Preparation method of ganoderan nano-selenium, ganoderan nano-selenium and application thereof
A technology of Ganoderma lucidum polysaccharide and nano-selenium, applied in nanotechnology, nanotechnology, nanomedicine, etc., can solve problems such as lack of research results, and achieve the effects of good storage stability, good pH tolerance and high safety
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Embodiment 1
[0047] 1) Preparation of Ganoderma lucidum polysaccharide nano-selenium solution
[0048]Sodium selenite was prepared into a 50mM solution with first-grade water, ascorbic acid was prepared into a 20mM solution, and Ganoderma lucidum polysaccharide was prepared into a 1mg / mL solution. Mix the sodium selenite solution evenly, add the volume to 10mL with first-grade water, then add 10mL of ascorbic acid solution dropwise, so that the final concentration of Ganoderma lucidum polysaccharide is 25, 50, 100, 200, 300 or 400 μg / ml, and the mixed solution After stirring and reacting for 12 hours at 30°C in the dark, the solution was transferred to an 8000Da dialysis bag, and dialyzed at 4°C in the dark for 72 hours to remove excess ascorbic acid and sodium selenite, and the particle size was measured using a nano-selenium particle size analyzer. The results are as follows figure 1 shown. from figure 1 It can be seen that, compared with the nano-selenium without Ganoderma lucidum pol...
Embodiment 2
[0054] 1) Stability evaluation of Ganoderma lucidum polysaccharide nano-selenium
[0055] The prepared nano-selenium polysaccharides of Ganoderma lucidum were stored at 4°C for 12 weeks, and then stored at different temperatures (4, 25, and 37°C) for 4 weeks. diameter, calculate the average value, the result is as Figure 12 , 13 shown. from Figure 12 It can be seen that the nano-selenium particle size of Ganoderma lucidum polysaccharides has no significant change after being placed at 4°C for 12 weeks, and the storage stability is high; from Figure 13 It can be seen that during the experimental storage period of 4 weeks, the nano-selenium particle size of Ganoderma lucidum polysaccharides placed at 4 °C did not change significantly, while the particle size of Ganoderma lucidum polysaccharide nano-selenium at 25 and 37 °C increased significantly to varying degrees. Large, and the largest change is at 37°C.
[0056] Use hydrochloric acid and sodium hydroxide solution to ...
Embodiment 3
[0058] 1) Toxicity of Ganoderma lucidum polysaccharide nano-selenium to cells
[0059] Mouse liver cancer cell Hepa1c1c7, mouse fibroblast NIH3T3, human liver cancer cell HepG2, human normal liver cell LO2 (5×10 3 per well) inoculated in Corning 96-well plate, at 37°C, 5% (CO 2 Concentration) of CO 2 Cultivate in the incubator for 24 hours; discard the old culture medium, add different concentrations of Ganoderma lucidum polysaccharide nano-selenium (2, 4, 8, 16, 32 μM), add fresh medium to the blank control group, and cultivate for 72 hours; take out the 96-well plate, discard Add 100 μL of serum-free medium containing 0.25 mg / mL MTT (thiazolium blue) to the old culture medium, at 37°C, 5% (CO 2 Concentration) of CO 2 Incubate in the incubator for 2 hours; take out the 96-well plate, discard the MTT (thiazolium blue), add 200 μL of dimethyl sulfoxide, put it in a shaker at 37°C at a speed of 100 rpm / min for 15 minutes, and then use a microplate reader to measure the 550nm ...
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