Study on Antioxidant and Anticancer Activity of Polyphenols in Dry Red Wines from Eastern Foot of Helan Mountain in Ningxia

Author:Sun Hai Yan

Supervisor:wang hua


Degree Year:2019





Phenols are one of the major secondary metabolites of grapes.There are lots of studies demonstrating that the contents and types of polyphenol not only have the decisive effect on the sensory quality of wines,but also have important impact on the antioxidant ability of wines.Most of the studies focused on the antioxidant activity in vitro of polyphenols in wine and the methods which were developed to detect the antioxidant activity of polyphnols are limited to chemical techques.However,there are little comprehensive and systematic studys on oxidant activity vitro and viviof polyphenols in wines with different grapes viarieties and/or from different years.Therefore,in the present study,we take dry red wines from eastern foot of Helan Mountain as test material to determine the contents of differents kins of phenolic compounds and antioxidant capacity in vitro,analyzing the correlation between the polyphenols and antioxidant ability.Meantime,the antioxidant activities of cancer cells were studied to further reveal the relationship between polyphenols in wines and the antioxidant as well as the antiproliferative activities of cancer cells.Finally,the effects of polyphenols in grape wine on angiogenesis were studied by living animal experiments with ebrafish as materials.The results provided new idea for anticancer researches.The main results of this project are as follows:(1)The contents of phenolic compounds in dry red wines from eastern foot of Helan moutain were determined.The results showed that there was a great difference in the content of phenolic compounds,which was closely related to the grape varieties and years of wines.Cabernet Sauvignon wine in 2013 year has the highest contents of total pheol,total flavonoid,anthocyanin monomer among the wines made with Cabernet Sauvignon but from four different years and the contents are 464.21 mg/L,519.60 mg/Land 10.28 mg/L for total pheol,total flavonoid,anthocyanin monomer.The gallic acid,catechin and resveratrol contents are highest in 17 kinds of monomer phenolic.Investigations on contents of polyphenol compounds in wines with different varieties of grapes were conducted.Results indicated that total flavanol content,tannin content,anthocyanin content,total flavonoid content,anthocyanin monomer and monomer phenolic content in Shiraz Wine were the highest and there are 610.69 mg/L、18.87 mg/L、58.65 mg/L、121.61 mg/L、533.55 mg/L and 86.53mg/L,respectively.(2)Seven kinds of methods were used to study the in vitro antioxidant capacity of wines.Meantimes,the corelations between the contents of polyphenol in wnes and the antioxidant activity were analysized.The results indicated that Fe2+chelating ability in Cabernet Sauvignon wines from different years are mainly affected by the total anthocyanin content and tannin content;anti superoxide anion capacity is mainly determined by the total phenol content.The correlation between phenolic compounds in wines with different varieties and antioxidant activity was investigated.Results indicated that the anti superoxide anion activity was affected by the total phenol content,total flavonoids and flavanols content;The free radical scavenging capacity of DPPH is mainly determined by the total anthocyanins;Fe2+chelating ability relates to total phenol content;Hydroxyl radical scavenging ability were positively affected by the flavanol content.(3)cholesterol and cellular antioxidant model were constructed in this study.Results indicated that wines have great inhibitory activity against the oxidation of human hepatoma cell HepG2.However,the inhibition activities were affected by the years and grape varieties of wines.The Cabernet Sauvignon wines in 2013 year is of highest inhibitory activities against the oxidation of cholesterol and human hepatoma cells HepG2 among the Cabernet Sauvignon wines from four different years.Shiraz Wine holds the highest inhibitory activities against the oxidation of cholesterol and human hepatoma cells HepG2 among the wines with four different grape varieties.Results also suggested that inhibitory efficients are significantly corelated with the content of total phenols,flavanols content and DPPH inhibitory rate.(4)Three common intestinal cancer cells and one normal cell were selected for cell proliferation,migration and apoptosis tests.The mechanisms of cell proliferation and apoptosis were preliminarily explored.The experiment showed that the wine polyphenol extract had inhibitory effect on the proliferation of three colorectal cancer cells,especially the human colorectal cancer cell line HCT116,which IC50 value was 0.17 mg/mL.It was followed by HCT15,HT29 and 293T cells which IC50 values were 0.25 mg/mL,0.27mg/mL,and 0.43 mg/mL,respectively.Meanwhile,wine polyphenols inhibited the migration of tumor cells and promoted cell apoptosis,especially the promotion of HCT116tumor cell apoptosis.Based on clustering heat maps and enrichment maps of genes,the changes in cell proliferation and apoptosis were observed.Compared to normal cell line,the down-regulated gene of T293 cell line and HCT116 with wine polyphenol treatment contributed to major proportion.It indicated that the wine extract can inhibit the proliferation of tumor cells and had anti-disease effects.(5)Effects of polyphenols in wines on zebrafish angiogenesis were studied using zebrafish as an animal model.It was found that the wine polyphenols extracts had no obvious toxicity on zebrafish embryos,and had a good inhibitory effect on zebrafish angiogenesis.Wines with different grape varieties have different effects on angiogenesis inhibition.However,the phenol extract from grape skins had no obvious effect on zebrafish angiogenesis.