Prunella vulgaris polysaccharide, and preparation method and purpose thereof
A technology of Prunella vulgaris and polysaccharides, which is applied in the direction of pharmaceutical formulas, medical preparations containing active ingredients, organic active ingredients, etc.
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Embodiment 1
[0024] Embodiment 1. Preparation of homogeneous polysaccharides PW-PS1 and PW-PS2
[0025] The medicinal material of Prunella vulgaris is crushed, extracted by percolation with 95% ethanol at room temperature until it is colorless, and the residue evaporates to remove the alcohol smell. After drying, it is boiled in boiling water for 3 times, adding 4 times the volume of water each time, and the extraction time is 2 hours each time. The water extracts obtained by each extraction and filtration were combined, concentrated to a small volume, and precipitated with 4 times the volume of absolute ethanol. After the precipitate was dissolved in a small amount of water, it was deproteinized with 15% trichloroacetic acid at 4°C. The solution was centrifuged to remove the precipitate, the supernatant was adjusted to neutral with 1mol / L NaOH, and dialyzed against flowing water for 3 days. The solution in the bag was concentrated to an appropriate volume, and freeze-dried to obtain the c...
Embodiment 2
[0028] Example 2. Anti-complement classical pathway test in vitro
[0029] Take 0.1ml of complement (guinea pig serum), add barbiturate buffer solution (BBS) to prepare a 1:5 solution, and double-dilute with BBS to 1:10, 1:20, 1:40, 1:80, 1: 160, 1:320 and 1:640 solutions. Take 1:1000 hemolysin, 0.1ml of each concentration of complement and 2% sheep red blood cells (SRBC) and dissolve them in 0.3ml of BBS, mix well, put them into a low-temperature high-speed centrifuge after 30 minutes in a water bath at 37°C, and put them in a low-temperature high-speed centrifuge at 5000rpm, 4°C. Centrifuge for 10 min under conditions. Take 0.2ml of the supernatant from each tube and place it in a 96-well plate, and measure its absorbance at 405nm. At the same time, a complete hemolysis group (0.1ml 2% SRBC dissolved in 0.5ml triple distilled water) was set up in the experiment. The absorbance of three-distilled water lysed blood vessels was used as the standard of total hemolysis, and th...
Embodiment 3
[0030] Example 3. Anti-complement alternative pathway test in vitro
[0031] Take 0.2ml of complement (human serum), add AP to dilute (barbital buffer, pH=7.4, containing 5mM Mg 2+ , 8mM EGTA) solution was prepared into a 1:5 solution, and double-diluted into 1:10, 1:20, 1:40, 1:80, 1:160, 1:320 and 1:640 solutions. Take 0.15ml of complement of each concentration, 0.15ml of AP diluent and 0.20ml of 0.5% rabbit erythrocytes (RE), mix well, place in a low-temperature high-speed centrifuge after 30 minutes in a 37°C water bath, and centrifuge at 5000rpm and 4°C for 10 minutes. Take 0.2ml of the supernatant from each tube and place it in a 96-well plate, and measure the absorbance at 405nm. At the same time, a complete hemolysis group (0.20ml 0.5% RE dissolved in 0.3ml triple distilled water) was set up in the experiment. The absorbance of three-distilled water lysed blood vessels was used as the standard of total hemolysis, and the hemolysis rate was calculated. Taking the dil...
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