葡萄皮渣酚类化合物体外抑制乳腺癌的功效及机理

发布时间：2018-05-26 11:39

  本文选题：乳腺癌 + 圆叶葡萄皮渣　； 参考：《中国农业大学》2017年博士论文

【摘要】：作为葡萄科中与葡萄属并列的圆叶葡萄属,起源于美国东南部,近几十年已经选育出一系列鲜食和加工品种,并于近年由本实验室成功引入我国栽培种植。圆叶葡萄是公认的抗性最强的葡萄品种,其果实除富含多酚物质外,还独有具较强抗氧化功效的鞣花酸。本文以圆叶葡萄'Noble'以及欧亚种葡萄'赤霞珠'酿酒皮渣为材料,探究酚类提取物各组分抗乳腺癌的功效,并进一步了解'Noble'皮渣酚类提取物各组分具体的抗癌效果,最后从不同层面对其抗癌机理进行探究。两种葡萄皮渣的多酚在提取后进行组分分离,分离后的各组分用于处理MDA-MB-231及MCF-7乳腺癌细胞。之后利用流式细胞仪通过PI染色以及Annexin V-FITC/PI双染色研究各组分的处理对细胞周期及凋亡的影响,并用Western blot对相关细胞因子进行检测。最后通过transwell、ELISA等方法研究各组分在癌细胞转移及预防(如炎症因子和ACE的抑制)等其他方面的功效。结果显示:1.从两种不同的葡萄中均分离出三个组分。对于'Noble'皮渣提取物,组分1主要为黄酮醇类物质和鞣花酸,组分2主要为原花色素和鞣花单宁,而组分3主要为花色素与花色苷类物质以及鞣花酸。'赤霞珠'皮渣提取物组分1则以有机酸和没食子酸为主,组分2则是原花色素,组分3则主要为花色素与花色苷类物质,且各组分均不含鞣花酸类物质。2.细胞活力测试和半数抑制浓度结果分析发现,'Noble'皮渣提取物各组分对MDA-MB-231的活力抑制强于MCF-7,各组分对两株细胞的抑制能力为组分2≥组分3组分1; '赤霞珠'皮渣提取物的组分2对MCF-7细胞的活力抑制更有效,而其余两个组分则对两株细胞的抑制能力基本相当,各组分抑制能力为组分2≈组分1组分3。3.细胞周期阻滞和细胞凋亡测试结果显示,对于MDA-MB-231细胞,'Noble'皮渣提取物各组分均能通过下调cyclin A的表达使该细胞阻滞于S期,其中细胞周期阻滞的能力为组分2≥组分3组分1;各组分可通过激活caspase 3、caspase 7和caspase 9以诱导细胞凋亡,其中诱导细胞凋亡的能力为组分3组分1组分2;对于MCF-7细胞,组分2和组分3的处理也可造成S期周期阻滞,其中阻滞能力前者较强。同时这两种组分可通过引起细胞坏死影响细胞活力。4.细胞侵袭实验结果显示,各组分的处理并不能抑制两株乳腺癌细胞的侵袭力;抗炎实验结果显示,各组分均能有效减少NO、IL-6以及TNF-α的分泌量,其中组分3的抑制能力最强;ACE活力体外抑制实验结果显示,各组分均至少能抑制60 %的ACE活力。葡萄提取物的各组分均能抑制乳腺癌细胞的活力,并且'Noble'皮渣提取物的各个组分能通过细胞周期阻滞和细胞凋亡或细胞坏死对癌细胞活力进行抑制,并很可能通过抗炎和抑制ACE活力降低乳腺癌的发生,具有开发成抗癌试剂或保健品的潜力。
[Abstract]:As a juxtaposition of the genus Grape in the family Grape, originated from the southeastern United States, a series of fresh and processed varieties have been selected in recent decades, and have been successfully introduced into China for cultivation by our laboratory in recent years. Round leaf grape is recognized as the most resistant grape variety, its fruit is rich in polyphenols, but also has strong antioxidation effect of tannic acid. In this paper, the effects of phenolic extracts on breast cancer were investigated by using Noble 'and Cabernet Sauvignon' wine dregs of Eurasian grape as materials, and the specific anticancer effects of novolac extracts from Noble 'peel were also studied. Finally, the anti-cancer mechanism is explored from different aspects. Two kinds of grape residue polyphenols were separated after extraction. The separated components were used to treat MDA-MB-231 and MCF-7 breast cancer cells. Flow cytometry and Annexin V-FITC/PI double staining were used to study the effect of each component on cell cycle and apoptosis, and the relative cytokines were detected by Western blot. Finally, transwelling Elisa was used to study the effects of each component on cancer cell metastasis and prevention (such as the inhibition of inflammatory factors and ACE). The result is 1: 1. Three components were isolated from two different grapes. For Noble 'dregs extract, component 1 is mainly flavonol and tannic acid, component 2 is mainly proanthocyanidins and tannins, while component 3 is anthocyanins and anthocyanins, as well as tannic acid. Component 1 of Cabernet Sauvignon 'dregs was mainly organic acid and Gallic acid, component 2 was proanthocyanidins, and component 3 was mainly anthocyanins and anthocyanins. The results of cell viability test and half inhibitory concentration analysis showed that the inhibition of MDA-MB-231 activity of each component of Noble' dregs extract was stronger than that of MCF-7. The inhibitory ability of each component on two cell lines was 2 鈮，

本文编号：1937101