Application of fucoidan in the preparation of medicine and health-care products for preventing and treating neurodegenerative diseases
A fucoidan sulfate, neurodegenerative technology, used in nervous system diseases, neuromuscular system diseases, applications, etc.
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Embodiment 1
[0026] Example 1: Preparation of fucoidan sulfate with fresh sea cucumber as raw material.
[0027] Fresh sea cucumber, gutted, washed, broken into 1cm pieces 3 The left and right pieces are homogenized by adding 5-15 times of water.
[0028] The homogenate was subjected to pepsin (the amount of enzyme was 0.4% of the wet weight of sea cucumber, pH 2.0-2.5, temperature 50°C, time 5h), trypsin (the amount of enzyme was 0.45% of the wet weight of sea cucumber, pH 8.0-8.5, Temperature 50°C, time 5h) enzymolysis, after enzymolysis, boiling to inactivate the enzyme activity, centrifugation (10000r / min, 10min) to take the supernatant.
[0029] The supernatant was precipitated with alcohol, centrifuged (6000r / min, 15min) to obtain the precipitate, then dissolved in distilled water, separated through a DEAE cellulose column, and the absorption peak was detected at 210nm, and the active component was collected (using the activity of nerve cells as an index), and lyophilized.
[0030...
Embodiment 2
[0032] Embodiment 2: Preparation of fucoidan sulfate with kelp as raw material
[0033] Lightly dried kelp is washed, dried and pulverized. Weigh a certain weight of powder (50g), add 5 times the volume (250ml) of absolute ethanol to reflux and extract twice, each time for 2 hours, after centrifugation, place the precipitate on a watch glass to evaporate the ethanol.
[0034] Use 5 times volume of acetone (250ml) to reflux extract the kelp powder twice, each time for 2 hours, centrifuge to get the precipitate and evaporate the acetone to dryness. Add 40 times of water (2000ml) and extract in a 70°C water bath for 5 hours, and filter with gauze to obtain the extract. Add 20 times of hydrochloric acid, pH = 6, and extract at 60°C for 6 hours. Concentrate on a rotary evaporator. Precipitate in 30% ethanol overnight, and centrifuge to retain the supernatant. Precipitate overnight in 60% ethanol: centrifuge to retain the precipitate. Freeze-drying to obtain crude fucoidan. Et...
Embodiment 3
[0035] Example 3: Dedifferentiation of neural precursor cells by fucoidan sulfate.
[0036] Wistar rats at 13.5-14 days pregnant were sacrificed by cervical dislocation.
[0037] Isolate and take out the embryos under aseptic conditions, and separate them in a sterile petri dish under a dissecting microscope to obtain nerve cells. Place them in a carbon dioxide incubator at 37°C and 5% carbon dioxide saturated humidity for 48 hours before performing group experiments.
[0038] After the nerve cells were cultured in the 96-well plate for 48 hours, 5 mg / ml of the fucoidan sulfate prepared in Example 1 were added to make the final concentrations of the fucoidan sulfate respectively 100 μg / ml, 200 μg / ml, and 300 μg / ml, 400μg / ml, a negative control group (blank), and a positive control group for nerve growth factor. Incubate for 48 hours at 37°C, 5% carbon dioxide saturated humidity.
[0039] The analysis results showed that fucoidan sulfate had obvious dedifferentiation effect...
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