Brindisi Matteo, Bouzidi Chouaha, Frattaruolo Luca, Loizzo Monica R, Tundis Rosa, Dugay Annabelle, Deguin Brigitte, Cappello Anna Rita, Cappello Maria Stella
Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, Via Pietro Bucci, 87036 Arcavacata di Rende (CS), Italy.
Faculté de Pharmacie de Paris, Université de Paris, U.M.R. n 8038, -CiTCoM (CNRS, Université de Paris), 75006 Paris, France.
Antioxidants (Basel). 2020 Sep 3;9(9):826. doi: 10.3390/antiox9090826.
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 Spenn. in two different sites of Southern Italy obtained by maceration or ultrasound-assisted extraction. Extracts of collected on the Ionian coast are indicated with the abbreviations R1 (maceration) and R2 (ultrasound-assisted extraction). Extracts of 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 IC 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.
在本研究中，我们评估并比较了通过浸渍或超声辅助提取从意大利南部两个不同地点获得的Spenn.的四种甲醇提取物（R1 - R4）的化学成分、抗氧化、抗炎和抗增殖作用。在爱奥尼亚海岸采集的提取物用缩写R1（浸渍）和R2（超声辅助提取）表示。在第勒尼安海岸采集的提取物用缩写R3（浸渍）和R4（超声辅助提取）表示。使用高压液相色谱 - 二极管阵列检测 - 电喷雾电离 - 四极杆 - 质谱联用仪（HPLC - DAD - ESI - Q - MS）分析化学成分。通过2,2'-联氮 - 双（3 - 乙基苯并噻唑啉 - 6 - 磺酸）（ABTS）、2,2 - 二苯基 - 1 - 苦基肼（DPPH）、β - 胡萝卜素漂白和铁还原抗氧化能力（FRAP）测定分析抗氧化活性。还在脂多糖（LPS）刺激的RAW - 264.7小鼠巨噬细胞中评估抗氧化特性，评估活性氧（ROS）的产生；在相同的实验模型中，研究提取物的抗炎活性。有趣的是，所有提取物都表现出抗氧化和抗炎特性。它们表现出显著的亚硝酸盐产生抑制活性，IC值范围为3.46至5.53μg/mL，且不损害细胞活力。还通过蛋白质印迹和免疫荧光测定研究抗炎活性，突出了R3和R4提取物降低NF - κB易位以及破坏丝裂原活化蛋白激酶（MAPKs）信号通路的能力。提取物对乳腺癌细胞表现出潜在的抗增殖活性，诱导细胞凋亡，而不影响非致瘤细胞，并且具有抑制MDA - MB - 231细胞运动的能力。最后，迷迭香提取物处理显著降低了MCF - 7或MDA - MB - 231细胞条件培养基对RAW 264.7细胞诱导亚硝酸盐产生的能力，证实了它们有前景的抗炎活性。
