New triterpenoid saponin compound with liver protection effect
A technology of triterpene saponins and compounds, which is applied in the field of medicine, can solve problems such as inconvenient use, high price, and serious side effects, and achieve the effect of enhancing SOD activity and protecting the liver
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Embodiment 1 3
[0025] The preparation of embodiment 1 triterpene saponin compound
[0026] Weigh 80 kg of licorice slices, add 10 times the amount of 95% ethanol to heat and reflux to extract twice, each time for 2 hours, filter, add 10 times the amount of 50% ethanol to the dregs and repeat the extraction twice, filter, combine the extracts and decompress Concentrate until there is no alcohol smell; add appropriate amount of water to the extract to form a suspension, extract the suspension with petroleum ether and ethyl acetate; prepare the extraction residue with an aqueous solution to a certain concentration, elute through D101 macroporous resin, and first use 5 column volumes were eluted with pure water with a pH of 1-2, and then 5 column volumes were eluted with 10% ethanol with a pH of 1-2, and finally eluted with 50% ethanol with a pH of 1-2, and 50% ethanol The total licorice saponin extract was obtained by dehydration and drying (yield 4.2kg).
[0027] Total saponin part by C 18 M...
Embodiment 2
[0030] Example 2 Experimental Research on Anti-liver Damage of New Triterpenoid Saponin Compounds
[0031] 1. Experimental materials and drugs
[0032] 1. Drugs and reagents
[0033] AST, ALT, MDA, SOD kits and Coomassie brilliant blue protein kits were purchased from Nanjing Jiancheng Bioengineering Institute; carbon tetrachloride (CCl 4 , analytically pure), used to prepare 0.1% peanut oil solution with peanut oil; galactosaminoside (D-GalN, Sigma company), dissolved in distilled water before use and diluted to 70mg / mL solution; bifendate tablets (Jiangsu Peng Harrier Pharmaceutical Co., Ltd.), grind it into a fine powder and add saline to make a suspension before use.
[0034] 2. Experimental animals
[0035] Clean-grade male ICR mice, weighing 18-22 g, were provided by the Experimental Animal Center of Zhejiang Province, certificate number SCXK (Zhejiang) 2008-0033.
[0036] 3. Experimental equipment
[0037] ELISA (Bio-Tek, USA); UV-2000 ultraviolet spectrophotometer...
Embodiment 3
[0069] Example 3 Cytotoxicity test
[0070] Human hepatocyte L02 cell line was cultured in DMEM medium containing 10% (volume fraction) inactivated standard fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin at 37°C, 5% CO 2, cultivated in an incubator with saturated humidity, and passaged once every 2-3 days. Cells in logarithmic growth phase were used in the experiments. Cells in the logarithmic growth phase were taken, and after trypsinization, the cell suspension was prepared with DMEM medium containing 10% fetal bovine serum, and the cell concentration was about 1×10 5 1 / ml, inoculated in a 96-well culture plate, 180 μL per well; take the saponin compound uralsaponinI prepared in Example 1 of the present invention, respectively set 2 μM, 5 μM, 10 μM, 20 μM, 50 μM, 100 μM 6 concentrations, and then add them to each well 20 μL dimethyl sulfoxide, set 4 duplicate wells for each group, set at 37°C, 5% CO 2 After culturing in the incubator for 72 hours, add...
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