| Anthocyanins has many kinds of physiological activity function, such as elimination free radical, resist caducity and variation, resisting inflammation, improvement eyesight, prevention and treatment of cardiovascular diseases, cancer prevention, enhancement immunity. Study the physiological activity function of anthocyanins from Lonicera edulis (Lonicera edulis Turcz.) has very important practical significance.This paper study the extraction conditions, the oxidation resistance and antitumor function mechanism of anthocyanins after isolation and purification from Lonicera edulis.The optimum condition of extract anthocyanins from Lonicera edulis was discussed by the orthogonal experiment through solvent digesting. The UV spectrophotometer was used to determine the content of anthocyanin of extraction, while H2O2/Fe system and DPPH assay method was adopted to determine the antioxidative capacity of extraction. The results indicated that solvent showed the remarkable effect on antioxidant of anthocyanin from Lonicera edulis, while the ratio of material to liquid, extraction temperature and time showed minor effects. Optimum extraction processing parameter was determined as follow:ethanol concentration was 70%, ratio of material to liquid was 1:15(W:V), temperature was 60℃, time was 90min, pH value was 2, extraction times was third.Anthocyanins from Lonicera edulis of different concentrations was administered to human colon HT29 Cells, lier cancer HepG2 cells and U-2OS cells of human osteosarcoma for 24h,48h and 72h, respectively. The proliferation of cells was measured by MTT assay. The results showed that anthocyanins from Lonicera edulis can inhibit three types of tumor cells showed dose-dependent and time-dependent. The inhibition rate increased significantly with the increased of the concentration of anthocyanins and treatment time.HT29 cells were treated with anthocyanins in different concentration for 48 h, flow cytometry with annexin V FITC/PI double staining was used to detect the apoptosis of the cells. Results:The number of apoptotic cells increases with the increase of anthocyanin concentration. The apoptosis rate of the groups (0.25,0.5,1,2 mg/mL) was (10.11±1.84)%, (28.87±2.32)%, (54.74±4.69)%(73.56±8.34)%. Compared with the control group, the apoptosis rate of the drug groups were significantly (P<0.05).Many typical apoptotic morphology were observed, such as:the number of cells decreased, the volume shrunk, many spherules heaved out cells surface, apoptotic bodies appear, connections between cells gradually fracture, the vacuole inside plasm obviously increased, cytoplasm condense, dense nuclear matter was patchy or crescent-shaped on boundary of the nuclear membrane, nuclear shape was irregular. A typical DNA degradation associated with apoptosis was determined by DNA agarose electrophoresis. Single-celled gel electrophoresis showed cells visible delay phenomenon. Mitochondrial membrane potential was decreased and significantly lower than normal cellular level; Caspase-3 activity was increased with the increase of anthocyanins concentration and the difference was significantly compared to control cells(P<0.05).The results suggest that anthocyanins from Lonicera edulis inhibit cancer cells proliferation maybe was related with apoptosis through the mitochondrial apoptotic signaling pathway.
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