| Red raspberry(Rubus idaeus L.)is a Rubus plant of the Rosaceae family.The studies have shown that red raspberry has anti-inflammatory,anti-oxidant,anti-bacterial effects and other activities.With the deepening of the functional study of anthocyanins,it is of great significance for the identification of anthocyanins components of red raspberry and more in-depth of a functional research.Therefore,during this experiment we extracted anthocyanins from red raspberry,further purified and structurally identified,and then used D-galactose-induced aging mice and lung cancer A549 cells in-depth of functional studies to provide some data reference for the future application of red raspberry anthocyanins in food and medicine,mainly from the following four aspects:1.Ultrasound-assisted enzymatic method was used to extract anthocyanins from red raspberry.The optimum extraction process of anthocyanins was obtained by single factor experiment and response surface optimization.The results showed that the enzyme dosage was 5 mg/g,the ratio of material to liquid was 1: 15(g/mL),the enzyme pH was 3.0,the enzyme temperature was 50 ?,the ultrasonic time was 20 min,the ultrasonic power was 450 W and the enzyme time was 60 min.Under these conditions,the anthocyanins content was 127.51 mg/100 g.The combined ultrasound assisted enzymatic method gave a yield of anthocyanins which was 46.38% more than the ultrasonic method and 23.88% more than the enzymatic method respectively.This indicates that the extraction effect of the combined extraction is more advantageous.2.By comparing the adsorption and desorption capacity of macroporous resins,AB-8 was selected from a list of AB-8,D101 and D406.40?g/ml anthocyanins concentration,pH3.0 and at a speed of 2ml/min was added into AB-8 and the anthocyanins was eluted by 90% ethanol.The anthocyanins color of red raspberry purified by AB-8 was 61.3±0.22,the purity of red raspberry anthocyanins was 27.3±0.48%,and the anthocyanins species of red raspberry identified by UPLC-Q/TOF-MS were:paeonidin-3-glucoside,cyanidin-3-glucoside,cyanidin-3-arabinoside and cyanidin-3-rutinoside.3.Through the intervention of red raspberry anthocyanins on aging model mice,the anti-oxidative activity of red raspberry anthocyanins was determined by the index of organs of the mice,the detection of antioxidant enzyme activity in the organs and serum,and the pathological observation of the heart and kidney.The anti-aging effect was assessed.The results showed that the anthocyanins of red raspberry had a certain improvement on the decline of thymus and spleen index caused by aging,which significantly increased the activity of SOD,CAT and GSH-Px in organs and serum of aging mice and decreased the content of MDA,and also significantly increased HYP content of the skin tissue,which played a positive role in improving visceral and skin aging;pathological sections showed cardiomyocytes and kidneys after intervention with red raspberry anthocyanins:The cells structure were clear,the quantity was increased,and the recovery was close to the normal state,indicating that the red raspberry anthocyanins could alleviate the oxidative damage caused by free radicals and inhibit the aging.4.The effects of red raspberry anthocyanins on lung cancer A549 cells were evaluated by MTT assay,cell morphology observation and flow cytometry.The results showed that the inhibition rate of A549 cells treated with red raspberry anthocyanins for 48 hours was 80.39%.The inverted microscope showed that the adhesion ability of lung cancer cells A549 under the action of 100 ?g/mL anthocyanins was decreased,and the cells became round or irregular.The apoptosis rate of the 100 ?g/mL anthocyanins treatment group was 40.53%.It indicated that the red raspberry anthocyanins have the effect of inhibiting the proliferation of lung cancer A549 cells. |
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