Abstract
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 rosmarinus Spenn. in two different sites of Southern Italy obtained by maceration or ultrasound-assisted extraction. 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 Tyrrhenian coast are indicated with the abbreviations R3 (maceration) and R4 (ultrasound-assisted extraction). The chemical composition was analyzed using High Pressure liquid chromatography–Diod-Array detection–Electrospray ionization–Quadrupole–Mass Spectroscopy (HPLC-DAD-ESI-Q-MS). The antioxidant activity was analyzed by 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) 2,2-diphenyl-1-picrylhydrazyl (DPPH), β-carotene bleaching, and Ferric Reducing Antioxidant Power (FRAP) assays. Antioxidant features were also assessed in lipopolysaccharide (LPS)-stimulated RAW-264.7 murine macrophages, evaluating Reactive Oxygen Species (ROS) production; in the same experimental model, the anti-inflammatory activity of the extracts was investigated. Interestingly, all extracts displayed antioxidant and anti-inflammatory properties. They exhibited significative nitrite production inhibitory activity, whith IC50 values ranging from 3.46 to 5.53 µg/mL, without impairing cell viability. The anti-inflammatory activity was also investigated by Western Blotting and immunofluorescence assay, highlighting the R3 and R4 extracts ability to reduce NF-κB translocation, as well as to disrupt the MAPKs signaling pathway. Extracts exhibited both potential anti-proliferative activity on breast cancer cells, inducing apoptosis, without affecting non-tumorigenic cells, and the ability to inhibit MDA-MB-231 cells’ motility. Finally, the rosemary extracts treatment significantly reduced the power of conditioned media, from MCF-7 or MDA-MB-231 cells to induce nitrite production on RAW 264.7 cells, confirming their promising anti-inflammatory activity.
Highlights
Cancer is nowadays one of the biggest health problems in the world
S. rosmarinus aerial parts collected in two different areas of southern Italy were subjected to two extraction procedures namely maceration and ultrasound-assisted extraction by using methanol as solvent
A supercritical fluid rosemary extract dose-dependently induces apoptosis in breast cancer cells from different tumor subtypes including T47D and SK-BR-3 cell lines [63]. On this basis and considering the impact of rosemary extracts tested in this study on MCF-7 and MDA-MB-231 cell viability, we examined if the anti-proliferative activity displayed by them could be due to apoptosis induction
Summary
Cancer is nowadays one of the biggest health problems in the world. In particular, breast cancer is the most common type of carcinoma in women and represents their major cause of death worldwide [1]. Rosemary leaves are extensively used traditionally to treat some diseases This plant has drawn attention due to its biological activities, including antioxidant, antibacterial, hypoglycaemic, anticancer, hepatoprotective, anti-inflammatory, and antithrombotic effects [29,30,31,32,33]. These biological properties have made rosemary a new potential therapeutic agent for the treatment of many diseases. The current study was planned to assess and to compare the chemical composition, the antioxidant power, the anti-inflammatory, and anti-cancer effects of four Italian S. rosmarinus extracts obtained by two extraction procedures and collected in two different localities of southern Italy, when the amount of carnosic acid was negligible
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