Active compound in gastrodiaelata and preparation method and application thereof
A compound, Gastrodia elata technology, applied in the field of medicine, can solve problems such as the lack of a unified theory of aging, and achieve the effect of delaying aging
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Embodiment 1
[0044] 1, the preparation of active compound 1 in Gastrodia elata, concrete steps are as follows:
[0045] (1) Put 1.2kg of dried rhizome of Gastrodia elata in 80% methanol aqueous solution, extract twice at room temperature, each time for 24 hours (shaking); after concentrating by suction filtration, 121 g of extract is obtained.
[0046] (2) Dissolving the obtained extract in water, and extracting the water-soluble matter with ethyl acetate to obtain two components of the water layer and the ethyl acetate layer.
[0047] (3) Separate the water layer by reverse silica gel column chromatography, using methanol: water solvent system as eluent, methanol: water solvent system by volume ratio 15:85, 20:80, 25:75, 30:70 , 35:65, 40:60, 70:30 and 100:0 were eluted in sequence, and 1.8 g of fractions eluted at a volume ratio of 35:65 and 30:70 were taken.
[0048] (4) The fraction obtained in step (3) is separated by forward silica gel column chromatography, dichloromethane: methano...
Embodiment 2
[0058] Active compound 1 prolongs replicative lifespan analysis in embodiment 2 Gastrodia elata
[0059] At present, the biological models used for anti-aging research mainly include yeast, fruit flies, nematodes and mice. In this example, Saccharomyces cerevisiae was selected as the active system for anti-aging research; because yeast is a single-celled eukaryote with a short life cycle, its complete genome data has been obtained, and it is currently a commonly used aging model organism.
[0060] At the same time, resveratrol is used as a positive control, which is a well-known small molecular compound that shows anti-aging effects in various animal models.
[0061] The steps of the analysis method are as follows:
[0062] (1) Take out the K6001 yeast strain from the refrigerator at -30°C, wash it three times with PBS, 5ml each time, and remove the glycerol therein; then add 1ml PBS, pipette to suspend it, and then add it to 5ml liquid medium (1% yeast powder , 2% peptone, ...
Embodiment 3
[0069] Active compound 1 prolongs the sequential lifespan analysis in embodiment 3 Gastrodia elata
[0070] (1) Day -3: Take out the YOM36 yeast strain from the -30°C refrigerator, wash with PBS three times, 5ml each time, and remove the glycerol; then add 1ml of PBS, pipette, dilute to a certain number of times and count with a cell counting plate.
[0071] (2) After the glass petri dish is sterilized, pour 20 mL of sterilized and high-temperature solid medium (1% yeast powder, 2% peptone, 2% glucose, 2% Agar powder), after the culture medium is solidified, add a certain volume of yeast solution according to the dilution factor, so that the number of yeast in each dish is about 200, spread it evenly with a spreader, and put it in a 28°C constant temperature incubator Cultured in medium for 2 days to restore growth ability.
[0072] (3) Day -1: Pick a single yeast colony from the solid medium and inoculate it into liquid medium (0.17% yeast nitrogen source (without amino acid...
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