RESUMO
Objective:To optimize the extraction process for the total alkaloids of Longzuan Tongbi Fang by central composite design and response surface method.Methods:The independent variables were the ethanol contention,solvent consumption and extraction time,and the dependent variable was the weighted overall desirability of the content of total alkaloids and kukoline.The extraction process for the total alkaloids of Longzuan Tongbi Fang was optimized by central composite design and response surface method.Results:The optimum extraction conditions were as follows:9-fold amount of solvent was used,refluxed for 130 min,extracted twice,and the ethanol concentration was 75%.Conclusion:The central composite design and response surface method used for the extraction optimization for the total alkaloids of Longzuan Tongbi Fang is very simple and highly predictive.
RESUMO
Objective: To observe the protective effects of the alcohol extract, ethyl acetate extract and 1-butanol extract of Euonymus hederaceuson on H2O2-induced oxidative damage of vascular endothelial cells ECV-304.Methods: ECV-304 cells were cultured in vitro and H2O2-induced oxidative damage in ECV-304 cells was used as the model.After the treatment with ethanol extract, ethyl acetate extract or 1-butanol extract of Euonymus hederaceus (the final concentration of 400, 200, 100, 50 and 25 μg·ml-1) for 24h, the cell viability was detected by MTT assay.Hoechst staining was used to observe the cell apoptosis, and then the supernatant was collected, and the contents of SOD, NOS and NO were used to determine the effects of the extracts on the oxidative stress of vascular endothelial cells.Results: The ethanol extract, ethyl acetate extract and 1-butanol extract of Euonymus hederaceus within the concentration range of 25-400 μg·ml-1 showed no cytotoxicity on ECV-304 cells, and the ethyl acetate extract at the concentration of 400μg·ml-1 had certain proliferation effect on the cells (P<0.05).The ethyl acetate extract at the concentration of 50-400 μg·ml-1 and the 1-butanol extract at the concentration of 100-400 μg·ml-1of Euonymus hederaceus could increase the survival rate of ECV-304 cells with oxidative injury significantly, and the OD value showed significant difference when compared with that of the model group (P<0.05 or P<0.01).The ethanol extract, ethyl acetate extract and 1-butanol extract of Euonymus hederaceus could significantly improve the SOD and NO levels in ECV-304 cells with oxidative damage (P<0.05 or P<0.01), and had no notable influence on NOS level.The ethanol extract, ethyl acetate extract and 1-butanol extract of Euonymus hederaceus could inhibit the cell apoptosis with oxidative damage and improve the nuclear morphology, and the apoptosis exhibited significant difference when compared with that of the model group (P<0.01).Conclusion: The extracts of Euonymus hederaceus have protective effects on vascular endothelial cells with oxidative damage induced by H2O2.
