Marrubium vulgare (Lamiaceae) is a plant which has long been known and used in traditional medicine for various purposes. However, few recent studies have documented its chemical composition and biological properties. The present study investigated the phytochemical composition of horehound, as well as its protective, antioxidant, and antimicrobial potential. GC-MS analysis revealed that the major components of horehound essential oil are E-caryophyllene (35.7%), germacrene D (25.2%), and bicyclogermacrene (10.6%). The biological activity of horehound hydroethanolic herb extract derives from multiple chemical compounds, including polyphenols (55.72 mg/mL), flavonoids (11.01 mg/mL), phenolic acids (4.33 mg/mL), and tannins (4.46 mg/mL). Chromatographic analyses of the extract identified 12 phenolic compounds, of which ferulic acid, catechin, quercetin, protocatechuic acid, rutin, and syringic acid (35.42, 24.69, 20.65, 18.70, 14.46, and 12.69 mg/mL, respectively) were the main constituents. Its DPPH radical scavenging ability was 68.29%, while its antioxidant properties, determined by the FRAP method, were at the level of 1.22 mmol/L. Moreover, M. vulgare extract decreased the level of intracellular reactive oxygen species in the fibroblasts and keratinocytes in vitro, achieving the strongest antioxidant effect at a concentration of 2.5% in the case of both types of skin cells. Extracts from the horehound herb showed significant antimicrobial and anti-biofilm activity, confirming the plant’s potential in therapeutic applications against various microbial pathogens (gram-positive and gram-negative bacteria and fungi). The research results demonstrate the protective effect of horehound extract on the viability of both fibroblasts and keratinocytes in vitro. To sum up, M. vulgare, as a valuable natural material with high preventive and therapeutic effectiveness, is a potential candidate for new applications in the pharmaceutical and cosmetics industries.
