YABAN MERSİNİ İÇERİSİNDE BULUNAN HİDROKSİSİNNAMİK ASİDLERİN KANSER HÜCRE KÜLTÜRÜNDE ANTİOKSİDAN AKTİVİTESİ VE APOPİTOSİZ İNDUKSİYONU ÜZERİNE ETKİSİ
Project no. 170 · Status: Kapanmış · 2018
Summary
Recently, wild blueberry is very popular in our country. Blueberries are an aromatic plant that is a promising source of antioxidant compounds with medicinal properties due to their hydroxycinnamic acids. Blueberry fruits that are firming but sweet and edible include flavnoids and Hydroxycinnamic acids (Caffeic, p-Kumaric and Ferulic acid) with strong antioxidant properties. Phenolic compounds found in such foods can act as antioxidants in many potential ways, such as free radical scavenging, oxygen radical absorbance, and chelating of metal ions. Vitamin C, abundant in blueberries, retains free radicals, protecting the DNA of the cell and preventing cancer formation. Antibiotics such as antacids, ellagic acid and urolithin in blueberries also have a protective effect against cancer by reducing the free radical damage of DNA. In this study, wild bleuberry tea obtained by different infusion and boiling methods using the fruits and leaves of naturally grown blueberry plant in our country will be evaluated for the presence of hydroxycinnamic acids, antioxidant activities and apoptosis. Blueberry tea; leaves, raw fruit, dried and chopped raw fruit, fruit seeds and seedless raw fruit in different infusions and boiling: 1 min, 3 min, 5 min, 7 min, 10 min. Phenolic levels will be determined by LC MS / MS technique. The antioxidants and activities of all products in the in vitro HCT-116 colon cancer cell line will be analyzed by spectrophotometric methods. MDA and TEAC were evaluated for antioxidant activity. Cytotoxicity and viability tests were performed by adding WST-8 (Water Soluble Tetrazolium Salt-8) solution. Apoptosis was assessed using the TRAIL and Apaf-1 ELISA Kit methods, which are necessary receptors for the activation of the caspases of the intersecting and extrinsic pathways. As a result, it is the first study evaluating the antioxidant potential of HCT-116 colon cancer cells in hydrocineramic acids in blueberry tea extracts. When the active substances according to the TEAC trolox equivalent were compared, the highest results were calculated as 13.5 mM of blueberry tea extract and the obtained results were evaluated as 5 min boiling suggesting that blueberry tea is a rich source of antioxidants. When we evaluate hydroxysinnamic acids in combination with ferulic, cafeic and p-coumaric acid, which is a hydrocinnamic acid, and combinations thereof, and in vitro studies of cell culture in vivo by wild blueberry alone, p-coumaric acid is the most effective hydrocycinnamic acid with an IC50 concentration of 39.9 µg/mL. It is observed that in double and triple hydrocinnamic acid concentrations, the cytotoxicity of a mixture of IC50 doses at a ratio of 1: 1 of caffeic acid and ferulic acid is 2.6 times higher than that of the cytotoxicity that half of the determined IC50 doses do. The triple combination is still 4.5 times more cytotoxic than the cytotoxic one-half of the IC50 doses of each hydroxycinnamic acid in the ratio 1: 1, making half of the IC50 doses determined to be successful in one single pass. When hydroxycinnamic acid in wild bluberry tea is prepared by 5 min boiling method, it is observed that blueberry tea is more effective than single and combined hydroxycinnamic acid. The highest value obtained by comparing the combinations of these hydrocinnamic acids and the blueberry tea extract, especially caffeic, p-Kumaric, ferulic acid and evaluated and compared of TRAIL, Apaf-1, Cytochrome-C,Caspase 3,Caspase 8, Caspase 9 ELISA kit results.