Ficus pumila leaf polysaccharides, preparation method thereof and use thereof in the preparation of anti-complement drugs
A technology of polysaccharides and scallops, applied in drug combinations, pharmaceutical formulas, plant raw materials, etc., can solve problems that have not yet been seen
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Embodiment 1
[0025] Example 1 Preparation of polysaccharides from the leaves of Sapura chinensis BL-PS1, BL-PS3, BL-PS4, BL-PS5 and BL-PS6
[0026] 3Kg of the herb leaves of Ficus praecox were crushed, extracted with 95% ethanol, filtered, the dregs were extracted 3 times with aqueous solution, concentrated, centrifuged, and 4 times the volume of 95% ethanol was added to the supernatant, left to stand, centrifuged to remove the supernatant, precipitated with Redissolve in water, recover under reduced pressure, and remove ethanol; the reconstituted solution is then removed with trichloroacetic acid to remove free protein, centrifuged, and the supernatant is adjusted to neutrality, dialyzed, concentrated, and freeze-dried to obtain crude polysaccharide. 100 g of crude polysaccharide was dissolved in distilled water, centrifuged, and the supernatant was preliminarily separated by a DEAE-cellulose chromatographic column. Elute with distilled water, 0.1, 0.4, 0.8, 1.6 and 2.0 mol / L NaCl solutio...
Embodiment 2
[0033] Example 2 Structural Characterization of Polysaccharides from Fructus Lilii Leaf (BL-PS1, BL-PS3, BL-PS4, BL-PS5 and BL-PS6)
[0034] (1) Determination of molecular weight
[0035] 18-angle laser light scattering gel chromatography system is used to detect molecular weight. The basic principle is that homogeneous polysaccharides pass through gel permeation chromatography to form symmetrical chromatographic peaks, and form light scattering after being irradiated by 18-angle laser light. The light scattering signal is directly related to the molecular weight. . The data is calculated with Astar (version 5.3.1) software and directly gives the molecular weight;
[0036] Experimental method: Accurately weigh 5.0 mg of homogeneous polysaccharide, make it into a 10 mg / ml solution, and pass through a 0.45-micron microporous membrane before sample injection. Chromatographic conditions: flow rate 0.5mg / ml, injection volume 20μL, 0.1% NaCl solution as mobile phase, column temper...
Embodiment 4
[0059] Example 4 Alternative Pathway Complement Inhibition Test
[0060] Serum was taken from healthy adult male volunteers and mixed with VBS-Mg-EGTA buffer (barbital buffer, pH=7.4, containing 5mM Mg 2+and 8mM EGTA) diluted to 1:8, as the complement source of the bypass pathway; rabbit red blood cells stored in 3.8% sodium citrate solution were configured into 0.5% rabbit red blood cells with VBS-Mg-EGTA buffer solution; accurately weighed polysaccharides about 3 mg, add VBS-Mg-EGTA buffer, double-dilute to 8 concentrations, 150 μL of polysaccharide solution of different concentrations and 150 μL of 1:8 complement are pre-incubated at 37°C for 10 minutes, add 200 μL of 0.5% rabbit red blood cells, and incubate at 37°C After bathing in water for 30 minutes, put it into a low-temperature high-speed centrifuge, and centrifuge at 5000 rpm and 4°C for 10 minutes. Take 200 μL supernatant from each tube in a 96-well plate, and measure the absorbance at 405 nm. At the same time, a...
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