Preparation And Preliminary Study On Anti-oxidant Activity Of Rutin-loaded Liposome

Objective To prepare rutin-loaded liposome with stable quality and evaluate thedrug content, encapsulation efficiency, particle size and stability. A preliminary studyon anti-oxidant activity by comparing the protective effect of rutin-loaded liposomeand rutin aqueous solution on the human umbilical vein endothelial cells (HUVECs)injury induced by hydrogen peroxide (H2O2).Methods The rutin-loaded liposome were prepared by film dispersion method.Evaluating encapsulation efficiency for main index, choose the best prepared craftthrough the single factor experiment and orthogonal design. The content of rutin andencapsulation efficiency was determined by RP-HPLC. Following after the establi-shment of a model of the H2O2-induced HUVECs injury, the cellular viability wasdetected by MTT assay. Contents of nitric oxide synthase (NOS), malondialdehyde(MDA) and lactate dehydrogenase (LDH) in supernatant were measured by the co-mmercial kits.Results The optimum preparation technology of rutin-loaded liposome were deci-ded, film dispersion was used for the preparation of rutin-loaded liposome consistingof egg phosphatedylcholine (EPC)﹕cholesterol (Ch)(5﹕1),drug﹕EPC (1﹕40), or-ganic phase with absolute alcohol and aqueous phase with phosophate buffered saline(PBS pH=6.8) in the ratios of (1﹕1.5) respectively. Rutin-loaded liposome showedintact and spherieal in shape with buffy. Its average size was147.2nm, zeta potentialwas-20±5mv,average drug loading was56.95μg·mL-1and encapsulation efficiencywas66.5%. Stability investigation showed rutin-loaded liposome was stable duringthe storage of6months under4℃. Rutin was well-separated with peaks of the adjuv-ants through RP-HPLC and had a good linear relation in the range from4to80μg·mL-1(r=0.9998, n=6). The average recovery was99.4%. The results of anti-oxidant activity compared with the same concentration rutin aqueous solution showed thatrutin-loaded liposome significantly enhanced protection function of HUVECs fromH2O2-induced injury, increased the contents of NOS and decreased the contents ofMDA and LDH in supernatant.Conclusion Optimized the film dispersion technology was suitable for preparingrutin-loaded liposome with stable quality. RP-HPLC was fit for determining drugcontent and encapsulation efficiency of rutin-loaded liposome. Rutin-loaded liposomehad better antioxidant ability than the same concentration rutin aqueous solution.