Inhibitory Effect Of Black Soybean Extract Anthocyanin On Rat Lens Epithelial Cells

Objective:To discuss about black soybean anthocyanin extractive' effects on rat lens epithelial cells' proliferation,migration and apoptosis.Method:1.Use H202,to induce and make cataract model of rats,and then get the rat lens epithelial cells after oxidation.2.Take microwave assisted method to extract black soybean anthocyanin and choose D101 macroporous resin to purify.3.Choose the selected rat lens epithelial cells,add 1Oug/ml?20ug/ml.30ug/ml?50ug/ml?80ug/ml?lOOug/ml?150 ug/ml concentration's black soybean anthocyanin extractive and take MTT method to observe the proliferation ability of rat lens epithelial cells;Use flow cytometry to detect the apoptosis status of rat lens epithelial cells;Use scratchmethod to detect the migration ability of rat lens epithelial cells.Result:1.In normal tissue culture condition,rat lens epithelial cells can proliferate and grow rapidly.2.Rat lens epithelial cells' proliferation appears different inhibiting effects after adding 1Oug/ml.20ug/ml?30ug/ml?50ug/ml?80 ug/ml?100 ug/ml?150 ug/ml concentration's black soybean anthocyanin extractiveand inhibiting effect is positively correlated with concentration.As time increases,the inhibitory effect becomes more pronounced.IC50 was 98.43ug/ml when anthocyanin extractive dealt with cells after 48 hours,data difference has statistical significance(P<0.05).3.With the increase of anthocyanin's working time,rat lens epithelial cells'apoptosis rate rises gradually,and apoptosis rate has a dependence relationship with time.When black soybean anthocyanin extractive works for 6 h?12h?24h?48 h?72h,the normal group's cell apoptosisratesare5.6+1.3%?13.1 ±8.2%.25.3±4.7%?34.6±9.4%?41.5±13.4%respectively and anthocyanin work group's cell apoptosis rates are 12.6 ± 3.4%?26.8 ± 4.6%?64.1 ± 8.4%?85.3 ± 5.7%?92.1 ± 5.2%respectively,the data of 48 h and 72 h,difference has statistical significance(P<0.05).4.When different concentrations of the drugs scratched cell culture,the migration ability of LECs is different.With the rise of anthocyanin work group's concentration,all of the scratch area migration healing rates are lower than adjacent inferior quality concentration group obviously,the data of 50 ug/ml and 100 ug/ml,difference has statistical significance(P<0.05).When anthocyanin(100 ug/ml)scratched Rat LECs,compared with original scratch area,24 h,48 h and 72 h's cell migration rates increase,48 h and 72 h'scomparative difference between groups has statistical significance(P<0.05).Anthocyanin work group's migration healing rates are lower than normal group,and as time goes by,the differences of cell migration healing rates and normal group are obviously increased,and the differences have statistical significance(P<0.05).Conclusion:1.Black soybean extractive anthocyanin has obvious inhibiting effect on rat lens epithelial cells' proliferation and migration;2.Black soybean extractive anthocyanin has certain induction effect on rat lens epithelial cells'apoptosis;3.Black soybean extractive anthocyanin may have protective effect on PCO.