本文選題：乳腺癌 + 圓葉葡萄皮渣��； 參考：《中國(guó)農(nóng)業(yè)大學(xué)》2017年博士論文

【摘要】：作為葡萄科中與葡萄屬并列的圓葉葡萄屬,起源于美國(guó)東南部,近幾十年已經(jīng)選育出一系列鮮食和加工品種,并于近年由本實(shí)驗(yàn)室成功引入我國(guó)栽培種植。圓葉葡萄是公認(rèn)的抗性最強(qiáng)的葡萄品種,其果實(shí)除富含多酚物質(zhì)外,還獨(dú)有具較強(qiáng)抗氧化功效的鞣花酸。本文以圓葉葡萄'Noble'以及歐亞種葡萄'赤霞珠'釀酒皮渣為材料,探究酚類提取物各組分抗乳腺癌的功效,并進(jìn)一步了解'Noble'皮渣酚類提取物各組分具體的抗癌效果,最后從不同層面對(duì)其抗癌機(jī)理進(jìn)行探究。兩種葡萄皮渣的多酚在提取后進(jìn)行組分分離,分離后的各組分用于處理MDA-MB-231及MCF-7乳腺癌細(xì)胞。之后利用流式細(xì)胞儀通過(guò)PI染色以及Annexin V-FITC/PI雙染色研究各組分的處理對(duì)細(xì)胞周期及凋亡的影響,并用Western blot對(duì)相關(guān)細(xì)胞因子進(jìn)行檢測(cè)。最后通過(guò)transwell、ELISA等方法研究各組分在癌細(xì)胞轉(zhuǎn)移及預(yù)防(如炎癥因子和ACE的抑制)等其他方面的功效。結(jié)果顯示:1.從兩種不同的葡萄中均分離出三個(gè)組分。對(duì)于'Noble'皮渣提取物,組分1主要為黃酮醇類物質(zhì)和鞣花酸,組分2主要為原花色素和鞣花單寧,而組分3主要為花色素與花色苷類物質(zhì)以及鞣花酸。'赤霞珠'皮渣提取物組分1則以有機(jī)酸和沒(méi)食子酸為主,組分2則是原花色素,組分3則主要為花色素與花色苷類物質(zhì),且各組分均不含鞣花酸類物質(zhì)。2.細(xì)胞活力測(cè)試和半數(shù)抑制濃度結(jié)果分析發(fā)現(xiàn),'Noble'皮渣提取物各組分對(duì)MDA-MB-231的活力抑制強(qiáng)于MCF-7,各組分對(duì)兩株細(xì)胞的抑制能力為組分2≥組分3組分1; '赤霞珠'皮渣提取物的組分2對(duì)MCF-7細(xì)胞的活力抑制更有效,而其余兩個(gè)組分則對(duì)兩株細(xì)胞的抑制能力基本相當(dāng),各組分抑制能力為組分2≈組分1組分3。3.細(xì)胞周期阻滯和細(xì)胞凋亡測(cè)試結(jié)果顯示,對(duì)于MDA-MB-231細(xì)胞,'Noble'皮渣提取物各組分均能通過(guò)下調(diào)cyclin A的表達(dá)使該細(xì)胞阻滯于S期,其中細(xì)胞周期阻滯的能力為組分2≥組分3組分1;各組分可通過(guò)激活caspase 3、caspase 7和caspase 9以誘導(dǎo)細(xì)胞凋亡,其中誘導(dǎo)細(xì)胞凋亡的能力為組分3組分1組分2;對(duì)于MCF-7細(xì)胞,組分2和組分3的處理也可造成S期周期阻滯,其中阻滯能力前者較強(qiáng)。同時(shí)這兩種組分可通過(guò)引起細(xì)胞壞死影響細(xì)胞活力。4.細(xì)胞侵襲實(shí)驗(yàn)結(jié)果顯示,各組分的處理并不能抑制兩株乳腺癌細(xì)胞的侵襲力;抗炎實(shí)驗(yàn)結(jié)果顯示,各組分均能有效減少NO、IL-6以及TNF-α的分泌量,其中組分3的抑制能力最強(qiáng);ACE活力體外抑制實(shí)驗(yàn)結(jié)果顯示,各組分均至少能抑制60 %的ACE活力。葡萄提取物的各組分均能抑制乳腺癌細(xì)胞的活力,并且'Noble'皮渣提取物的各個(gè)組分能通過(guò)細(xì)胞周期阻滯和細(xì)胞凋亡或細(xì)胞壞死對(duì)癌細(xì)胞活力進(jìn)行抑制,并很可能通過(guò)抗炎和抑制ACE活力降低乳腺癌的發(fā)生,具有開(kāi)發(fā)成抗癌試劑或保健品的潛力。
[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，

本文編號(hào)：1937101