Optimization Of Extration Technology Of Total Flavonids From Rubus Reflexus Ker Var. Reflexus And Its Hepatoprotective Effect

The mechanism of liver injury is one of the hot spot of medical research,rubus plant Rubus reflexus Ker var.reflexus in the treatment of liver disease has good folk medicine foundation.In this study,we investigated the extraction and purification technology of Rubus reflexus Ker var.reflexus total flavonoids and based on the result,we discussed the effect and mechanism of Rubus reflexus Ker var.reflexus total flavonoids on hepatoprotective.First,through the single factor experiment,we studied temperature ethanol concentration,extracting time and material to liquid ratio about extraction technology of Ker var.reflexus total flavonoids.Accroding to the result of the single factor experiment,we determined the best extraction technology by orthogonal experiment.Second,we studied on temperature,sample concentration,sample volume,pH value,sample flow velocity,ethanol volume about purification technology of Rubus reflexus Ker var.reflexus total flavonoids use macroporous adsorption resin,and we determined the best technology of purification.We established the model of acute liver injury induced by CCl4with the bifendate as positive control group,Rubus reflexus Ker var.reflexus total flavonoids as treatment group.Study the influence on liver injury in mice by Rubus reflexus Ker var.reflexus total flavonoids groups compared with the model group through investigating serum ALT,AST,TC,LDL-C level and liver tissue MDA,SOD,GSH index.The results showed that the optimal conditions were as follows:material to liquid ratio is 1:25,60%ethanol concentration,temperature is 70℃,extract 2.5 hours two times.Through the best extraction technology,the yield of Rubus reflexus Ker var.reflexus total flavonoids can be up to 6.78%.The best purification technology is:50%ethanol as elution agent,material to liquid ratio is 1:25,concentration of sample solution for 6.83mg/ml,sample pH for 4,flow velocity of sample was 33BV/h,sample volume is 7BV,elution volume is 3BV water and 6BV 50%ethanol.Through the best purification technology,the yield of Rubus reflexus Ker var.reflexus total flavonoids can be up to 87%.Besides,we aimed to investigate protective effect of total flavonoids from var.reflexes against carbon tetrachloride(CCl4 induced hepatotoxity in mice::ALT level in middle(100 mg/kg)and high concentration(200 mg/kg)flavonoids groups enable decreased significantly(P<0.01);AST level in high concentration flavonoids group make decreased significantly(P<0.01);LDL-C level in low(50 mg/kg)and middle concentration flavonoids groups make decreased significantly(P<0.01);MDA level in high concentration flavonoids group make decreased(P<0.05);SOD level in middle and high concentration flavonoids groups make increased significantly(P<0.01);GSH level in middle and high concentrations flavonoids groups increased significantly(P<0.01).The results showed that total flavonoids from Rubus reflexus Ker var.reflexus has the hepatoprotective actions.The inhibition of CCl4-nduced acute liver injury and inflammation is due at least in part to its antioxidant effects,hepatocellular function regulation and hepatic metabolism modification effects.