Antiatherosclerotic sparstolenin B compound and preparation method thereof
A technology of black trilenoid lactone and compound, which is applied in the field of preparation of trilenoid B and the compound, and can solve the problems such as no report on activity, no report on anti-atherosclerosis active component in trilobite, etc.
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Embodiment 1
[0024] The extraction separation method of embodiment 1 black trigonal lactone B (sparstolenin B) compound
[0025] Take 18Kg of dried tubers of Sparganium stoleniferum purchased from Zhejiang, crush them, soak them in 85% ethanol aqueous solution for 2 days, heat and reflux with 85% ethanol aqueous solution to extract three times, combine the three extracts, and concentrate under reduced pressure until there is no alcohol smell , the concentrated solution is diluted with 1 times of water to become a suspension, then extracted with sherwood oil, ethyl acetate, and n-butanol respectively, and the ethyl acetate extract is combined and concentrated to obtain 60.5 grams of extract, and the extract is subjected to a silica gel column (200~ 300 mesh) chromatography, eluted with petroleum ether: ethyl acetate gradient (100:0, 99:1, 98:2, 96:4..., 70:30), and checked by thin layer chromatography, the collection containing black triangular The fractions of lactone B (sparstolenin B) we...
Embodiment 2
[0026] Example 2 Inhibitory effect of trigolide B on arterial smooth muscle cell migration (scratch assay (Chun-ChiLiang, Ann Y Park & Jun-Lin Guan.Nature Protocols, 2007, 2(2), 329))
[0027] Will 1×10 6 Rat arterial smooth muscle cells were seeded in a culture dish, cultured in DMEM medium containing 10% FBS for 24 hours, removed the medium, and then cultured in serum-free DMEM for 24 hours. Scrape a straight line across the cells with a 200 μL pipette tip to create a scratch between the cells and wash the cells twice with DMEM. Add fresh DMEM medium containing 0.5% FBS, and at the same time add different concentrations of trigonolactone B so that the final concentrations are 10, 20 and 50 μg / ml, and set up a blank control. 37°C, 5% CO 2 After continuing to culture under the conditions for 48 hours, take pictures, measure the migration distance of the cells, calculate the average migration distance and perform t test on the data with Prism software. ± 0.035, the blank c...
Embodiment 3
[0028] Example 3 Inhibition of macrophage-mediated LDL oxidation by black trigolide B
[0029] Will 1×10 6 THP-1 cells were seeded in 24-well culture dishes, added RPMI-1640 medium containing 20ng / ml TPA (12-O-tetradecanoylphorbol-13-acetate), at 37°C, 5% CO2 conditions for 72 hours. Discard the culture medium, wash twice with RPMI-1640, add 100 μg / ml human low-density lipoprotein (LDL) and 1 μM CuSO 4 500 μl of fresh culture medium was added, and different concentrations of trigolide B were added at the same time so that the final concentrations were 10, 20 and 50 μg / ml respectively. A blank control group was set up, and 6 replicate wells were set up in each group, and the culture was continued for 4 hours. The cell culture fluid was collected and centrifuged, and 100 μl of the supernatant was taken, analyzed by TBA colorimetry, the OD value was measured at a wavelength of 535 nm by a microplate reader, and the concentration of MDA (malondialdehyde) was calculated to show th...
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