Oliveira Tatiane, Figueiredo Camila A, Brito Carlos, Stavroullakis Alexander, Prakki Anuradha, Da Silva Velozo Eudes, Nogueira-Filho Getulio
Department of Bioregulation, Institute of Health Sciences, Federal University of Bahia, 41110-100 Salvador, BA, Brazil ; Department of Biological and Diagnostic Sciences-Preventive Dentistry, University of Toronto, Toronto, ON, Canada M5G 1G6.
Department of Bioregulation, Institute of Health Sciences, Federal University of Bahia, 41110-100 Salvador, BA, Brazil.
Int J Cell Biol. 2014;2014:535789. doi: 10.1155/2014/535789. Epub 2014 Aug 3.
Allium cepa L. is known to possess numerous pharmacological properties. Our aim was to examine the in vitro effects of Allium cepa L. extract (AcE) on Porphyromonas gingivalis LPS and Escherichia coli LPS-stimulated osteoclast precursor cells to determine cell viability to other future cell-based assays. Osteoclast precursor cells (RAW 264.7) were stimulated by Pg LPS (1 μg/mL) and E. coli LPS (1 μg/mL) in the presence or absence of different concentrations of AcE (10-1000 μg/mL) for 5 days at 37°C/5% CO2. Resazurin reduction and total protein content assays were used to detect cell viability. AcE did not affect cell viability. Resazurin reduction assay showed that AcE, at up to 1000 μg/mL, did not significantly affect cell viability and cellular protein levels. Additionally a caspase 3/7 luminescence assay was used to disclose apoptosis and there was no difference in apoptotic activity between tested groups and control group. Fluorescence images stained by DAPI showed no alteration on the morphology and cell counts of LPS-stimulated osteoclast precursor cells with the use of AcE in all tested concentrations when compared to control. These findings suggest that Allium cepa L. extract could be used for in vitro studies on Porphyromonas gingivalis LPS and Escherichia coli LPS-stimulated osteoclast precursor cells.
已知洋葱(Allium cepa L.)具有多种药理特性。我们的目的是研究洋葱提取物(AcE)对牙龈卟啉单胞菌脂多糖(Pg LPS)和大肠杆菌脂多糖(E. coli LPS)刺激的破骨细胞前体细胞的体外作用,以确定细胞活力,为未来其他基于细胞的试验做准备。在37°C/5% CO₂条件下,将破骨细胞前体细胞(RAW 264.7)在存在或不存在不同浓度(10 - 1000μg/mL)AcE的情况下,用Pg LPS(1μg/mL)和E. coli LPS(1μg/mL)刺激5天。采用刃天青还原法和总蛋白含量测定法检测细胞活力。AcE不影响细胞活力。刃天青还原试验表明,高达1000μg/mL的AcE对细胞活力和细胞蛋白水平无显著影响。此外,使用半胱天冬酶3/7发光试验来揭示细胞凋亡情况,测试组和对照组之间的凋亡活性没有差异。与对照组相比,用DAPI染色的荧光图像显示,在所有测试浓度下,使用AcE处理的LPS刺激的破骨细胞前体细胞的形态和细胞计数均无变化。这些发现表明,洋葱提取物可用于对牙龈卟啉单胞菌脂多糖和大肠杆菌脂多糖刺激的破骨细胞前体细胞的体外研究。