Department of Pharmacology, Faculty of Pharmacy, Anadolu University, Eskisehir, Turkey.
J Med Food. 2011 Dec;14(12):1638-46. doi: 10.1089/jmf.2011.0062. Epub 2011 Aug 23.
Pomegranate (Punica granatum L.) is known to possess pharmacological activities, such as antioxidant and anticancer. In this study, we evaluated the antioxidant potency of a methanolic pomegranate fruit peel extract (PPE) and the relation with its antiproliferative and apoptotic effects on MCF-7 human breast cancer cells. Total phenolic content and antioxidant activity of PPE were determined using the Folin-Ciocalteau and the 2,2-diphenyl-l-picrylhydrazyl free radical methods, respectively. Phenolic acids present in the extract were characterized by a reverse-phase high-performance liquid chromatography (HPLC) method. Cell proliferation was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assay. The apoptotic effects were determined by in situ Tdt-mediated dUTP nick end-labeling assay, and Bax/Bcl-2 mRNA expression levels were measured by reverse transcription-polymerase chain reaction. The extraction yield as a percentage of plant material was 37.97% (wt/wt), and total phenolic content was 331.28 mg of gallic acid equivalents/g of extract. According to HPLC analysis, the most abundant phenolic acid detected in the extract was ellagic acid. MCF-7 cell proliferation decreased depending on PPE concentration (25, 50, 100, 200, and 300 μg/mL) and incubation times (24, 48, and 72 hours). After 48 and 72 hours, the apoptotic cell numbers were significantly increased at 100, 200, and 300 μg/mL PPE concentrations. In addition, expression of the pro-apoptotic gene Bax was increased, and that of the anti-apoptotic gene Bcl-2 was decreased after 200 and 300 μg/mL PPE treatment for 48 and 72 hours. Because PPE reduced cell proliferation and induced apoptosis on MCF-7 cancer cells, we believe that PPE has important antioxidant and apoptotic effects.
石榴(Punica granatum L.)具有药理学活性,如抗氧化和抗癌作用。在这项研究中,我们评估了一种石榴皮甲醇提取物(PPE)的抗氧化能力及其对 MCF-7 人乳腺癌细胞的增殖抑制和凋亡作用的关系。使用 Folin-Ciocalteau 和 2,2-二苯基-1-苦基肼自由基法分别测定 PPE 的总酚含量和抗氧化活性。采用反相高效液相色谱(HPLC)法对提取物中的酚酸进行了表征。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物还原法评估细胞增殖。通过原位 Tdt 介导的 dUTP 缺口末端标记法测定凋亡作用,并通过逆转录-聚合酶链反应测量 Bax/Bcl-2 mRNA 表达水平。提取物的提取率为植物材料的 37.97%(重量/重量),总酚含量为 331.28 毫克没食子酸当量/克提取物。根据 HPLC 分析,提取物中检测到的最丰富的酚酸是鞣花酸。MCF-7 细胞增殖随 PPE 浓度(25、50、100、200 和 300 μg/mL)和孵育时间(24、48 和 72 小时)而降低。在 48 和 72 小时后,在 100、200 和 300 μg/mL PPE 浓度下,凋亡细胞数量显著增加。此外,在 200 和 300 μg/mL PPE 处理 48 和 72 小时后,促凋亡基因 Bax 的表达增加,而抗凋亡基因 Bcl-2 的表达减少。由于 PPE 降低了 MCF-7 癌细胞的增殖并诱导了细胞凋亡,因此我们认为 PPE 具有重要的抗氧化和凋亡作用。