a potentiometric method with selective electrodes. In vitro antibacterial and antifungal activities were assessed using p-iodonitrotetrazolium chloride assay against six bacterial strains and six fungal strains by monitoring minimum inhibitory concentration (MICs), minimum bactericidal concentrations (MBCs) and minimum fungicidal concentrations (MFCs). All the extracts exerted a mild diuretic action, TCT and OpTC inducing the most intense diuretic effect. Both herbal preparations produced a statistically significant, dose-dependent and gradual increase of the urine output, the effect being more intense at 24 h. In terms of antimicrobial activity, E. coli, Salmonella Typhimurium, Penicillium funiculosum and P. verrucosum var. cyclopium showed the greater sensitivity to the tested extracts, respectively. HRMS screening showed that the bioactive potential of T. comosus  herbal preparations was likely related to the higher amounts of phenolic acids (including rosmarinic acid), flavonoids (mainly flavones and derivatives) and other phenolics (such as different isomers of salvianolic acids) in their composition. Results support the ethnopharmacological evidence regarding the diuretic and antibacterial potentials of T. comosus, this study being the first one that assessed the aforementioned bioactivities for this species.