Chemical Profile, Antioxidant, Anti-Inflammatory, and Anti-Cancer Effects of Italian Salvia rosmarinus Spenn. Methanol Leaves Extracts

In this study, we evaluated and compared the chemical composition, the antioxidant,anti-inflammatory, and anti-proliferative effects of four methanol extracts (R1-R4), of Salvia rosmarinusSpenn. in two different sites of Southern Italy obtained by maceration or ultrasound-assistedextraction. Extracts of S. rosmarinus collected on the Ionian coast are indicated with the abbreviations R1(maceration) and R2 (ultrasound-assisted extraction). Extracts of S. rosmarinus collected on the Tyrrheniancoast are indicated with the abbreviations R3 (maceration) and R4 (ultrasound-assisted extraction).The chemical composition was analyzed using High Pressure liquid chromatography-Diod-Arraydetection-Electrospray ionization-Quadrupole-Mass Spectroscopy (HPLC-DAD-ESI-Q-MS).The antioxidant activity was analyzed by 2,20-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)(ABTS) 2,2-diphenyl-1-picrylhydrazyl (DPPH), -carotene bleaching, and Ferric Reducing AntioxidantPower (FRAP) assays. Antioxidant features were also assessed in lipopolysaccharide (LPS)-stimulatedRAW-264.7 murine macrophages, evaluating Reactive Oxygen Species (ROS) production; in the sameexperimental model, the anti-inflammatory activity of the extracts was investigated. Interestingly,all extracts displayed antioxidant and anti-inflammatory properties. They exhibited significativenitrite production inhibitory activity, whith IC50 values ranging from 3.46 to 5.53 g/mL, withoutimpairing cell viability. The anti-inflammatory activity was also investigated byWestern Blotting andimmunofluorescence assay, highlighting the R3 and R4 extracts ability to reduce NF-B translocation,as well as to disrupt the MAPKs signaling pathway. Extracts exhibited both potential anti-proliferativeactivity on breast cancer cells, inducing apoptosis, without aecting non-tumorigenic cells, and theability to inhibit MDA-MB-231 cells' motility. Finally, the rosemary extracts treatment significantlyreduced the power of conditioned media, from MCF-7 or MDA-MB-231 cells to induce nitriteproduction on RAW 264.7 cells, confirming their promising anti-inflammatory activity.