Studies On Preparation Process Of Isostrictiniin From Nymphaea Candida Presl And Its Hepatoprotective Activity

Objective:To study on hepatoprotective activity and preparation process of isostrictiniin from Nymphaea candida.Methods:The extraction process and macroporous resin adsorption purification process of isostrictiniin from N candida were optimised using single factor experiment combined with orthogonal experiment design.Hepatoprotective activity of isostrictiniin was evaluated through carbon tetrachloride(CCl4)induced chemical liver injury and Concanavalin A(Con A)induced immunological liver injury in mice,and its antioxidant effects were investigated by reducing power,DPPH radical scavenging experiments and total antioxidant capacity determination.Result:The extract conditions were optimized as follows:extracting time was 90 min,extraction times was twice,solvent concentration was 50%ethanol and solid-liquid ratio was 1:50.The average extraction rate and yield of isostrictiniin were 44.46%and 0.95%respectively.The H1020 macroporous adsorption resin was used to purify isostrictiniin with the optimum conditions as follows:sample concentration at 0.354 mg/mL,sample velocity at 2.0 mL/min,and desorption rate at 2.0 mL/min.The transfer rate and the purity of isostrictiniin was 10.33%and 92.42%were determined and calculated by HPLC respectively.This optimized purification process was stable and feasible.Isostrictiniin had a strong scavenging effect on DPPH radical with a concentration-response relationship,IC50 value of isostrictiniin and Vit C were 8.95 and 6.33g/mL respectively.Compared with CCl4 induced acute liver injury in mice,serum ALT,AST levels and liver spleen index were decreased(P<0.05)and the contents of liver homogenate MDA and NO were decreased,and the activities of liver homogenate SOD and GSH were increased by isostrictiniin(50,100 mg/kg)(P<0.05).Histopathological examination results showed that isostrictiniin(50,100 mg/kg)could reduce the CCl4 induced liver pathological damage in mice.Compared with Concanavalin A induced acute liver injury in mice,the serum ALT,AST,TNF-α,IL-1β,IL-6 as well as liver and spleen index were significantly reduced by isostrictiniin(50,100 mg/kg)(P<0.05);The contents of MDA,NO,IL-10,IL-18 and activities of SOD,GSH,IL-2,and IL-4 were increased in homogenate liver tissue by isostrictiniin(P<0.05).The histopathological changes of liver were improved obviously also.Conclusion:The preparation process of isostrictiniin is simple and easy to produce,and this compound has better liver protection activity.These results can provide data basis for the development and utilization of N.candida and this compound.