Regulatory Effect Of Anthocyanins On Oxidative Stress Injury In HepG2 Cells

Oxidative stress is likely to cause functional damage to cells,anthocyanins are safe and non-toxic,and are the most widely available edible antioxidants in nature.In this study,main ingredients of anthocyanins were analyzed and identified by LC-MS from Purified black rice anthocyanin(BRAI),black bean anthocyanin(BSAI),black corn anthocyanin(BCAI),blueberry anthocyanin(BAI),blackberry anthocyanin(BBAI)and red raspberry anthocyanins(RAI).The oxidative stress injury model was induced by oleic acid(OA)for the first time in HepG2 cells,and the oxidative damage of cells was induced by six anthocyanins at a concentration of 50-800 ?g/mL.Total cell viability(T-SOD),antioxidant capacity(T-AOC),lipid peroxidation(MDA),and glutathione peroxidase activity(GSH-PX)were measured.Select BRAI and BAI with the best antioxidant effect to continue the follow-up experiment.Flow cytometry was used to determine the effect of two anthocyanins on apoptosis of HepG2 cells;Detection of mRNA expression levels of GSTP1,HO-1,PPAR?,Keapl,NF-?B and PI3K genes by RT-PCR.Analysis of the expression of related genes in the glutathione signaling pathway.The results are as follows:The main components of the six anthocyanins are:black rice anthocyanins are consist of cyanidin-3-glucoside and peony-3-glucoside;black soybean anthocyanin is consist of cyanidin-3-rutinoside;black corn anthocyanin is consist of cyanidin-3,5-diosgenin-diglucoside;blueberry anthocyanins are consist of cyanidin-3-glucoside and peony-3-glucoside;blackberry anthocyanins are consist of cyanidin-3-glucoside and peony-3-glucoside;red raspberry anthocyanins are consist of cyanidin-3-glucoside,Acridine-3-glucose,saponin-3-glucoside and peony-3-glucoside.HepG2 oxidative damage can be effectively prevented by using six anthocyanins of 0-800 ?g/mL,and T-SOD,T-AOC and GSH-PX tend to decrease first and then increase,MDA showed a trend of increasing first and then decreasing;With the addition of anthocyanin concentration,the intracellular ROS content gradually decreased.When the anthocyanin concentration was 400?g/mL,the above indicators changed most significantly,and BRAI and BAI had the strongest effect.The results of apoptosis experiments showed that with the addition of OA,the number of viable cells decreased to 74.39%,and the ratio of early and late apoptotic cells gradually increased,and the number of dead cells reached 12.55%.With the addition of BRAI and BAI,the number of viable cells decreased to 62.53%and 64.47%,and the rate of apoptotic cells in early and late stages also increased,but the number of dead cells was controlled,10.48%and 9.9%,respectively.It indicates that anthocyanins have a good resistance to oxidative damaged cells.BRAI and BAI significantly up-regulated the relative expression of GSTP1,HO-1 and PPARy genes,and down-regulated the relative expression levels of Keapl,NF-?B and PI3K genes.It indicated that anthocyanins may alleviate oxidative stress damage by regulating glutathione signaling pathway.