Triterpenoid AntcinK and liver-protecting activity and application thereof
A 25r-antcink, triterpenoid technology, applied to the triterpenoid Antcin K and its hepatoprotective activity and application fields, can solve the problem that the pharmacological activity of Antrodia cinnamomea is not fully studied, the medicinal materials are scarce, and the fingerprint of the extraction method of Antrodia camphorata is not declared. problem, to achieve the effect of protecting the activity of abnormal lipid accumulation and significantly preventing and curing
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Embodiment 1
[0033] Embodiment 1, the separation and purification of triterpenoid Antcin K in Antrodia camphorata
[0034] 1. Experimental materials and methods.
[0035] All chemical reagents mentioned in this method were purchased from Beijing Chemical Plant.
[0036] First take 500 g of dried Antrodia camphorata fruiting body cultured on a plate and pulverize into coarse powder, divide into two batches, and use 6L 95% ethanol to reflux extract each batch for 6 times, each time for 3 hours; combine the extracts and rotary steam under reduced pressure to obtain about 200 g of total extract; After the total extract was suspended in 4 L of water, it was extracted 5 times with 4 L of petroleum ether successively, and the aqueous phase was extracted 5 times with ethyl acetate; the combined extract was evaporated to dryness to obtain about 132 g of the ethyl acetate layer.
[0037] About 130g of the extract from the ethyl acetate layer was separated through a silica gel column (200-300 mesh, Qi...
Embodiment 2
[0038] Embodiment 2, triterpenoid Antcin K in Antrodia camphorata to CCl 4 Protective effect of HepG2 cells induced by acute injury
[0039] 1. Experimental materials and methods.
[0040] HepG2 human liver cancer cells were purchased from the American Type Culture Collection (ATCC), and cells in logarithmic growth phase were used in the experiment. Carbon tetrachloride (Aladdin, Shanghai, China), DMEM medium (Zhongke Maichen, China), DMSO (dimethyl sulfoxide) (AppliChem), premium fetal bovine serum (PAN, South Africa), penicillin- Streptomycin cell culture double antibody (Zhongke Maichen, China), MTS (Promega, USA), silymarin (Huazhong Haiwei, Beijing).
[0041] SpectraMax Plus 384 light absorption microplate reader (Molecular Devices, USA), 3111 cell culture incubator (Thermo, USA), electronic analytical balance (Satorious, Germany).
[0042] CCl 4 Stock configuration: DMSO and CCl 4 Mix 1:1 (v / v), vortex to get 50% CCl 4 stock solution. Add stock solution to DMEM me...
Embodiment 3
[0055] Embodiment 3, Antcin K to CCl 4 Protective effect of acute liver injury caused by
[0056] 1. Experimental materials and methods.
[0057] 4-6 weeks old male ICR mice were purchased from the Experimental Animal Department of Peking University Health Science Center, given 12 hours of light and 12 hours of darkness, constant temperature, normal feed and water, and adapted to a sterile environment for 4 days before the experiment.
[0058] Carbon tetrachloride (Aladdin, Shanghai, China), silymarin (Huazhong Haiwei, Beijing, China), corn oil (Aladdin, Shanghai, China), sodium carboxymethylcellulose (Beijing Chemical Plant, Beijing, China) , deionized water (made by Milli-Q system), paraformaldehyde (Legend Bio, Beijing, China), Tranzol (full gold, Beijing), chloroform, ethanol, isopropanol (Beijing Chemical Plant, Beijing), RNA solution (Quanshijin, Beijing), RNase-free double distilled water, reverse transcription kit (Fermantas, USA), Sybr-green fluorescent dye method q...
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