Protective Effect Of Aronia Melanocarpa Anthocyanin Extract On Human Retinal Pigment Epithelial Cells

Aronia melanocarpa,also known as wild cherry berry,old berries,contains proanthocyanidins,anthocyanins and flavonoids and other polyphenols.Anthocyanins have strong antioxidant activities,anticancer,antithrombotic,enhancing immunity and promoting the reconstitution of erythropoietin and many other biological activities.In recent years,domestic research on its functionality has increased,but food development and processing is still in its infancy,far less than foreign countries.The optimum extraction technology of Aronia melanocarpa anthocyanins is studied,and the total anthocyanins and their species are identified.In addition,the antioxidant capacity of Aronia melanocarpa anthocyanins is investigated in vitro,and the protective effect of the extracts on the oxidative damage of retinal pigment epithelial cells induced by H₂0₂ is studied.The data is provided for the functional product development of Aronia melanocarpa.The conclusion is as follows:1.Based on single factor experiment,four factors and three levels of response surface test are carried out to optimize the ultrasonic assisted extraction of Aronia melanocarpa anthocyanin.The optimum extraction conditions are determined as follows:ethanol concentration 62%,ultrasonic power 95 W,liquid-solid ratio 19:1?mL/g?and extraction time45 min.Under this conditions,the yield of aronia melanocarpa anthocyanin is 4.248 mg/g with a 1.56%error compared with the actual value.2.Aronia melanocarpa anthocyanins are identified by liquid chromatography-mass spectrometry analysis.5 anthocyanins are identified by chromatogram and mass spectrogram andcomparedwithreferences,includingcyanidin-3,5-dianhexoseglycoside,cyanidin-3-galactoside,cyanidin-3-arabinoside,cyanidin-3-glucoside and cyanidin-3-xyloside.3.Total antioxidant capacity?T-AOC?and 2,2'-azinobis?3-ethylbenzothiazoline-6-sulphonate?radical?ABTS⁺·?and superoxide anion radical(O^2-·)radical scavenging activities of Aronia melanocarpa anthocyanin and VC are comparatively analyzed and the results reveal that the antioxidant activity of Aronia melanocarpa anthocyanin is superior to VC.4.Inverted phase-contrast microscopy,the 3-?4,5-dimethyl-2-thiazolyl?-2,5-diphenyl-2-H-tetrazolium bromide?MTT?assay,2,7-Dichlorodi-hydrofluorescein diacetate?DCFH-DA?fluorescent probes,kit assays and flow cytometry are used for measuring the morphological endothelial features,cell viability,reactive oxygen species?ROS?level,caspase-3 activity and apoptotic cells,respectively.Aronia melanocarpa anthocyanin extract protects RPE cells from damage induced by H₂O₂,as indicated by morphological observation,and the MTT assay shows that the viability of RPE cells is increased from 63.32%to 86.30%.Aronia melanocarpa anthocyanin extract scavenges ROS and reduces caspase-3 activity.Apoptosis in the RPE cells treated by Aronia melanocarpa anthocyanin extract is decreased from 29.11%to 8.85%.Compared with H₂0₂ treatment,it is significantly lower?P<0.01?.The results show that the anthocyanins of Aronia melanocarpa anthocyanin extract can protect the ARPE-19 cells induced by H₂O₂.