DataSheet1_A Polyphenol-Rich Extract of Olive Mill Wastewater Enhances Cancer Chemotherapy Effects, While Mitigating Cardiac Toxicity.DOCX

Cardiovascular toxicity remains one of the most adverse side effects in cancer patients receiving chemotherapy. Extra-virgin olive oil (EVOO) is rich in cancer preventive polyphenols endowed with anti-inflammatory, anti-oxidant activities which could exert protective effects on heart cells. One very interesting derivative of EVOO preparation is represented by purified extracts from olive mill waste waters (OMWW) rich in polyphenols. Here, we have investigated the anti-cancer activity of a OMWW preparation, named A009, when combined with chemotherapeutics, as well as its potential cardioprotective activities. Mice bearing prostate cancer (PCa) xenografts were treated with cisplatin, alone or in combination with A009. In an in vivo model, we found synergisms of A009 and cisplatin in reduction of prostate cancer tumor weight. Hearts of mice were analyzed, and the mitochondria were studied by transmission electron microscopy. The hearts of mice co-treated with A009 extracts along with cisplatin had reduced mitochondria damage compared to the those treated with chemotherapy alone, indicating a cardioprotective role. To confirm the in vivo results, tumor cell lines and rat cardiomyocytes were treated with cisplatin in vitro, with and without A009. Another frequently used chemotherapeutic agent 5-fluorouracil (5-FU), was also tested in this assay, observing a similar effect. In vitro, the combination of A009 with cisplatin or 5-FU was effective in decreasing prostate and colon cancer cell growth, while it did not further reduce growth of rat cardiomyocytes also treated with cisplatin or 5-FU. A009 cardioprotective effects towards side effects caused by 5-FU chemotherapy were further investigated, using cardiomyocytes freshly isolated from mice pups. A009 mitigated toxicity of 5-FU on primary cultures of mouse cardiomyocytes. Our study demonstrates that the polyphenol rich purified A009 extracts enhance the effect of chemotherapy in vitro and in vivo, but mitigates chemotherpy adverse effects on heart and on isolated cardiomyocytes. Olive mill waste water extracts could therefore represent a potential candidate for cardiovascular prevention in patients undergoing cancer chemotherapy.

心血管毒性仍是接受化疗的癌症患者最严重的不良反应之一。初榨橄榄油（Extra-virgin olive oil, EVOO）富含具备抗炎、抗氧化活性的防癌多酚，这类物质可对心肌细胞发挥保护作用。橄榄磨坊废水（olive mill waste waters, OMWW）来源的纯化多酚提取物，是初榨橄榄油制剂中极具研究价值的一类衍生物。本研究针对一款命名为A009的OMWW制剂，探究其与化疗药物联用的抗癌活性，以及潜在的心脏保护作用。构建前列腺癌（prostate cancer, PCa）异种移植模型的小鼠，分别接受顺铂单药治疗或顺铂联合A009治疗。在体内实验中，我们发现A009与顺铂联用可协同降低前列腺癌异种移植瘤的重量。研究人员对小鼠心脏进行了分析，并通过透射电子显微镜观察线粒体形态。与单纯化疗组相比，接受A009提取物联合顺铂治疗的小鼠心脏线粒体损伤程度更低，这表明A009具备心脏保护作用。为验证体内实验结果，本研究在体外分别采用顺铂单药及顺铂联合A009处理肿瘤细胞系与大鼠心肌细胞。另一种临床常用化疗药物5-氟尿嘧啶（5-fluorouracil, 5-FU）也在该实验中得到测试，且取得了相似的实验结果。体外实验表明，A009与顺铂或5-FU联用可有效抑制前列腺癌及结肠癌细胞的增殖，但不会进一步抑制经顺铂或5-FU处理的大鼠心肌细胞的生长。为进一步探究A009对5-FU化疗所致不良反应的心脏保护作用，本研究采用了从新生小鼠体内分离的原代心肌细胞进行实验。A009可缓解5-FU对小鼠原代心肌细胞培养物的毒性作用。本研究证实，富含多酚的纯化A009提取物可在体内外增强化疗药物的疗效，同时减轻化疗对心脏及分离心肌细胞的不良反应。因此，橄榄磨坊废水提取物有望成为癌症化疗患者心血管保护的潜在候选制剂。